| Diolistic labeling of neuronal cultures and intact tissue using a hand-held gene gun. | |
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MedLine Citation:
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PMID: 17406443 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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John A O'Brien; Sarah C R Lummis |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Nature protocols Volume: 1 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2006 |
Date Detail:
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Created Date: 2007-04-04 Completed Date: 2007-06-28 Revised Date: 2013-03-27 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1517-21 Citation Subset: IM |
Affiliation:
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Neurobiology Division, MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Biolistics
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methods* Carbocyanines Cell Culture Techniques / methods* Cell Line Humans Neurons / cytology* Staining and Labeling / methods* |
| Grant Support | |
ID/Acronym/Agency:
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081925//Wellcome Trust; //Wellcome Trust |
| Chemical | |
Reg. No./Substance:
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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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