Voltage-Sensitive Dye Recording from Networks of Cultured Neurons

<p>This thesis describes the development and testing of a sensitive apparatus for recording electrical activity from microcultures of rat superior cervical ganglion (SCG) neurons by using voltage-sensitive fluorescent dyes.</p> <p>The apparatus comprises a feedback-regulated mer...

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Main Author: Chien, Chi-Bin
Format: Others
Language:en
Published: 1990
Online Access:https://thesis.library.caltech.edu/2517/1/Chien_cb_1990.pdf
Chien, Chi-Bin (1990) Voltage-Sensitive Dye Recording from Networks of Cultured Neurons. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/jwsd-2f67. https://resolver.caltech.edu/CaltechETD:etd-06082007-080125 <https://resolver.caltech.edu/CaltechETD:etd-06082007-080125>
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spelling ndltd-CALTECH-oai-thesis.library.caltech.edu-25172021-11-04T05:01:38Z https://thesis.library.caltech.edu/2517/ Voltage-Sensitive Dye Recording from Networks of Cultured Neurons Chien, Chi-Bin <p>This thesis describes the development and testing of a sensitive apparatus for recording electrical activity from microcultures of rat superior cervical ganglion (SCG) neurons by using voltage-sensitive fluorescent dyes.</p> <p>The apparatus comprises a feedback-regulated mercury arc light source, an inverted epifluorescence microscope, a novel fiber-optic camera with discrete photodiode detectors, and low-noise preamplifiers. Using an NA 0.75 objective and illuminating at 10 W/cm2 with the 546 nm mercury line, a typical SCG neuron stained with the styryl dye RH423 gives a detected photocurrent of 1 nA; the light source and optical detectors are quiet enough that the shot noise in this photocurrent--about.03% rms--dominates. The design, theory, and performance of this dye-recording apparatus are discussed in detail.</p> <p>Styryl dyes such as RH423 typically give signals of 1%/100 mV on these cells; the signals are linear in membrane potential, but do not appear to arise from a purely electrochromic mechanism. Given this voltage sensitivity and the noise level of the apparatus, it should be possible to detect both action potentials and subthreshold synaptic potentials from SCG cell bodies. In practice, dye recording can easily detect action potentials from every neuron in an SCG microculture, but small synaptic potentials are obscured by dye signals from the dense network of axons.</p> <p>In another microculture system that does not have such long and complex axons, this dye-recording apparatus should be able to detect synaptic potentials, making it possible to noninvasively map the synaptic connections in a microculture, and thus to study long-term synaptic plasticity.</p> 1990 Thesis NonPeerReviewed application/pdf en other https://thesis.library.caltech.edu/2517/1/Chien_cb_1990.pdf Chien, Chi-Bin (1990) Voltage-Sensitive Dye Recording from Networks of Cultured Neurons. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/jwsd-2f67. https://resolver.caltech.edu/CaltechETD:etd-06082007-080125 <https://resolver.caltech.edu/CaltechETD:etd-06082007-080125> https://resolver.caltech.edu/CaltechETD:etd-06082007-080125 CaltechETD:etd-06082007-080125 10.7907/jwsd-2f67
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language en
format Others
sources NDLTD
description <p>This thesis describes the development and testing of a sensitive apparatus for recording electrical activity from microcultures of rat superior cervical ganglion (SCG) neurons by using voltage-sensitive fluorescent dyes.</p> <p>The apparatus comprises a feedback-regulated mercury arc light source, an inverted epifluorescence microscope, a novel fiber-optic camera with discrete photodiode detectors, and low-noise preamplifiers. Using an NA 0.75 objective and illuminating at 10 W/cm2 with the 546 nm mercury line, a typical SCG neuron stained with the styryl dye RH423 gives a detected photocurrent of 1 nA; the light source and optical detectors are quiet enough that the shot noise in this photocurrent--about.03% rms--dominates. The design, theory, and performance of this dye-recording apparatus are discussed in detail.</p> <p>Styryl dyes such as RH423 typically give signals of 1%/100 mV on these cells; the signals are linear in membrane potential, but do not appear to arise from a purely electrochromic mechanism. Given this voltage sensitivity and the noise level of the apparatus, it should be possible to detect both action potentials and subthreshold synaptic potentials from SCG cell bodies. In practice, dye recording can easily detect action potentials from every neuron in an SCG microculture, but small synaptic potentials are obscured by dye signals from the dense network of axons.</p> <p>In another microculture system that does not have such long and complex axons, this dye-recording apparatus should be able to detect synaptic potentials, making it possible to noninvasively map the synaptic connections in a microculture, and thus to study long-term synaptic plasticity.</p>
author Chien, Chi-Bin
spellingShingle Chien, Chi-Bin
Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
author_facet Chien, Chi-Bin
author_sort Chien, Chi-Bin
title Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
title_short Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
title_full Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
title_fullStr Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
title_full_unstemmed Voltage-Sensitive Dye Recording from Networks of Cultured Neurons
title_sort voltage-sensitive dye recording from networks of cultured neurons
publishDate 1990
url https://thesis.library.caltech.edu/2517/1/Chien_cb_1990.pdf
Chien, Chi-Bin (1990) Voltage-Sensitive Dye Recording from Networks of Cultured Neurons. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/jwsd-2f67. https://resolver.caltech.edu/CaltechETD:etd-06082007-080125 <https://resolver.caltech.edu/CaltechETD:etd-06082007-080125>
work_keys_str_mv AT chienchibin voltagesensitivedyerecordingfromnetworksofculturedneurons
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