Share Email Print

Proceedings Paper

A setup for combined multiphoton laser scanning microscopic and multi-electrode patch clamp experiments on brain slices
Author(s): P. Johannes Helm; Trond Reppen; Paul Heggelund
Format Member Price Non-Member Price
PDF $17.00 $21.00

Paper Abstract

Multi Photon Laser Scanning Microscopy (MPLSM) appears today as one of the most powerful experimental tools in cellular neurophysiology, notably in studies of the functional dynamics of signal processing in single neurons. Simultaneous recording of fluorescence signals at high spatial and temporal resolution and electric signals by means of multi electrode patch clamp techniques have provided new paths for the systematic investigation of neuronal mechanisms. In particular, this approach has opened for direct studies of dendritic signal processing in neurons. We report about a setup optimized for simultaneous electrophysiological multi electrode patch clamp and multi photon laser scanning fluorescence microscopic experiments on brain slices. The microscopic system is based on a modified commercially available confocal scanning laser microscope (CLSM). From a technical and operational point of view, two developments are important: Firstly, in order to reduce the workload for the experimentalist, who in general is forced to concentrate on controlling the electrophysiological parameters during the recordings, a system of shutters has been installed together with dedicated electronic modules protecting the photo detectors against destructive light levels caused by erroneous opening or closing of microscopic light paths by the experimentalist. Secondly, the standard detection unit has been improved by installing the photomultiplier tubes (PMT) in a Peltier cooled thermal box shielding the detector from both room temperature and distortions caused by external electromagnetic fields. The electrophysiological system is based on an industrial standard multi patch clamp unit ergonomically arranged around the microscope stage. The electrophysiological and scanning processes can be time coordinated by standard trigger electronics.

Paper Details

Date Published: 13 February 2009
PDF: 8 pages
Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 71832Q (13 February 2009); doi: 10.1117/12.810556
Show Author Affiliations
P. Johannes Helm, Univ. of Oslo (Norway)
Trond Reppen, Univ. of Oslo (Norway)
Paul Heggelund, Univ. of Oslo (Norway)

Published in SPIE Proceedings Vol. 7183:
Multiphoton Microscopy in the Biomedical Sciences IX
Ammasi Periasamy; Peter T. C. So, Editor(s)

© SPIE. Terms of Use
Back to Top
Sign in to read the full article
Create a free SPIE account to get access to
premium articles and original research
Forgot your username?