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Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

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Glutaraldehyde enhanced dielectrophoretic yeast cell separation.

Zachary Gagnon¹, Jill Mazur, Hsueh-Chia Chang

¹Department of Chemical and Biomolecular Engineering, Center for Microfluidics and Medical Diagnostics, University of Notre Dame, Notre Dame, Indiana 46556, USA.

Biomicrofluidics

|March 11, 2010

Summary

Glutaraldehyde (GLT) selectively cross-links nonviable yeast cells, enhancing dielectrophoretic (DEP) separation. This method improves discrimination between viable and nonviable yeast by modifying DEP crossover frequency.

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Area of Science:

Biophysics
Cell Biology
Separation Science

Background:

Dielectrophoresis (DEP) is a label-free technique used for cell separation.
Viable and nonviable cells exhibit different electrical properties, but discrimination can be challenging.
Cell wall permeability differences between viable and nonviable yeast cells are not fully exploited for separation.

Purpose of the Study:

To develop an improved method for dielectrophoretic (DEP) discrimination and separation of viable and nonviable yeast cells.
To investigate the effect of glutaraldehyde (GLT) on cell electrical properties and DEP behavior.
To enhance the differences in DEP crossover frequency (cof) between viable and nonviable yeast cells.

Main Methods:

Measurement of DEP crossover frequency (cof) for viable and nonviable yeast cells across a range of buffer conductivities.
Selective cross-linking of nonviable yeast cells using glutaraldehyde (GLT).
Modeling of cell electrical properties using a dual-shelled oblate spheroid model fitted to cof data.

Main Results:

Glutaraldehyde (GLT) selectively cross-links nonviable yeast cells, significantly altering their DEP crossover frequency (cof).
Viable yeast cell cof remains largely unaffected by GLT treatment.
GLT treatment stabilizes cytoplasm conductivity in nonviable cells, minimizing ion leakage and increasing the difference in cof compared to viable cells.

Conclusions:

Glutaraldehyde (GLT) treatment effectively enhances dielectrophoretic (DEP) separation of viable and nonviable yeast cells.
Exploiting differences in cell wall permeability with GLT magnifies DEP effects for improved cell discrimination.
This method offers a promising approach for selective separation and analysis of cell populations.