|
imported>Xenbase gene generator |
| Line 1: |
Line 1: |
| These pages contain protocols for performing wholemount fluorescent in situs. Instructions for two-color FISH and for a combination of fluorescent and colorimetric in situs we have called FCIS. Data can be imaged with a fluorescence stereoscope or a confocal.
| | #REDIRECT [[XB-FEAT-980194]] |
| | |
| *[[FISH_/_Double_FISH|FISH protocol]]
| |
| *[[Flourescin_Tyramide_Synthesis|Fluorescein tyramide synthesis]]
| |
| *[[Cy3_Tyramide_Synthesis|Cy3 tyramide synthesis]]
| |
| *[[FCIS|FCIS protocol]]
| |
| | |
| FISH is not as sensitive as standard BM purple in situs, so if you have a very low abundance message it may not be detectable via fluorescence. You can however do a normal purple development for your low expression gene and a fluorescent counterstain- then overlay them to generate a FCIS image. These are both pretty and very good at highlighting quite subtle overlap
| |
| | |
| '''Images:'''
| |
| <gallery>
| |
| File:33ffar.jpg|FISH, Fluorescein
| |
| File:2876.jpg|FCIS; fluorescent and colorimetric in situ
| |
| File:38-doubleFISH-big.jpg|Double FISH, Fluorescein and Cy3
| |
| File:frog10x-big.jpg|FISH plus transmitted light
| |
| </gallery>
| |
| '''Papers and other sites:'''
| |
| | |
| Zhou, X. and Vize, P.D. (2004). Proximo-distal specialization of epithelial transport processes within the Xenopus pronephric tubules. [http://www.ncbi.nlm.nih.gov/pubmed/15223337?dopt=Abstract| Developmental Biology 271: 322-338.]
| |
| | |
| [http://www.faculty.virginia.edu/davidson/fluor_insitu/fluorescent_in_situ.html| Lance Davidson's flourescent in situ methods page]
| |
| | |
| Vize, P.D., McCoy, K.E., and Zhou, X. (2009). Multichannel wholemount fluorescent and fluorescent/chromogenic in situ hybridization in Xenopus embryos. [http://www.ncbi.nlm.nih.gov/pubmed/19498377| Nat Protoc. 4(6): 975-983.]
| |