Fluorescent In-Situs and FCIS (Vize lab): Difference between revisions

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[[File:33ffar.jpg|200px|thumb|left|FISH, Fluorescein]]
[[File:2876.jpg|200px|thumb|left|FCIS; fluorescent and colorimetric in situ]]
[[File:38-doubleFISH-big.jpg|200px|thumb|right|Double FISH, Fluorescein and Cy3]]
[[File:frog10x-big.jpg|200px|thumb|right|FISH plus transmitted light]]
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.
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.


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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
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:'''
'''Papers and other sites:'''



Revision as of 13:36, 15 November 2011

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.

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:

Papers and other sites:

Zhou, X. and Vize, P.D. (2004). Proximo-distal specialization of epithelial transport processes within the Xenopus pronephric tubules. Developmental Biology 271: 322-338.

Lance Davidson's flourescent in situ methods page