Confocal Raman microscopy on single living young and old erythrocytes.
Raman confocal microscopy, including the techniques of point Raman spectra, line mapping, 2D mapping, and time-dependent spectrum monitoring performed with 514.5 nm excitation light, was used in a comparative study on the distribution and oxidation states of hemoglobin (Hb) in young and old mature erythrocytes. It is demonstrated that in contrast to the homogeneous distribution of the Hb in young cells, there are more Hb distribution around the cell membrane in old erythrocyte. The proteins exhibit some extent of aggregation and conformational change, present less ability of oxidation, and lower oxygenation speed than the Hb in young erythrocytes. Our results also provide the first direct evidence of some intermediate oxygenated states of Hb between the two fully oxygenated (R) and deoxygenated (T) states in living erythrocyte, and give detail information about the conformational change of the intracellular Hb with time during the reoxygenation process.
Kang
Li-Li
LL
Department of Biomedical Engineering, Ji Nan University, Guangzhou, China.
Huang
Yao-Xiong
YX
Liu
Wen-Jing
WJ
Zheng
Xin-Jing
XJ
Wu
Zheng-Jie
ZJ
Luo
Man
M
D004912:Erythrocytes / Q000737:chemistry* / Q000166:cytology* / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008853:Microscopy / Q000379:methods*; D010084:Oxidation-Reduction; D013059:Spectrum Analysis, Raman / Q000379:methods*
D006454:Hemoglobins