[Updates] 

July 2006. Roche Applied Science has stopped selling POD conjugated sheep anti-FITC fab fragments. DAKO distributes POD conjugated rabbit anti-FITC fab fragments (Cat # P5100), however, we have not yet tested it.

September 2004. Peter Vize's group has extended the use of fluorescent-tyramide reactions to immunohistochemistry applications in Xenopus and details synthesis and usage of Cy3-tyramide. Zhou and Vize (2004) Dev Biol 271(2):322-38. Pubmed

July 2004. We've published an additional paper using fluorescent in situ as well as fluorescent-tyramide amplified immunohistochemistry: Davidson, Keller, and DeSimone (2004) Gene Expression Patterns 4(4):457-66. Pubmed

[Old Information]

These pages document the fluorescent in situ techniques used in the paper:

Davidson, L. A., and Keller, R. E. (1999). Neural tube closure in Xenopus laevis involves medial migration, directed protrusive activity, cell intercalation, and convergent extension. Development, 126:4547-4556. Pubmed

The development of the technique sprang from my own ineptness with conventional wax-based histology and a pressing need to "see" 3-dimensional images of gene expression.

From first hand experience, I recommend the NEN Life Science's Red-FISH and Green-FISH kits (available as of 2006 from Invitrogen), epifluorescence capable stereomicroscopes from Zeiss, Leica, and Olympus, and "personal" confocal scanning microscopes from Olympus, Nikon, and Leica. The availablity of these reagents and instruments make fluorescent in situs possible and useful.

If you happen to make improvements to the protocols, or want to share results from your own troubleshooting please pass your ideas along and I'll include them in these pages.

Good Luck,

Lance Davidson

-PS I will be happy to lend whatever assistance I can, however, I am a mere junior faculty, so, before emailing me please fully investigate these pages and tips.