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细胞工程 动物组织和细胞培养

2017-10-12 50页 doc 593KB 8阅读

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细胞工程 动物组织和细胞培养细胞工程 动物组织和细胞培养 第3章 动物细胞和组织培养工程 第一节 植物组织和细胞培养的基础知识 一 动物细胞培养的基本条件 无菌条件:净化工作室,风淋室,传递窗,高效过滤器, 洁净层流罩,生物安全柜,超净工作台,紫外 灯,电热干燥箱,高压灭菌器,抗生素 细胞生长条件:纯水蒸馏器,纯水仪,培养板,培养瓶, CO2培养箱,培养基,血清 细胞检测条件:倒置显微镜,酶标仪 ,微孔板震荡器,高 速离心机,移液器 细胞保存条件:液氮罐 ItAAAASA Close to the desired solution...
细胞工程 动物组织和细胞培养
细胞 动物组织和细胞培养 第3章 动物细胞和组织培养工程 第一节 植物组织和细胞培养的基础知识 一 动物细胞培养的基本条件 无菌条件:净化工作室,风淋室,传递窗,高效过滤器, 洁净层流罩,生物安全柜,超净工作台,紫外 灯,电热干燥箱,高压灭菌器,抗生素 细胞生长条件:纯水蒸馏器,纯水仪,培养板,培养瓶, CO2培养箱,培养基,血清 细胞检测条件:倒置显微镜,酶标仪 ,微孔板震荡器,高 速离心机,移液器 细胞保存条件:液氮罐 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 实验室安全:生物实验室安全,P1,P2,P3实验室安全 (一) 无菌条件 ? 净化工作室,风淋室,传递窗,高效过 滤器,洁净层流罩,生物安全柜,超净 工作台,紫外灯,电热干燥箱,高压灭 菌器,抗生素. ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 1 净化工作室 ------ 我国药品生产洁净室(区)空气洁净度标准 ------- 尘粒最大允许数 尘粒最大允许数/立方米 洁净度级 别 ?0.5um ?5um 浮游菌/立方 沉降菌/皿 100 3,500 0 5 1 350,000 2,000 100 3 10,000 100,000 3,500,000 20,000 500 10 300,000 60,000 NA 1510,500,000 2 风淋室: ?风淋室是生物洁净室 的理想配套设备,用于 吹除进入洁净厂房的人、 物表面附着的尘埃 ?是进行人身、物料净 化和防止室外空气侵入 洁净区的有效设备 ?风淋室可以配合加热 器,冬天可以加热,温 度可调,一般控制温度 在30-35?较为适宜。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 3 传递窗: 该装置是一种洁净室 的辅助设备,主要用于 洁净区与洁净区,洁净 区与非洁净区之间小件 物品的传递,以减少洁 净室的开门次数,把对 洁净区的污染降低到最 低程度。 4 高效过滤器: ?高效过滤器是用超细玻璃纤维 纸作滤料,胶板纸作分隔板,可 与铝合金框、木框及镀锌钢板框 组合而成。 ?特点:具有低阻高效、轻质、 容尘量大等特点, ?适用于常温常压常湿条件下环 境空气的净化. ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 5 洁净层流罩: 层流罩是可提供局部高洁净 环境的空气净化设备。洁净 层流罩是将空气经风机以一 定的风压通过高效空气过滤 器后,使洁净空气呈垂直气 流送入工作区,从而保证了 工作区内达到工艺所需的高 洁净度。 6 超净工作台 超净工作台的工作 原理是利用鼓风机 驱动空气通过高效 滤器除去空气中的 尘埃颗粒,使空气 得到净化。净化空 气徐徐通过工作台 面,使工作台内构 成无菌环境。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 7 生物安全柜: 一种既能使操作造成无菌、 无尘的局部空气环境,保证检 验、检测不受污染,又能保证 被研究的对象(如病菌、细菌 等)不外溢,保护周围环境及 操作人员的安全隔离设备。 该设备可广泛应用于低、 中等危险度的临床细菌学,病 毒学、微生物学、组织培养、 生物学及生命科学的研究,加 工、检验部门。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ? 如何除菌, ? 灭菌 8 紫外灯 :紫外线消毒。主要用于培养室空气、 操作台、塑料培养皿和培养板等表面消毒 。 9 电热干燥箱:干热消毒(160 ? ,2小时)。主要用干玻璃 器皿消毒 。 10蔡氏( Zeiss)滤器:过滤除菌,滤器中的滤膜 一般用0.2,0.3μm孔径的微 孔滤膜。凡在高温或 射线下易发生变性或失去功能的物质,如人工合 成培养液、血清、消化 用胰酶等都须用滤器过滤 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 除菌。 滤 器 大容量高压灭菌器 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept (二) 细胞生长条件 ? 纯水蒸馏器,纯水仪,培养板,培养瓶, CO2培养箱,培养基,血清 1自动双重纯水蒸馏器 2 纯水仪 精 滤 、 活 性 碳 吸 附 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 反渗透 3 标准型细胞培养板 (Tissue Culture Plates) ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 6孔,12孔,24孔,48孔,96孔 4 培养瓶 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 5 CO2培养箱 CO2培养箱主要由箱体结构 件、温度控制系统和CO2浓度 控制系统三部分组成。 CO2培养箱设定的条件为37 ? ,5,CO2 。 再现活体环境对于温度、PH值 CO2浓度、氧分压(O2浓度)、相 对湿度等细胞生长关键的因素, 6 移液器 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept (三) 细胞检测条件: ? 倒置显微镜,酶标仪 ,微孔板震荡器, 高速离心机,移液器 15 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 用于微生物、细胞、细菌、组织 培养、悬浮体、沉淀物等 的观察,并可将此过程中的任一形态拍摄下来。 酶标仪 3 微孔板震荡器 2 主要用于酶标板、细胞培 通过自动酶标仪计数来检测活细胞 养板(如6孔板、12孔板、 数量。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 24孔板、48孔、96 孔板 等)等溶液的混匀或细胞 的混匀。 (四) 细胞保存条件: ? 液氮罐 -196 ? ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept (五) 实验室安全: ? 生物实验室安全,P1,P2,P3实验室安全 实验室安全 层流净化细胞培养室管理规则 一级生物安全防护实验室 二级生物安全防护实验室 三级生物安全防护实验室 四级生物安全防护实验室 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 层流净化细胞培养室管理规则 层流净化细胞培养室的总体要 局部小环境空气 求:经济、有序、高效、安全。 达到洁净度等级 初步制定相关如下: 实验室安全级别 一级生物安全防护实验室 实验室结构和设施、安全操作规程、安全设备适用于 对健康成年人已知无致病作用的微生物,如用于教学 的普通微生物实验室等。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 二级生物安全防护实验室 实验室结构和设施、安全操作规程、 安全设备适用于对人或环境具有中等 潜在危害的微生物。 二级生物安全防护实验室 1 可能产生致病微生物气溶胶或出现溅出的操作均应在生物安全柜(?级生物 安全柜为宜)或其他物理抑制设备中进行,并使用个体防护设备。 2 处理高浓度或大容量感染性材料均必须在生物安全柜(?级生物安全柜为宜) 或其他物理抑制设备中进行,并使用个体防护设备。 上述材料的离心操作如果使用密封的离心机转子或安全离心杯,且它们只在生物 安全柜中开闭和装载感染性材料,则可在实验室中进行。 3 当微生物的操作不可能在生物安全柜内进行而必须采取外部操作时,为防 止感染性材料溅出或雾化危害,必须使用面部保护装置(护目镜、面罩、个体呼吸 保护用品或其他防溅出保护设备)。 4 在实验室中应穿着工作服或罩衫等防护服。离开实验室时,防护服必须脱 下并留在实验室内。不得穿着外出,更不能携带回家。用过的工作服应先在实验 室中消毒,然后统一洗涤或丢弃。 5 当手可能接触感染材料、污染的表面或设备时应戴手套。如可能发生感染 性材料的溢出或溅出,宜戴两副手套。不得戴着手套离开实验室。工作完全结束 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 后方可除去手套。一次性手套不得清洗和再次使用。 三级生物安全防护实验室 实验室结构和设施、安全操作规程、安全设备适用于 主要通过呼吸途径使人传染上严重的甚至是致死疾病 的致病微生物及其毒素,通常已有预防传染的疫苗。 例如:艾滋病病毒的研究(血清学实验除外)应在三级 生物安全防护实验室中进行。 例如:重症SARS病人传染性 较强,这与病人产生的大 量呼吸道飞沫有关。ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 三级生物安全防护实验室 1 实验室中必须安装?级或?级以上生物安全柜。 2 所有涉及感染性材料的操作应在生物安全柜中进行。当这类操作不得 不在生物安全柜外进行时,必须采用个体防护与使用物理抑制设备的综合防 护措施。 3 在进行感染性组织培养、有可能产生感染性气溶胶的操作时,必须使 用个体防护设备。 4 当不能安全有效地将气溶胶限定在一定范围内时,应使用呼吸保护装 置。 5 工作人员在进入实验室工作区前,应在专用的更衣室(或缓冲间)穿着 背开式工作服或其他防护服。工作完毕必须脱下工作服,不得穿工作服离开 实验室。可再次使用的工作服必须先消毒后清洗。 6 工作时必须戴手套(两副为宜)。一次性手套必须先消毒后丢弃。 7 在实验室中必须配备有效的消毒剂、眼部清洗剂或生理盐水,且易于 取用。可配备应急药品。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 四级生物安全防护实验室 实验室结构和设施、安全操作规程、安全设备适用于对人 体具有高度的危险性,通过气溶胶途径传播或传播途径不 明,目前尚无有效的疫苗或治疗方法的致病微生物及其毒 素。与上述情况类似的不明微生物,也必须在四级生物安 全防护实验室中进行。待有充分数据后再决定此种微生物 或毒素应在四级还是在较低级别的实验室中处理。 1 在实验室中所有感染性材料的操作都必须在?级生物安全柜 中进行。如果工作人员穿着整体的由生命维持系统供气的正压工作 服,则相关操作可在?级生物安全柜中进行。 2 所有工作人员进入实验室时都必须换上全套实验室服装,包 括内衣、内裤、衬衣或连衫裤、鞋和手套等。所有这些实验室保护 服在淋浴和离开实验室前均必须在更衣室内脱下。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept (六) 培养细胞生长的条件 1 细胞的营养需要 2 细胞的生存环境 温度: 37 ?,O2 CO2: 5%,CO2 +H2O ? ?H2CO3 ? ?H+ + HCO3- pH: 7.2-7.4 3 无污染 4 无毒 1 细胞培养用液的配制 1)水:新鲜配置的三蒸水或去离子水 2)平衡盐溶液:无Ca2+、Mg2+的缓冲液,维持一定渗 透压和pH PBS:NaCl 8.0 g KCl 0.2 g Na2HPO4.H2O 1.56 g 0.20KH2PO4 加水至1000 ml ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 3)消化液: 胰蛋白酶作用于与赖氨酸或精氨酸相连接的肽健,除去细 ? 胞间粘蛋白及糖蛋白,影响细胞骨架,从而使细胞分离。 胰蛋白酶是一种黄白色粉末,用无Ca2+、Mg2+的PBS缓冲液配制, ? 常用的胰蛋白酶液浓度是0.25, 。用滤器过滤除菌。 胰蛋白酶液消化时间:2-10分钟。? 4) 培 养 基 : 培养基(培养液)是维持体外细胞生存和生长的溶液,分天然 ? 培养基和合成培养基。 ?天然培养基: ? 天然培养基有血清、血浆、和组织提取液(如鸡胚和牛胚浸 ? 液)。 优点:营养成分丰富,培养效果好 ? 缺点:来源受限 ? 成分复杂,影响对某些实验产物的提取和实验结果的分析。 ? 易发生支原体污染? ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ?合成培养基: 合成培养基是根据细胞生存所需物质的种类和数量,用人 工方法模拟合成的。目前已设计出许多种培养基,如TC199、 MEM、RPMI-1640、 DMEM等。 合成培养基主要成分是氨基酸、维生素、碳水化合物、无 机盐和其它一些辅助物质。 优点:标准化生产,组分和含量相对固定。成本低 。 缺点: 缺少某些成分,不能完全满足体外细胞生长需要。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept 人工合成培养基只能维持细胞生存,要想使细胞生长和繁 殖,还需补充一定量的天然培养基(如血清)。 血清中含有: ?多种蛋白质(白蛋白、球蛋白、铁蛋白等) ; ?多种金属离子; ?激素; ?促贴附物质,如纤粘蛋白、冷析球蛋白、胶原等; ?各种生长因子; ?转移蛋白; ?不明成分。 ? 一般说来,含5,小牛血清的培养基对大多数细胞可以维 持细胞不死,但支持细胞生长一般需加10,血清( 血清中不仅存在促细胞生长因子,同对也存在细胞生长 ? 抑制因子或毒性因子, ? 因此在含血清培养基中培养的细胞所反映的生物学特性 是细胞和复杂血清因子的综合反应。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ? ?无血清培养基:在基础培养基中补充各种必需因 子,如激素、生长因子 、结合蛋白 、贴壁和扩展 因子 等。 ? 无血清培养基由基础培养基和替代血清的补充成 分组成。 无血清细胞培养基优点保证了实验结果的准确性、可重复 ? 性和稳定性,减少了细胞污染,简化了提纯和鉴定各种细 胞产物的程序。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ? 血清质量好坏是实验成败的关键。 ? 常用血清有胎牛血洁、新生牛血清、小牛血清、 兔血清、马血清等,其中以胎牛血清质量最好。 ? 优质血清的标准:透明,淡黄色,无沉淀物,无 细菌、支原体、病毒污染。 ? 血清的灭活(消除补体活性):56 ? ,30 分钟 ? 血清的消毒:过滤除菌 补体是热敏性的,是机体免疫防御系统的重要组成部分, 其主要功能是抗感染,补体虽然具备杀死病菌的作用,但 有时也伤害好细胞,引起炎症的作用。 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept ? 抗菌素的使用: 在培养液配制后,培养液内常加适量抗菌素,以抑制可 ? 能存在的细菌的生长。 通常是青霉素和链霉素联合使用。培养基内青霉素、 ? 链霉素最终使用浓度为每毫升100单位。 庆大霉素:每毫升100单位——方便、广谱、稳定。? ?完全培养基的组成 ? 基础培养基 80,一95, ? 血清 5,一20, ? 碳酸氢钠 2.0 g/L ? 青、链霉素 各100单位,毫升 ItAAAASA Close to the desired solution was prepared, and then use the reference material (or another material standard) to determine its exact concentration. This determines its exact concentration is called calibration. For example, for the preparation of 0.1mol • L-1HCl standard solution, first with a certain amount of concentrated HCl diluted with water, mixed concentration is about 0.1mol • L-1 in dilute solution, and then use the anhydrous Na2CO3 benchmark solution titration accurately weigh the material until both quantitative response fully, and then consumed in the titration of aqueous solution of HCl volume and quality of anhydrous Na2CO3, calculate the exact concentration of the HCl solution. Most accurate concentrations of the standard solution are determined by calibration of the method. Constant component in the determination of standard solution concentration range of 0.01 mol • L-1-1 mol • L-1, typically according to the components to be tested to select the height of the size of standard solution concentration. In order to improve the accuracy of calibration, calibration should pay attention to the following points: ? determination of calibration should be parallel 3-4, repeated at least three times, and called for determination of the relative deviation is less than 0.2%. ? in order to reduce measurement error, weighing the baseline amount of substance should not be too few, weighing at least 0.2G above; same titration at the end consumption of standard solutions or too small in size, the best is 20mL. C preparation of calibration solutions and use liquor, such as glassware, such as volumetric flasks and pipettes, where necessary, corrected volume, and considering the effect of temperature. D calibration good of standardOperated by stock solution was prepared solution, in principle only be diluted once, if necessary, dilute the secondary. Dilution of too many accumulated error is too large, affect the accuracy of the results. 2. calibration cannot be directly made accurate concentrations of the standard solution, first solution was prepared, and then select the base material calibration. Do the titration with acid and alkali solution, it is usually prepared with about 0.1mol • The concentration of L-1. Of solid acid and alkali solution prepared from original, generally only accurate to 1~2 significant figures, it can be graduated cylinders measuring liquids or solid reagents weigh in the scales, add the solvent (water), graduated cylinder or measuring cup amounts. But in the whole process of calibration solution, everything is very strict and accurate. Weighing standard substances requires the use of analytical balance, accurate to four digits after the decimal point. Volume calibration solution, if it were to participate in the concentration are calculated using volumetric flasks, pipettes, burets accurate operation, not sloppy. (B) the general solution preparation and preservation methods of 1:1 (or 1+1), 1:2 (1+2) volume is expressed as concentration. For example, 1:1 solution of H2SO4, the original concentrated H2SO4 volume 1 volume, mix with 1 volume of water. And as 1:3 HCl, the original volume 1 volume hydrochloric acid and three volumes of water and mix. Preparation of solution, according to the requirements on the accuracy of solution concentration should be determined on the balance at the level of the weighing; record should be written to several significant figures; the prepared solution choose what kind of vessel. That accurate, it should be very strict tolerances can be less stringent. The "quantity" concept
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