Patch-clamp recording in brain slices with improved slicer technology

Geiger, J. R. P., Bischofberger, J., Vida, I., Fröbe, U., Pfitzinger, S., Weber, H. J.,, Haverkampf, K. and Jonas, P. (2002) Patch-clamp recording in brain slices with improved slicer technology. Pflügers Archiv - European Journal of Physiology, 443(3), pp. 491-501. (doi: 10.1007/s00424-001-0735-3)

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Abstract

The use of advanced patch-clamp recording techniques in brain slices, such as simultaneous recording from multiple neurons and recording from dendrites or presynaptic terminals, demands slices of the highest quality. In this context the mechanics of the tissue slicer are an important factor. Ideally, a tissue slicer should generate large-amplitude and high-frequency movements of the cutting blade in a horizontal axis, with minimal vibrations in the vertical axis. We developed a vibroslicer that fulfils these in part conflicting requirements. The oscillator is a permanent-magnet-coil-leaf-spring system. Using an auto-resonant mechano-electrical feedback circuit, large horizontal oscillations (up to 3 mm peak-to-peak) with high frequency (≈90 Hz) are generated. To minimize vertical vibrations, an adjustment mechanism was employed that allowed alignment of the cutting edge of the blade with the major axis of the oscillation. A vibroprobe device was used to monitor vertical vibrations during adjustment. The system is based on the shading of the light path between a light-emitting diode (LED) and a photodiode. Vibroprobe monitoring revealed that the vibroslicer, after appropriate adjustment, generated vertical vibrations of <1 µm, significantly less than many commercial tissue slicers. Light- and electron-microscopic analysis of surface layers of slices cut with the vibroslicer showed that cellular elements, dendritic processes and presynaptic terminals are well preserved under these conditions, as required for patch-clamp recording from these structures.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Vida, Dr Imre
Authors: Geiger, J. R. P., Bischofberger, J., Vida, I., Fröbe, U., Pfitzinger, S., Weber, H. J.,, Haverkampf, K., and Jonas, P.
College/School:College of Medical Veterinary and Life Sciences > School of Life Sciences
Journal Name:Pflügers Archiv - European Journal of Physiology
Publisher:Springer-Verlag
ISSN:0031-6768
ISSN (Online):1432-2013
Published Online:05 March 2014

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