Continuous intact cell detection and viability determination by CE with dual-wavelength detection.
We introduce here a method for continuous intact cell detection and viability determination of individual trypan blue stained cells by CE with ultraviolet-visible dual-wavelength detection. To avoid cell aggregation or damage during electrophoresis, cells after staining were fixed with 4% formaldehyde and were continuously introduced into the capillary by EOF. The absorbance of a cell at 590 nm was used to determine its viability. An absorbance of two milli-absorbance unit at 590 nm was the clear cut-off point for living and dead Hela cells in our experiments. Good viability correlation between the conventional trypan blue staining assay and our established CE method (correlation coefficient, R(2)=0.9623) was demonstrated by analysis of cell mixtures with varying proportions of living and dead cells. The CE method was also used to analyze the cytotoxicity of methylmercury, and the results were in good agreement with the trypan blue staining assay and 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide methods. Compared with the 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide method, our established CE method can be easily automated to report cell viability based on the state of individual cells. Tedious manual cell counting and human error due to investigator bias can be avoided by using this method.
Ren
Xiaomin
X
School of Life Science, Beijing Institute of Technology, Beijing, PR China.
Qu
Feng
F
Zhang
Lu
L
Ding
Jinmei
J
Liu
Qingsheng
Q
D002470:Cell Survival / Q000502:physiology*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D015203:Reproducibility of Results; D014343:Trypan Blue / Q000378:metabolism
D014343:Trypan Blue