407nm光诱导的红细胞荧光光谱随浓度增加红移的机理分析_英文__cropped

407nm光诱导的红细胞荧光光谱随浓度增加红移的机理分析_英文__cropped Vol132 No13 第 32 卷第 3 期光子学报 March 2003 2003 年 3 月 ACTA PHOTONICA SINICA of 407nm Light2induced Erythrocyte Analysis Fluorescence Spectra Red 3Shifted Mechanism with the Concentration Increase 1 ,2 1 ,2 2 2 , 3 3Gao Shumei, Liu Ying, Lu J ian, Ni Xiaowu 1 Department of Physics , Xuzhou Normal University , Xuzhou 221009 2 Department of Applied Physics , N anjing University of Science and Technology , N anjing 210094 Abstract 407nm light excited erythrocyte fluorescence spectra under different concentration in vitro are presented. It is found that there are two prominent and roughly equal fluorescence2emitting regions within the fluorescence spectra . And the fluorescence spectral peaks are apparently red shifted with the increment of the erythrocyte concentration in physiological brine . Furthermore , physical mechanisms of the spectral peaks red2 shifted with the erythrocyte concentrations increasing are analyzed theoretically. Investigation of the fluorescence spectra dependence on the erythrocyte concentrations may makes sense for the light2induced () fluorescence spectra for diagnosis and therapy and the low lever laser therapyLLLTin vivo or in vitro . Key words Fluorescence spectra ; L ED ; Erythrocyte ; Red2shifted ; Fluorophore CL CN Q631 Document Code A 0 Introduction fluorescence spectra are measured in vitro as an exciting The light2induced fluorescence spectra for 1 ,2 () ( ) diagnostics and therapy L IFSTof biological tissues2emitting diode L ED . light of lightThe spectra red2 have been well set up for several decades. Both non2 and the erythrocyte shifted physical mechanism with imaging fluorescence in vivo oncological applications of concentration increase is analyzed theoretically. L IFST have been proved to be helpful for detection of 1 Materials and Methods early2stage lesions when conventional diagnostic tools do not provide needed urgency and accuracy. On the other The experimental set is a WGD28 grating () hand , the low level laser therapies LLLTin vitro and in spectrometer that made in Tianjing Gangdong Corporation. vivo have also been investigated for several decades , and An R928 photomultiplier , made by Hamamatsu Corp . in 3 ,4 achieved significant curative effect. J apan , placed the left exit slit of the spectrometer in order However , when diagnosing cancer tissues by induced to increase the sensitivity of the measurement of fluorescence , washing the biological slice is often fluorescence intensity. The Spectrometer collecting pace2 necessary , and it is certain that there are residual some interval and scanning range are kept at 0 . 5 nm and 200 blood cells on the slice concentration. Handling one′s nm, 850 nm in experiments , respectively. All spectra cases with low lever laser irradiation therapy in vivo , the are displayed at the range of 350 nm to 750 nm laser and liquid medicament —the brine , dextrose , and so thereinafter , the fluorescence beyond the range has not on. The investigation of light 2blood cells interaction plays been found in our experiments. The exciting light is 407 an important role in many diseases treats with LLL IT and (Δλ) nm L ED ?16 nm, which power is 5 mW. 1Π2 L IFST. In addition , researches had indicated that blood Blood from healthy small white mice is separated by 5 fluorescence mainly derived from erythrocyteand light centrifugation with 1600rΠmin , which is 24 cm in induced erythrocyte fluorescence spectral characteristics diameter. And then the erythrocyte is gained after under different concentrations are distinctly different . throwing out above limpid liquid and diluted into various paper , 407 nm light2excited erythrocyte In this concentrations with physiological brine for measuring their 3 fluorescence . Supported by The Education Department′s Scientif ic Foundation of Jiangsu Province ( No. Research 02 KJB140006) and The ′s Key Foundation ( No.University 2 Results and Discussion 01AXL002) 3 3 407 nm L ED2excited erythrocyte fluorescence spectra Tel :025 4315075 Correspondence Author Ema il :nx w @ma il . njust. edu. cn different concentration are shown in Fig. 1 . at Received date :2002 05 08 p s Apparently , there are two prominent fluorescence regions ()( ) ( )4 E= 6 - CΠr+ 6 6 - DΠr n pn s ni p s i within the fluorescence spectra , the first prominent Where p is 2,12 , s is 4,12 , i and n are all positive maxima are about located between 600nm and 650 nm integer , B , Cand Dare constants depend on b p s when the erythrocyte concentration is higher , the second temperature , native and extra macromolecules or group s primary fluorescence peaks appear on the range of 428nm state in a experiment , which are undetermined constants to 513 nm while the concentration is lower , which are ( ) by experiment . In Eq. 4, the first item describes the roughly equal . Furthermore , the two primary fluorescence electronic levels of the fluorophores in the case of no extra peaks are significantly red shifted with the increment of disturbance , where rand rhave been quantized. The n ni erythrocyte concentration. In addition , there are two small later expresses the repellent energies from other peaks respectively located 556nm and 694 nm in the molecules. spectra , but they are not almost changed with the While the fluorophore in ground electronic state erythrocyte concentration. The first fluorescence peaks ( ) suppose to be n absorb energy at an appropriate obviously derive from the 407nm exciting light . The wavelength to move to electronic excitation to the singlet broken lines direct the peak wavelength in Fig. 1 . state and the upper vibrational level . And then the fluorophores lose their excess energy and move to a lower ( vibratonal level in the excited electronic state suppose to ) be m. Lastly , in the excited electronic state , they lose (Δ ) ( γ) their energy Ein the form of light hto regain ground state . Therefore , the energy moving is - p - p Δ ( ) E = E- E= - 6 Cr-r- m n p m n p - s ()( )5 6 6 Dr- r- s s mi ni Fig. 1 407 nm light induced erythrocyte fluorescence spectra s i Δ γ λIt is known : E = h= hcΠ. The electronic transition ( ) energy from the electronic excited state mto the ground 2shifted spectral peaks redThe fluorescence ( ) electronic state nin the case of no extra disturbance is phenomena with the concentration increase may results - p - p Δ ( E= 6 Cr- ) ( )()r from repellent forces fields from other macromolecules or r> r 6 0 p n mm n p group s. Considering synthetically interactional potential Disturbing energies from other molecules is - s - s and electron2 energy among electrons , molecules , Δ ( ) ()E = 6 6 Dr- 7 r s mi nis i molecules , and so on , total potential energy of the ()As a result of r < r , Eq. 5 can be rewritten by mi ni 6fluorophore in an erythrocyte is ()Δ Δ Δ 8 E =E- E 0 6 a ( ) ( )()U r= 6 6 - A Πr 1 i ai increase results in Obviously , the concentration rni i a = 2 Where ris distance between the fluorophore researched i Δ Δ E decrease , or E increase . shortened , and then γ λand another one i , A is a constant depend on Therefore , decreased and enlarged. Consequently , it is a explained that the fluorescence peaks are red2 shifted temperature , native and extra macromolecules or group s ( with the increment of the erythrocyte concentration see in ( ) state. U r is supposed potential energy of the ) Fig. 1. fluorophore , where r is distance between an electron on the fluorophore and center of the fluorophore . 3 Concl usion Apparently , r is far larger than r. Therefore , the i A , B , Cand Dmentioned above depend on a b p s Hamilton2functor in Schr ? dinger equation may be ^ 2 2 -temperature , native and extra macromolecules or group s ()μ) ( ) ( )( 2 H = - 2/ A+ U r+ U r i h state. When these conditions are altered , for instance , or 6 6 some diseases , the A , B , Cand Dwill be changed a b p s ^ A μ 2 2 b a( ) ()H = - A+ 6 - B Πr+ 6 6 3 b 2a-b = 1 i a = 2 h rand result in excited different fluorescence spectra . Some i diseases will be diagnosed. Therefore , investigating why So main quantum electronic levels on the fluorophore may be given by , approximatively blood fluorescence spectra depend on their concentration Gao Shumei ,et al . Analysis of 407nm light2induced erythrocyte fluorescence spectra red 3 期 263 shifted mechanism with the concentration increase Fluorescence Spectroscopy from Native Cancerous and Normal may make sense for fluorescence spectra diagnostic Tissue. I EEE J ournal of Quantum Electronics , 1984 , Qe220 techniques and LLL IT in vivo or in vitro . In addition , the () 12: 507,1511molecule with many atoms has very completed energy Pinheiro A. Low2Level Laser Therapy is an Important Tool to 3 structure , in which the number of the energy levels is Treat Disorders in the Maxillofacial Region. J Clinical L aser Med more and more with the number of the atoms. In () & S urg , 1998 , 16 4: 2232226 particular , a biological macromolecule has a thousand 4 Zhu Ping. Low lever laser intravascular irradiaton therapy. 21 different electronic transitions on each cm. Thus , the Nanning : Guanxi Scienc And Technology Press , 1999. 4,15 spectra gained in this article are continuous. 5 Gao Shumei , Luo Xiaosen , Lan Xiufeng , et al . Analyzing of L ED2 induced Blood Fluorescent Spectra at about 570nm. References ( ) ()Chinese J ournal of L asers B , 2002 ,11 4 Li Guoqi , Wang Hemu , Li Anzi . Molecular Biological Physics. 6 1 Profio A E , Ooiron D R , Samaik J . Fluorometer for endoscopic Beijing : Higher Education Press , 1992. 55,81 () diagnosis of tumors. Medical Physics , 1984 ,11 4: 516,520 2 Alfano R R , Tata D B , Cordero J , et al . Laser Induced 407 nm 光诱导的红细胞荧光光谱随浓度 增加红移的机理分析 高淑梅刘莹陆健倪晓武 ()1 徐州师范大学物理系 ,江苏 221009 () 2 南京理工大学应用物理系 ,江苏 210094 收稿日期 :2002 05 08 摘 要 用 407 nm 光激发不同浓度离体红细胞可以产生较强的荧光光谱 ,进一步研究发现这些荧光光谱中 存在两个明显的 、频谱宽度大致相等的荧光发射区 ,而且在生理盐水中随着红细胞浓度的增加 ,其发射的荧 光光谱的谱峰值位置向长波方向移动 ,即出现“红移”现象. 从理论上对这种“红移”现象的产生机理进行了分 析 ,明这种现象是由于浓度变化对荧光团电子能级产生的微扰发生变化所致. 该研究结果对现代医学中的 荧光光谱诊断技术和低功率激光照射疗法都有一定的参考价值. 关键词 荧光光谱 ,L ED ;红细胞 ;红移 ;荧光团 Gao Shumei is an associate professor of physics in Department of Physics , Xuzhou Normal University and a doctoral graduate in Biomedical Engineering in Department of Applied Physics , Nanjing University of Science and Technology. In recent years her recearch has been concerned with applications of lasers in medicine and multi2holography , particularly for the investigation of the mechanisms of light2tissue interaction