This article demonstrates the dissection and incubation of rabbit retina and particle-mediated gene transfer of plasmids encoding GFP or a variety of subcellular markers into retinal ganglion cells.
Making gene gun bullets (GOLD)
Preparation of gene-gun
Preparation of media
1 bottle Ames solution (Sigma, 8.8g) |
1.9 gm sodium bicarbonate |
10 ml 1% pen/strep/L-Glutamine (1:100) (Gibco/Invitrogen) |
990 ml distilled water to 1L |
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1 Liter |
485 ml of filtered dissection media |
5 ml of 1% N2 supplement (Gibco/Invitrogen) |
10ml 1% horse serum (Sigma) |
500 µl of phenol red |
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0.5 L |
Preparation of incubation chambers for retina
Preparation of rabbit retina
Gene gunning and incubation of retina
1. What can I do with this method?
2. How long can I keep adult retina?
3. How do I make sure your retina is healthy after this time?
4. Why do you want to manipulate only a few cells in the retina at a time?
5. Is this method suitable as a population stain?
6. Are there any “tricks” I have to know to make it work?
7. Could you gene-gun other stuff, rather than plasmids?
8. What are the limitations?
9. What other researcher’s work should one be aware of?