活体小鼠中单个红细胞的拉曼光谱分析

Raman Spectroscopic Analysis of Single Red Blood Cells in Vivo Rat

应用拉曼光谱与光镊技术对活体小鼠毛细动脉和静脉血管中的全血以及毛细血管中的红细胞和体外的全血及单个红细胞进行研究;光谱分析显示,可以获得活体血液的拉曼光谱;在动脉毛细管和静脉毛细管中可以观察到清晰的携氧与去氧血液的不同光谱;体内血管中血红细胞的血红蛋白浓度比体外的更高,在动脉毛细管中的血红细胞的携氧态特征峰明显,且具有统计学差异;动脉毛细血管中的pH值比静脉毛细血管中的pH值更高。通过对1604 cm-1特征峰在活体毛细血管中携氧态和去氧态红细胞光谱差异的研究发现,1604 cm-1特征峰的变化与红细胞所处的环境的pH值有关,当环境pH值变大,1604 cm-1特征峰的强度减小;当环境的pH值变小时,1604 cm-1特征峰的强度变大。以上结果表明,在活体中研究血液及单个红细胞的信息变化是可以实现的,同时表明拉曼光谱与光镊技术是一种进行活体中单个细胞研究的强有力的工具。

Raman spectroscopy was used to study red blood cells（RBCs） and blood in vein capillary,in artery capillary in vivo and in vitro with the use of optical tweezers.Results showed that,（1） We could get Raman spectroscopy of blood in vivo;（2） We could clearly observe that different blood spectra in oxygenated and deoxygenated state in artery and vein capillary;（3） By studying Raman spectroscopy of single RBC,we concluded that the hemoglobin concentration in capillary was larger than that in vitro,and oxygenated Raman peak of RBC in artery capillary was more obvious than those in vein capillary;（4） By comparing Raman spectrum in vein with that in artery,we found that that pH value in artery capillary is higher than that in vein capillary,and by studying the change of the band at 1604 cm-1,we found that the band change was in connection with the circumstance pH value around RBCs.These findings make it possible to study conformational changes within single cells in vivo.The experiments also have shown that laser tweezers Raman spectroscopy can provide a strong tool for research in vivo.