Document Detail


Fast Fos: rapid protocols for single- and double-labeling c-Fos immunohistochemistry in fresh frozen brain sections.
MedLine Citation:
PMID:  15585284     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Immunohistochemical localization of c-Fos immunoreactivity has been used successfully for over a decade to visualize patterns of neuronal activity in the brain and spinal cord. These experiments are extremely useful in identifying physiological or pharmacological activation of specific populations of neurons. Unfortunately, conventional c-Fos immunohistochemical protocols are very time and resource intensive. We have adapted and optimized established c-Fos immunohistochemistry (IHC) methodologies for use with fresh frozen brain tissue mounted directly onto slides. The resultant rapid protocols, which we refer to as "Fast Fos", include applications for single- and double-labeling, utilizing either enzyme-substrate or fluorescent detection systems. These protocols provide increased assay throughput and reproducibility, which can be further enhanced by use of an automated slide stainer. Taken as a whole, the c-Fos IHC protocols described in this report provide a flexible system for the identification of neuronal activation that substantially reduces time and resource expenditure while increasing quality and reproducibility of data.
Authors:
Sarah J Sundquist; Laura K Nisenbaum
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  141     ISSN:  0165-0270     ISO Abbreviation:  J. Neurosci. Methods     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2004-12-08     Completed Date:  2005-04-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  9-20     Citation Subset:  IM    
Affiliation:
Neuroscience Discovery Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285-0438, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Brain / cytology,  metabolism*
Fluorescent Antibody Technique / methods
Frozen Sections / methods
Immunohistochemistry / methods*
Intracellular Signaling Peptides and Proteins / analysis
Male
Neuropeptides / analysis
Proto-Oncogene Proteins c-fos / analysis*
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Software Design
Staining and Labeling / methods*
Time Factors
Tissue Fixation / methods
Chemical
Reg. No./Substance:
0/Intracellular Signaling Peptides and Proteins; 0/Neuropeptides; 0/Proto-Oncogene Proteins c-fos; 0/orexins

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