TY - JOUR
T1 - Simultaneous optical recording of evoked and spontaneous transients of membrane potential and intracellular calcium concentration with high spatio-temporal resolution
AU - Sinha, Saurabh R.
AU - Patel, Saumil S.
AU - Saggau, Peter
N1 - Funding Information:
We thank Drs. R. Gray, D. Johnston and W. Schilling for kindly allowing us to use some of their equipment, and A. Bullen and Drs. B. Christie and L.-G. Wu for critical and helpful comments on the manuscript. This work was supported by an NRSA predoctoral fellowship from NIMH to S.R.S. and by departmental funds.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1995/8
Y1 - 1995/8
N2 - We have developed a system for simultaneous optical recording of transients of membrane potential and intracellular calcium concentration from mammalian brain slice preparations with high spatio-temporal resolution. Simultaneous recording was achieved by using two dedicated photodetectors together with two fluorescent indicators. Specifically, the calcium-sensitive dye Calcium Orange and the voltage-sensitive dye RH-414 were selected because they have overlapping excitation spectra, but separable emission spectra. Transverse guinea pig hippocampal slices were double-loaded by bath application of the membrane-permeant form of Calcium Orange and RH-414. Transients of intracellular calcium concentration and membrane potential associated with evoked neural activity in hippocampal areas CA1 and CA3 were recorded. Furthermore, we have recorded calcium and voltage transients associated with spontaneous epileptiform activity induced by bath application of an epileptogenic drug, 4-aminopyridine. The use of photodiode matrices (10 × 10 elements each) as detectors gives the high spatial (200 × 200 μm/element with a 10 × objective) and temporal resolution (570 μs/frame). The recording system also includes a CCD camera for obtaining images of the preparation and overlaying the image with the optically detected signals. A software package has been developed for setting up the experimental protocol(s) and for collecting, processing, displaying, and analyzing the data in an user-friendly, windows-based environment.
AB - We have developed a system for simultaneous optical recording of transients of membrane potential and intracellular calcium concentration from mammalian brain slice preparations with high spatio-temporal resolution. Simultaneous recording was achieved by using two dedicated photodetectors together with two fluorescent indicators. Specifically, the calcium-sensitive dye Calcium Orange and the voltage-sensitive dye RH-414 were selected because they have overlapping excitation spectra, but separable emission spectra. Transverse guinea pig hippocampal slices were double-loaded by bath application of the membrane-permeant form of Calcium Orange and RH-414. Transients of intracellular calcium concentration and membrane potential associated with evoked neural activity in hippocampal areas CA1 and CA3 were recorded. Furthermore, we have recorded calcium and voltage transients associated with spontaneous epileptiform activity induced by bath application of an epileptogenic drug, 4-aminopyridine. The use of photodiode matrices (10 × 10 elements each) as detectors gives the high spatial (200 × 200 μm/element with a 10 × objective) and temporal resolution (570 μs/frame). The recording system also includes a CCD camera for obtaining images of the preparation and overlaying the image with the optically detected signals. A software package has been developed for setting up the experimental protocol(s) and for collecting, processing, displaying, and analyzing the data in an user-friendly, windows-based environment.
KW - Brain slice
KW - Calcium Orange
KW - Calcium indicator
KW - Epileptiform activity
KW - Fluorescence imaging
KW - Intracellular calcium
KW - Membrane potential
KW - RH-414
KW - Simultaneous recording
KW - Voltage-sensitive dye
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U2 - 10.1016/0165-0270(94)00219-7
DO - 10.1016/0165-0270(94)00219-7
M3 - Article
C2 - 8544487
AN - SCOPUS:0029127573
SN - 0165-0270
VL - 60
SP - 49
EP - 60
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
IS - 1-2
ER -