Instrumentation for chemical cytometry.
Capillary electrophoresis is ideally suited to chemical analysis of individual cells. Small mammalian somatic cells (approximately 15 microns in diameter) can be analyzed by injecting the intact cell into a capillary, lysing the cell, separating and detecting the cellular components, and reconditioning the capillary prior to the next injection. In this paper, we report on technical improvements to single-cell analysis. We designed an inexpensive multipurpose single-cell injector that facilitates the following: (i) monitoring of injection, (ii) reproducible pressure- or electrokinetic-driven injection of the cell, (iii) complete cell lysis by SDS within 30 s of injection, and (iv) pressure-driven capillary reconditioning. Furthermore, we report on the analysis of glycosylation and glycolysis in single human carcinoma cells (HT29 cell line). The reliability and quality of the analysis is confirmed by comparing electropherograms from single cells and those from purified cell extracts.
Krylov
S N
SN
Department of Chemistry, University of Alberta, Edmonton, Canada.
Starke
D A
DA
Arriaga
E A
EA
Zhang
Z
Z
Chan
N W
NW
Palcic
M M
MM
Dovichi
N J
NJ
D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006019:Glycolysis; D006031:Glycosylation; D019073:HT29 Cells; D006801:Humans; D008856:Microscopy, Fluorescence