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pH Change of buffer solution in a microcapillary chip and its suppression.
Akio Oki1, Yuzuru Takamura, Yoshitaka Ito
1Department of Materials Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan. oki@micro.mm.t.u-tokyo.ac.jp
Electrophoresis
|September 11, 2002
Summary
Electrolysis in microcapillary electrophoresis causes buffer solutions to become alkaline, inactivating cells. Two methods, salt bridges and electroosmotic flow pumps, effectively suppressed pH changes during B- and T-cell separation.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Cell Biology
Background:
- Microcapillary electrophoresis is used for separating B- and T-cells from lymphocytes.
- Electrolysis in small-volume reservoirs causes buffer solutions to become alkaline.
- This pH change inactivates cells during separation.
Purpose of the Study:
- To investigate the cause of cell inactivation during microcapillary electrophoresis.
- To develop methods for suppressing pH changes in microcapillary electrophoresis.
- To ensure stable conditions for B- and T-cell separation.
Main Methods:
- Utilized microcapillary chips with embedded ion-sensitive field-effect transistors (ISFET) to monitor pH.
- Implemented a salt bridge method to maintain buffer stability.
- Employed an electroosmotic flow (EOF) pump to neutralize alkaline solutions.
Main Results:
- Confirmed that electrolysis causes alkaline pH shifts in microcapillary electrophoresis reservoirs.
- Demonstrated that both salt bridges and EOF pumps effectively prevent pH changes.
- Achieved stable electrophoresis conditions for cell separation without pH fluctuation.
Conclusions:
- Electrolysis-induced pH changes are a critical issue in microcapillary electrophoresis of lymphocytes.
- Salt bridges and EOF pumps are viable solutions for maintaining stable buffer pH.
- These methods enable reliable separation and analysis of B- and T-cells.