Document Detail


Multi-position photoactivation and multi-time acquisition for large-scale cell tracing in avian embryos.
MedLine Citation:
PMID:  20516185     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Vertebrate development is a complex orchestration of cell and tissue movements. Tracing individual cell positions can rapidly become a large-scale problem because cell numbers often grow exponentially in the early embryo. A typical approach consists of fluorescently marking small numbers of cells within a large number of embryos, followed by comprehensive three-dimensional static or time-lapse imaging to map cell positions. However, for large-scale cell tracing, such as during organogenesis, the time, effort, and expense of this approach can be limiting. The multi-position acquisition method can be used to capture more than one location on a microscope stage and allow for multi-specimen imaging. When combined with photoactivation cell labeling, a tool for selective cell marking using laser excitation, multi-position imaging offers a powerful tool for rapid data acquisition. This protocol describes the technique and demonstrates its use to map cell movements in the chick spinal cord, using slice culture explants. The details of multiple slice culture preparation, multi-position photoactivation, and multi-time acquisition are described. These events are coordinated by setting up blocks of microscope instructions that execute sequentially. This method significantly decreases the time, effort, microscopy, and embryo costs by a factor of the number of specimens imaged per session, typically six.
Authors:
Joseph Steen; Jason A Morrison; Paul M Kulesa
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cold Spring Harbor protocols     Volume:  2010     ISSN:  1559-6095     ISO Abbreviation:  Cold Spring Harb Protoc     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-06-02     Completed Date:  2010-09-01     Revised Date:  2014-09-17    
Medline Journal Info:
Nlm Unique ID:  101524530     Medline TA:  Cold Spring Harb Protoc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  pdb.prot5447     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Calibration
Chick Embryo
Embryo, Nonmammalian / cytology*,  metabolism*,  radiation effects
Hot Temperature
Imaging, Three-Dimensional / methods*
Indicators and Reagents
Light*
Microinjections
Software
Staining and Labeling / methods*
Time Factors
Tissue Culture Techniques
Grant Support
ID/Acronym/Agency:
1R01HD057922/HD/NICHD NIH HHS; R01 HD057922/HD/NICHD NIH HHS; R01 HD057922-02/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
0/Indicators and Reagents
Comments/Corrections

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


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