Cargando…

WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans

BACKGROUND: There are four main phenotypes that are assessed in whole organism studies of Caenorhabditis elegans; mortality, movement, fecundity and size. Procedures have been developed that focus on the digital analysis of some, but not all of these phenotypes and may be limited by expense and limi...

Descripción completa

Detalles Bibliográficos
Autores principales: Mathew, Mark D., Mathew, Neal D., Ebert, Paul R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3311640/
https://www.ncbi.nlm.nih.gov/pubmed/22457766
http://dx.doi.org/10.1371/journal.pone.0033483
_version_ 1782227801069322240
author Mathew, Mark D.
Mathew, Neal D.
Ebert, Paul R.
author_facet Mathew, Mark D.
Mathew, Neal D.
Ebert, Paul R.
author_sort Mathew, Mark D.
collection PubMed
description BACKGROUND: There are four main phenotypes that are assessed in whole organism studies of Caenorhabditis elegans; mortality, movement, fecundity and size. Procedures have been developed that focus on the digital analysis of some, but not all of these phenotypes and may be limited by expense and limited throughput. We have developed WormScan, an automated image acquisition system that allows quantitative analysis of each of these four phenotypes on standard NGM plates seeded with E. coli. This system is very easy to implement and has the capacity to be used in high-throughput analysis. METHODOLOGY/PRINCIPAL FINDINGS: Our system employs a readily available consumer grade flatbed scanner. The method uses light stimulus from the scanner rather than physical stimulus to induce movement. With two sequential scans it is possible to quantify the induced phototactic response. To demonstrate the utility of the method, we measured the phenotypic response of C. elegans to phosphine gas exposure. We found that stimulation of movement by the light of the scanner was equivalent to physical stimulation for the determination of mortality. WormScan also provided a quantitative assessment of health for the survivors. Habituation from light stimulation of continuous scans was similar to habituation caused by physical stimulus. CONCLUSIONS/SIGNIFICANCE: There are existing systems for the automated phenotypic data collection of C. elegans. The specific advantages of our method over existing systems are high-throughput assessment of a greater range of phenotypic endpoints including determination of mortality and quantification of the mobility of survivors. Our system is also inexpensive and very easy to implement. Even though we have focused on demonstrating the usefulness of WormScan in toxicology, it can be used in a wide range of additional C. elegans studies including lifespan determination, development, pathology and behavior. Moreover, we have even adapted the method to study other species of similar dimensions.
format Online
Article
Text
id pubmed-3311640
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-33116402012-03-28 WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans Mathew, Mark D. Mathew, Neal D. Ebert, Paul R. PLoS One Research Article BACKGROUND: There are four main phenotypes that are assessed in whole organism studies of Caenorhabditis elegans; mortality, movement, fecundity and size. Procedures have been developed that focus on the digital analysis of some, but not all of these phenotypes and may be limited by expense and limited throughput. We have developed WormScan, an automated image acquisition system that allows quantitative analysis of each of these four phenotypes on standard NGM plates seeded with E. coli. This system is very easy to implement and has the capacity to be used in high-throughput analysis. METHODOLOGY/PRINCIPAL FINDINGS: Our system employs a readily available consumer grade flatbed scanner. The method uses light stimulus from the scanner rather than physical stimulus to induce movement. With two sequential scans it is possible to quantify the induced phototactic response. To demonstrate the utility of the method, we measured the phenotypic response of C. elegans to phosphine gas exposure. We found that stimulation of movement by the light of the scanner was equivalent to physical stimulation for the determination of mortality. WormScan also provided a quantitative assessment of health for the survivors. Habituation from light stimulation of continuous scans was similar to habituation caused by physical stimulus. CONCLUSIONS/SIGNIFICANCE: There are existing systems for the automated phenotypic data collection of C. elegans. The specific advantages of our method over existing systems are high-throughput assessment of a greater range of phenotypic endpoints including determination of mortality and quantification of the mobility of survivors. Our system is also inexpensive and very easy to implement. Even though we have focused on demonstrating the usefulness of WormScan in toxicology, it can be used in a wide range of additional C. elegans studies including lifespan determination, development, pathology and behavior. Moreover, we have even adapted the method to study other species of similar dimensions. Public Library of Science 2012-03-23 /pmc/articles/PMC3311640/ /pubmed/22457766 http://dx.doi.org/10.1371/journal.pone.0033483 Text en Mathew et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Mathew, Mark D.
Mathew, Neal D.
Ebert, Paul R.
WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title_full WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title_fullStr WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title_full_unstemmed WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title_short WormScan: A Technique for High-Throughput Phenotypic Analysis of Caenorhabditis elegans
title_sort wormscan: a technique for high-throughput phenotypic analysis of caenorhabditis elegans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3311640/
https://www.ncbi.nlm.nih.gov/pubmed/22457766
http://dx.doi.org/10.1371/journal.pone.0033483
work_keys_str_mv AT mathewmarkd wormscanatechniqueforhighthroughputphenotypicanalysisofcaenorhabditiselegans
AT mathewneald wormscanatechniqueforhighthroughputphenotypicanalysisofcaenorhabditiselegans
AT ebertpaulr wormscanatechniqueforhighthroughputphenotypicanalysisofcaenorhabditiselegans