Diolistic labeling of neuronal cultures and intact tissue using a hand-held gene gun

• Published in: Nature protocols Vol. 1; no. 3; pp. 1517 - 1521
• Main Authors: Lummis, Sarah C R, O'Brien, John A
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01-11-2006
• Subjects: Biolistics - methods; Carbocyanines; Cell culture; Cell Culture Techniques - methods; Cell Line; Dyes; Dyes and dyeing; Humans; Methods; Neurons - cytology; Nucleic acid hybridization; Staining and Labeling - methods; Tissues; United Kingdom
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/nprot.2006.258

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.