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Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology

Monitoring movements of larval Drosophila with electrical detection allows one to record the behaviors without the use of lights and cameras. This is a suitable technique when studying the use of light-sensitive proteins in optogenetic studies. Electrical measures are feasible to use in determining...

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Autores principales: de Castro, Noah, Cooper, Robin Lewis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7189670/
https://www.ncbi.nlm.nih.gov/pubmed/31991683
http://dx.doi.org/10.3390/mps3010012
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author de Castro, Noah
Cooper, Robin Lewis
author_facet de Castro, Noah
Cooper, Robin Lewis
author_sort de Castro, Noah
collection PubMed
description Monitoring movements of larval Drosophila with electrical detection allows one to record the behaviors without the use of lights and cameras. This is a suitable technique when studying the use of light-sensitive proteins in optogenetic studies. Electrical measures are feasible to use in determining when a larva starts to move or continues to move after a light induced activation of channelrhodopsin. We have developed a technique using an electrical measure of the media as an index of larval movement. As a proof of concept, recordings with an infrared camera of the larval movement were simultaneous made with electrical measures. The two techniques parallel each other in their ability to index larval movements. Bright light-emitting diode (LED) lights used in optogenetic experiments tend to saturate the detectors of the camera unless filters are used and different filters maybe necessary depending on the LED spectrum and sensitivity of the camera. Impedance measures are independent of the type of LED or brightness. We also assessed the use of a non-solvent based glue (3M Vetbond) to hold larvae in place while measuring synaptic function of neuromuscular junctions, cardiac function and influence of modulators, or activation of light-sensitive channels.
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spelling pubmed-71896702020-05-01 Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology de Castro, Noah Cooper, Robin Lewis Methods Protoc Protocol Monitoring movements of larval Drosophila with electrical detection allows one to record the behaviors without the use of lights and cameras. This is a suitable technique when studying the use of light-sensitive proteins in optogenetic studies. Electrical measures are feasible to use in determining when a larva starts to move or continues to move after a light induced activation of channelrhodopsin. We have developed a technique using an electrical measure of the media as an index of larval movement. As a proof of concept, recordings with an infrared camera of the larval movement were simultaneous made with electrical measures. The two techniques parallel each other in their ability to index larval movements. Bright light-emitting diode (LED) lights used in optogenetic experiments tend to saturate the detectors of the camera unless filters are used and different filters maybe necessary depending on the LED spectrum and sensitivity of the camera. Impedance measures are independent of the type of LED or brightness. We also assessed the use of a non-solvent based glue (3M Vetbond) to hold larvae in place while measuring synaptic function of neuromuscular junctions, cardiac function and influence of modulators, or activation of light-sensitive channels. MDPI 2020-01-24 /pmc/articles/PMC7189670/ /pubmed/31991683 http://dx.doi.org/10.3390/mps3010012 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
de Castro, Noah
Cooper, Robin Lewis
Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title_full Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title_fullStr Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title_full_unstemmed Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title_short Impedance Measures and a Mounting Technique for Drosophila: Larval Movements, Heart Rate, Imaging, and Electrophysiology
title_sort impedance measures and a mounting technique for drosophila: larval movements, heart rate, imaging, and electrophysiology
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7189670/
https://www.ncbi.nlm.nih.gov/pubmed/31991683
http://dx.doi.org/10.3390/mps3010012
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