光学学报, 2009, 29 (10): 2854, 网络出版: 2009-10-19   

单细胞拉曼光谱分析地中海贫血红细胞的氧合态和去氧态

Single-Cell Raman Spectral Analysis of Oxygenated and Deoxygenated Thalassemia Erythrocytes
作者单位
1 广西科学院生物物理实验室,广西 南宁 530003
2 广西师范大学 物理科学与技术学院,广西 桂林 541004
3 中国人民解放军第303医院,广西 南宁 530022
4 美国东卡罗莱纳大学物理系,Greenville,NC 27858-4353,USA
摘要
应用光镊拉曼光谱系统俘获单个红细胞并收集其拉曼光谱,选择I1638/I1547比值为区分红细胞氧合及去氧态指标,比较分析地中海贫血HbH-CS和健康个体单个红细胞的氧循环过程的氧合和去氧能力及去氧态群体红细胞。结果发现,在氧循环的去氧态中,地中海贫血HbH-CS 的I1638/I1547显著高于正常对照;氧合态的I1638/I1547也略高于正常对照。群体统计中,若以小于正常对照I1638/I1547的平均值为去氧态基准,则正常对照46%的红细胞达到去氧态,而地中海贫血HbH-CS仅有15 %,表明地中海贫血HbH-CS红细胞的氧合能力较强,但去氧能力显著低于正常对照,而且地中海贫血HbH-CS红细胞个体差异较大。该研究在单细胞水平上,揭示了地中海贫血HbH-CS红细胞氧合、去氧能力以及细胞个体差异性,以评价其携氧功能,为诊断治疗提供一定的理论依据。
Abstract
A laser tweezers Raman spectroscopy (LTRS) system was used to investigate the capacity of carrying oxygen in thalassemia erythrocytes. Single erythrocyte from patients with thalassemia HbH-CS and normal donors were trapped,and the Raman scattering signal of trapped cell was collected. The intensity ratio of band 1638 cm-1 to band 1547 cm-1 (I1638/I1547) was selected to distinguish oxygenated erythrocytes from deoxygenated ones. By comparing the oxygenation cycle of optically trapped single red blood cells and the deoxygenated cells population from the two kinds of specimen,and analyzing the oxygenated and deoxygenated states,it is found that,in the oxygenation cycle,the ratio I1638/I1547 of thalassemia HbH-CS was much higher than that of normal control in deoxygenated state and slightly higher than the normal control in oxygenated state. In the population statistic,46% of the red blood cells from the normal control reach oxygenated state,but thalassemia HbH-CS only 15%,if the base is set by less than the average of I1638/I1547 from normal control cells. The result shows that the red blood cells of thalassemia HbH-CS are easy to oxygenate,but difficult to deoxygenate. The individual difference between thalassemia HbH-CS erythrocytes is obvious. This study provides the spectral vision on the oxygenation of thalassemia HbH-CS erythrocytes and the laboratorial referrences for the diagnosis and treatment.
参考文献

[1] B.R. Wood,L. Hammer,L. Davis et al.. Raman microspectroscopy and imaging provides insights into heme aggregation and denaturation within human erythrocytes [J]. J. Biomed. Opt.,2005,10(1):014005

[2] 姚成灿,李校坤,黄耀熊. 温度对单个活态人红细胞携氧能力的即时影响[J]. 光谱学与光谱分析,2005,25(4):613-616

    Yao Chengcan,Li Xiaokun,Huang Yaoxiong. Instant effect of temperature on the oxygen carrying capacity of single living intact red blood cell [J]. Spectroscopy and Spectral Analysis,2005,25(4):613-616

[3] T. G. Spiro,T. C. Streakas. Resonance Raman spectra of heme proteins. Effects of oxidation and spin state [J]. J. Am. Chem. Soc,1974,96(2):338-345

[4] H. Brunner,A. Mayer,H. Sussner. Resonance Raman scattering on the haem group of oxy- and deoxyhaemoglobin [J]. J. Mol. Biol.,1972,70:153-156

[5] X. Zhao,T. G. Spiro. Ultraviolet resonance Raman spectroscopy of hemoglobin with 200 and 212 nm excitation:H-bonds of tyrosines and pralines [J]. J. Raman Spectrosc.,1998,29:49-55

[6] 沈高山,谷怀民,闫天秀 等. 亚硝酸钠和氧合血红蛋白反应的拉曼光谱[J]. 中国激光,2008,35(9):1432-1436

    Shen Gaoshan,Gu Huaimin,Yan Tianxiu et al.. Micro-Raman spectroscopy of the interaction between sodium nitrite and oxyhemoglobin[J]. Chinese J. Lasers.,2008,35(9):1432-1436

[7] B. R. Wood,B. Tait,D. McNaughton. Micro-Raman characterisation of the R to T state transition of haemoglobin within a single living erythrocyte [J]. Biochim. Biophys. Acta,2001,1539:58-70

[8] B. R. Wood,D. McNaughton. Micro-Raman Characterization of High- and Low-Spin Heme Moieties within Single Living Erythrocytes [J]. Biopolymers,2002,67:259-262

[9] B. R. Wood,P. Caspers,G. J. Puppels et al.. Resonance Raman spectroscopy of red blood cells using near-infrared laser excitation [J]. Anal. Bioanal. Chem.,2007,387:1691-1703

[10] K. Ramser,E. J. Bjerneld,C. Fant et al.. Raman imaging and spectroscopy of single functional erythrocytes:a feasibility study [C]. SPIE,2002,4614:20-27

[11] K. Ramser,K. Logg,J. Enger et al.. Resonance Raman study of the oxygenation cycle of optically trapped single red blood cells in a microfluidic system [C]. SPIE,2004,5514:560-568

[12] A. C. De Luca,G. Rusciano,R. Ciancia et al.. Spectroscopical and mechanical characterization of normal and thalassemic red blood cells by Ramantweezers [J]. Opt. Express,2008,16(11):7943-7957

[13] 张俊武,龙桂芳. 血红蛋白与血红蛋白病[M]. 南宁:广西科学技术出版社,2003

    Zhang Junwu,Long Guifang. Hemoglobin & Hemoglobinopathies[M] . Nanning:Guangxi Science & Technology Publishing,House,2003

[14] S. L. Schrier,A. Bunyaratvej,A. Khuhapinants et al.. The unusual pathobiology of hemoglobin Constant Spring red blood cells [J]. Blood,1997,89(5):1762-1769

[15] M. Abe,T. Kitagawa,Y. Kyogoku. Resonance Raman spectra of octaethylporphyrinato-Ni(Ⅱ) and mesodeuterated and 15 N substituted derivatives. Ⅱ. A normal coordinate analysis [J]. J. Chem. Phys. 1978,69:4526-4531

[16] B. R. Wood,D. McNaughton. Raman excitation wavelength investigation of single red blood cell in vivo [J]. J. Raman Spectrosc.,2002,33:517-523

陈秀丽, 王桂文, 尹晓林, 刘军贤, 姚辉璐, 黄庶识, 黎永青. 单细胞拉曼光谱分析地中海贫血红细胞的氧合态和去氧态[J]. 光学学报, 2009, 29(10): 2854. Chen Xiuli, Wang Guiwen, Yin Xiaolin, Liu Junxian, Yao Huilu, Huang Shushi, Li Yongqing. Single-Cell Raman Spectral Analysis of Oxygenated and Deoxygenated Thalassemia Erythrocytes[J]. Acta Optica Sinica, 2009, 29(10): 2854.

本文已被 6 篇论文引用
被引统计数据来源于中国光学期刊网
引用该论文: TXT   |   EndNote

相关论文

加载中...

关于本站 Cookie 的使用提示

中国光学期刊网使用基于 cookie 的技术来更好地为您提供各项服务,点击此处了解我们的隐私策略。 如您需继续使用本网站,请您授权我们使用本地 cookie 来保存部分信息。
全站搜索
您最值得信赖的光电行业旗舰网络服务平台!