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Whole-cell patch-clamp: true perforated or spontaneous conventional recordings?
1Department of Neurology, University of Rostock, Germany. ulf.strauss@med.uni-rostock.de
Pflugers Archiv : European Journal of Physiology
|August 21, 2001
Summary
Eosin dye effectively distinguishes perforated from conventional whole-cell patch-clamp recordings in lymphocytes. This method ensures accurate intracellular integrity preservation during electrophysiological studies.
Area of Science:
- Electrophysiology
- Cell Biology
- Biophysics
Background:
- Perforated whole-cell patch-clamp recording with nystatin is vital for maintaining intracellular integrity.
- Spontaneous shifts from perforated to conventional whole-cell configuration occur, particularly in lymphocytes.
- Existing electrophysiological criteria for perforated-patch configuration are insufficient.
Purpose of the Study:
- To evaluate the efficacy of eosin dye in differentiating between perforated and conventional whole-cell patch-clamp configurations.
- To establish a reliable method for verifying true perforated-patch recordings in lymphocytes.
Main Methods:
- Perforated whole-cell patch-clamp recordings were performed on rat T-lymphocytes.
- The pipette solution included the dye eosin.
- Electrophysiological recordings were analyzed in the presence of eosin.
Main Results:
- Eosin did not enter the cell during perforated whole-cell recordings.
- Cells in the conventional whole-cell configuration turned red immediately upon membrane rupture.
- Eosin exhibited minimal impact on the measured currents.
Conclusions:
- Eosin serves as a reliable indicator for confirming a genuine perforated-patch configuration.
- This dye facilitates accurate assessment of intracellular integrity during patch-clamp experiments.