Instrumentation for Chemical Cytometry

Abstract

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.

Single-Cell Microbiology: Tools, Technologies, and Applications

“…Technical difficulties with whole-cell microcapillary separations may include the amphoteric nature of some cell types and problems with cell clumping, adhesion to capillary walls, or cell lysis (15,134).…”

“…Microcapillary-based methods for performing biochemical separations of the contents of individual cells, including yeast, on May 11, 2018 by guest http://mmbr.asm.org/ have also been described (98,132,133,134,139). Individual cells may be delivered into a capillary tube by suction forces or electroosmotic flow, with monitoring via microscopy (134,139).…”

“…Individual cells may be delivered into a capillary tube by suction forces or electroosmotic flow, with monitoring via microscopy (134,139). After a cell is loaded into the capillary, it is lysed to release the cell contents for analysis.…”

“…After a cell is loaded into the capillary, it is lysed to release the cell contents for analysis. The addition of surfactant may be sufficient to lyse mammalian cells, but thick-walled microorganisms such as S. cerevisiae may require spheroplasting first (134,139). Krylov et al (133) found that if electrophoresis was begun immediately following cell lysis, enzymatic activity was effectively "quenched" and artifacts stemming from nonmetabolic enzymatic activity or other degradative processes were suppressed.…”

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

“…At present, there is interest in single cell or subcellular analysis and therefore in developing the appropriate instrumentation and methods [170][171][172]. CE miniaturization will continue either through reducing column dimensions or placing entire electrophoresis systems on planar microfluidic chips [165,[173][174][175][176][177][178][179].…”