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Localizing sites of intradendritic electrophysiological recordings by confocal light microscopy
Author(s) -
Smith Karen L.,
Turner James N.,
Szarowski Donald H.,
Swann John W.
Publication year - 1994
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.1070290408
Subject(s) - lucifer yellow , microelectrode , confocal , electrophysiology , confocal microscopy , multielectrode array , neurophysiology , neuroscience , anatomy , chemistry , electrode , biology , optics , intracellular , physics , biochemistry , gap junction
Studies were undertaken to develop microscopic methods and imaging procedures that would permit identification of sites of intradendritic microelectrode recordings from pyramidal cells in hippocampal slice preparations. Intradendritic recording were obtained with sharp microelectrodes filled with the dye lucifer yellow. Following a recording session a neuron was iontophoretically injected with the dye and imaged by fluorescence videomicroscopy. Images were stored on videotape for later analysis. They provided a record of the location of the microelectrode recording site. After withdrawal of the microelectrode, slices were processed histologically and imaged a second time with a Bio‐Rad 600 confocal attachment on an Olympus BH‐2 microscope. Confocal images provided detailed anatomical information in three dimensions. In most instances, a clear identification of the recording site was achieved by comparing video images containing the recording electrode and confocal images. Neurophysiological recordings obtained from proximal and distal apical dendrites were markedly different. Proximal dendritic recordings were similar to those obtained from pyramidal cell soma. However, distal dendrites were not electroresponsive when depolarized by intracellular current injection. The techniques described here, or variations that employ patch electrodes, could provide valuable information that should further an understanding of the properties of dendrites in the central nervous system. © 1994 Wiley‐Liss, Inc.