Patch-clamp recording in brain slices with improved slicer technology Journal Article


Author(s): Geiger, Jörg R; Bischofberger, Joseph; Vida, Imre; Fröbe, Ulrich; Pfitzinger, S; Weber, H. J.; Haverkampf, Klaus; Jonas, Peter
Article Title: Patch-clamp recording in brain slices with improved slicer technology
Affiliation
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.
Keywords: Animals; Presynaptic Terminals/physiology; Microscopy, Electron; Patch-Clamp Techniques/methods; Dendrites/physiology; Mossy Fibers, Hippocampal/physiology; Microtomy/instrumentation; Mossy Fibers, Hippocampal/ultrastructure; Organ Culture Techniques/instrumentation
Journal Title: Pflugers Archiv : European Journal of Physiology
Volume: 443
Issue 3
ISSN: 0031-6768
Publisher: Springer  
Date Published: 2002-01-01
Start Page: 491
End Page: 501
DOI: 10.1007/s00424-001-0735-3
Open access: no