Document Detail


Diolistic labeling of neuronal cultures and intact tissue using a hand-held gene gun.
MedLine Citation:
PMID:  17406443     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Diolistic labeling is a highly efficient method for introducing dyes into cells using biolistic techniques. The use of lipophilic carbocyanine dyes, combined with particle-mediated biolistic delivery using a hand-held gene gun, allows non-toxic labeling of multiple cells in both living and fixed tissue. The technique is rapid (labeled cells can be visualized in minutes) and technically undemanding. Here, we provide a detailed protocol for diolistic labeling of cultured human embryonic kidney 293 cells and whole brain using a hand-held gene gun. There are four major steps: (i) coating gold microcarriers with one or more dyes; (ii) transferring the microcarriers into a cartridge to make a bullet; (iii) preparation of cells or intact tissue; and (iv) firing the microcarriers into cells or tissue. The method can be readily adapted to other cell types and tissues. This protocol can be completed in less than 1 h.
Authors:
John A O'Brien; Sarah C R Lummis
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  1     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2006  
Date Detail:
Created Date:  2007-04-04     Completed Date:  2007-06-28     Revised Date:  2014-02-19    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1517-21     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Biolistics / methods*
Carbocyanines
Cell Culture Techniques / methods*
Cell Line
Humans
Neurons / cytology*
Staining and Labeling / methods*
Grant Support
ID/Acronym/Agency:
081925//Wellcome Trust; MC_U105184294//Medical Research Council; //Wellcome Trust
Chemical
Reg. No./Substance:
0/Carbocyanines; 84109-08-0/carbocyanine dye DiIC12(3)
Comments/Corrections

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


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