Document Detail


Measurement of neuronal Ca2+ transients using simultaneous microfluorimetry and electrophysiology.
MedLine Citation:
PMID:  3174384     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fluorescent indicator molecules, such as fura-2, are useful probes for studying the concentrations of ions in single cells. A key feature of these fluorescent dyes is the shift in their excitation spectra upon binding the ion, thus making alternate excitation from two wavelengths desirable. In this report we describe an inexpensive system for making simultaneous electrophysiological and dual excitation fluorescence measurements using equipment much of which is available in a typical biophysical laboratory. In order to synchronize the fluorescence signal with the appropriate excitation wavelength we devised a simple computer program which uses the output of photodiodes placed in the excitation beam to determine which wavelength is illuminating the cell. We also describe the use of a liquid light guide to transmit excitation illumination from the light source to the epifluorescence port of the microscope in order to isolate a perfusion chamber from light, electrical noise and vibration. A sensitive light detection system was assembled using a photomultiplier tube and discriminator that took advantage of sampling single unit events obtained with photon counting rather than the analog of anode current. However, rather than employing a sophisticated and expensive photon counting system, a filter was used to integrate the signal so that an analog output could be presented to a multichannel analog to digital converter commonly used for electrophysiological recordings.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
S A Thayer; M Sturek; R J Miller
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Pflügers Archiv : European journal of physiology     Volume:  412     ISSN:  0031-6768     ISO Abbreviation:  Pflugers Arch.     Publication Date:  1988 Jul 
Date Detail:
Created Date:  1988-11-07     Completed Date:  1988-11-07     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0154720     Medline TA:  Pflugers Arch     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  216-23     Citation Subset:  IM    
Affiliation:
Department of Pharmacological and Physiological Sciences, University of Chicago, IL 60637.
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MeSH Terms
Descriptor/Qualifier:
Animals
Benzofurans / diagnostic use
Calcium / analysis*,  physiology
Cytophotometry / instrumentation,  methods*
Electrophysiology / instrumentation,  methods*
Fura-2
Ganglia, Sympathetic / analysis,  physiology
Neurons, Afferent / analysis*,  physiology
Rats
Grant Support
ID/Acronym/Agency:
DA02121/DA/NIDA NIH HHS; DA02575/DA/NIDA NIH HHS; MH40165/MH/NIMH NIH HHS
Chemical
Reg. No./Substance:
0/Benzofurans; 7440-70-2/Calcium; 96314-98-6/Fura-2

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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