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Measuring ca(2+) oscillations in Mammalian eggs.
MedLine Citation:
PMID:  23138956     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
At fertilization mammalian eggs are activated by a prolonged series of oscillations in the intracellular free Ca(2+) concentration. These oscillations can be monitored with any number of Ca(2+)-sensitive fluorescent dyes. The oscillations last for several hours at fertilization and so there are some considerations with mammalian eggs that make them distinct from somatic cells that are commonly used in Ca(2+) imaging experiments. I describe the use of two particular dyes that can be loaded into mouse eggs and that give the most valuable results. The first one is PE3 which can be loaded by incubation with the AM form of the dye which is membrane permeable. The other is rhod dextran which requires microinjection. Either one of these dyes offers advantages over the more commonly used fura2. I describe the way that the fluorescence from dye-loaded eggs is measured with a conventional epifluorescence microscope and a CCD camera.
Authors:
Karl Swann
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  957     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-09     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  231-48     Citation Subset:  IM    
Affiliation:
School of Medicine, Cardiff University, Heath Park, Cardiff, UK, swannk1@cf.ac.uk.
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