Automated capillary electrophoresis system for fast single-cell analysis.

Area of Science:

• Analytical Chemistry
• Biochemistry
• Cell Biology

Background:

• Capillary electrophoresis (CE) offers potential for single-cell analysis but is hindered by low throughput.
• Automated platforms are needed to overcome limitations in analyzing biological samples at the single-cell level.

Purpose of the Study:

• To develop and validate an automated platform for high-throughput single-cell capillary electrophoresis.
• To assess the platform's ability to measure cellular enzyme activity and identify subpopulations.

Main Methods:

• An automated platform integrating microfabricated cell traps and a three-channel system for rapid buffer exchange was developed.
• Single-cell capillary electrophoresis was performed on cells labeled with fluorescent markers and substrates.
• Protein kinase B (PKB) and sphingosine kinase (SK) activities were measured in single cells.

Main Results:

• The platform achieved a throughput of 3.5 cells/min for general analysis and 2.1 cells/min for sphingosine kinase activity measurements.
• PKB activity remained unchanged, indicating no perturbation of basal cell physiology.
• Eighty-eight percent of leukemic cells exhibited upregulated SK activity, with distinct subpopulations identified.

Conclusions:

• The developed automated CE platform significantly increases throughput for single-cell analysis.
• This system enables the study of cellular enzyme activity and the identification of cell subpopulations with distinct biological phenomena.
• The platform demonstrates stable and reproducible separations, facilitating novel biological discoveries.