Multicolor Time-lapse Imaging of Transgenic Zebrafish: Visualizing Retinal Stem Cells Activated by Targeted Neuronal Cell Ablation

Hdl Handle:
http://hdl.handle.net/10675.2/669
Title:
Multicolor Time-lapse Imaging of Transgenic Zebrafish: Visualizing Retinal Stem Cells Activated by Targeted Neuronal Cell Ablation
Authors:
Ariga, Junko; Walker, Steven L.; Mumm, Jeff S.
Abstract:
High-resolution time-lapse imaging of living zebrafish larvae can be utilized to visualize how biological processes unfold (for review see 1). Compound transgenic fish which express different fluorescent reporters in neighboring cell types provide a means of following cellular interactions 2 and/or tissue-level responses to experimental manipulations over time. In this video, we demonstrate methods that can be used for imaging multiple transgenically labeled cell types serially in individual fish over time courses that can span from minutes to several days. The techniques described are applicable to any study seeking to correlate the "behavior" of neighboring cells types over time, including: 1) serial 'catch and release' methods for imaging a large number of fish over successive days, 2) simplified approaches for separating fluorophores with overlapping excitation/emission profiles (e.g., GFP and YFP), 3) use of hypopigmented mutant lines to extend the time window available for high-resolution imaging into late larval stages of development, 4) use of membrane targeted fluorescent reporters to reveal fine morphological detail of individual cells as well as cellular details in larger populations of cells, and 5) a previously described method for chemically-induced ablation of transgenically targeted cell types; i.e., nitroreductase (NTR) mediated conversion of prodrug substrates, such as metronidazole (MTZ), to cytotoxic derivatives 3,5.
Citation:
J Vis Exp. 2010 Sep 20;(43):2093
Issue Date:
20-Sep-2010
URI:
http://hdl.handle.net/10675.2/669
DOI:
10.3791/2093
PubMed ID:
20972390
PubMed Central ID:
PMC3157880
Type:
Article
ISSN:
1940-087X
Appears in Collections:
Department of Cellular Biology and Anatomy: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorAriga, Junkoen_US
dc.contributor.authorWalker, Steven L.en_US
dc.contributor.authorMumm, Jeff S.en_US
dc.date.accessioned2012-10-26T16:29:28Z-
dc.date.available2012-10-26T16:29:28Z-
dc.date.issued2010-09-20en_US
dc.identifier.citationJ Vis Exp. 2010 Sep 20;(43):2093en_US
dc.identifier.issn1940-087Xen_US
dc.identifier.pmid20972390en_US
dc.identifier.doi10.3791/2093en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/669-
dc.description.abstractHigh-resolution time-lapse imaging of living zebrafish larvae can be utilized to visualize how biological processes unfold (for review see 1). Compound transgenic fish which express different fluorescent reporters in neighboring cell types provide a means of following cellular interactions 2 and/or tissue-level responses to experimental manipulations over time. In this video, we demonstrate methods that can be used for imaging multiple transgenically labeled cell types serially in individual fish over time courses that can span from minutes to several days. The techniques described are applicable to any study seeking to correlate the "behavior" of neighboring cells types over time, including: 1) serial 'catch and release' methods for imaging a large number of fish over successive days, 2) simplified approaches for separating fluorophores with overlapping excitation/emission profiles (e.g., GFP and YFP), 3) use of hypopigmented mutant lines to extend the time window available for high-resolution imaging into late larval stages of development, 4) use of membrane targeted fluorescent reporters to reveal fine morphological detail of individual cells as well as cellular details in larger populations of cells, and 5) a previously described method for chemically-induced ablation of transgenically targeted cell types; i.e., nitroreductase (NTR) mediated conversion of prodrug substrates, such as metronidazole (MTZ), to cytotoxic derivatives 3,5.en_US
dc.rightsCopyright © 2010, Journal of Visualized Experimentsen_US
dc.subjectNeuroscienceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAnimals, Genetically Modifieden_US
dc.subject.meshCell Lineen_US
dc.subject.meshFluorescent Dyesen_US
dc.subject.meshImage Processing, Computer-Assisteden_US
dc.subject.meshNerve Regenerationen_US
dc.subject.meshNeuronsen_US
dc.subject.meshRetinaen_US
dc.subject.meshStem Cellsen_US
dc.subject.meshZebrafishen_US
dc.titleMulticolor Time-lapse Imaging of Transgenic Zebrafish: Visualizing Retinal Stem Cells Activated by Targeted Neuronal Cell Ablationen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC3157880en_US
dc.contributor.corporatenameDepartment of Cellular Biology and Anatomy-

Related articles on PubMed

All Items in Scholarly Commons are protected by copyright, with all rights reserved, unless otherwise indicated.