Kaede on fixed specimens

Posted by xavier Sanjuan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Overnight-scan-of-human-fibroblasts-tp2543999p2545160.html

Hi list,

 

One of my users is using Kaede in a zebrafish model to run in vivo experiments. Once they have finished he fixes the embryo to do further imaging (also to attempt FISH experiments), but whereas the green signal can be imaged without any problem, the red fluorescence in the photoconverted areas vanishes 1-2 days after mounting. He is doing a standard 4%PFA fixation, and he mounts the zebrafish embryos with glycerol.

 

Is he doing something wrong that is bleaching/destroying the photoconverted Kaede?

 

Thanks in advance for your help!

 

Best,

 

Xavi.

 

___________________________________

Xavier Sanjuan
Servei de Microscòpia Confocal
Departament de Ciències Experimentals i de la Salut
Universitat Pompeu Fabra
Parc de Recerca Biomèdica de Barcelona
Doctor Aiguader, 88 - Sala 309
08003 Barcelona - Spain

Tel.:  + 34 93 316 08 64
Fax: + 34 93 316 09 01
E-mail: [hidden email]
Web: http://www.upf.edu/sct