I stained zebrafish larvae with double staining solution but the alcian blue stains more and I can see only the bluish larvae. How can I destain them?
We have been doing bone n cartilage staining using alcian blue and alzarin red...wat protocol r u using...generally we follwo a 7day protocol n d destaining steps include overnight incubation in pbs n alcohol solution...let me know ur process i will give u the precise volume and d composition
I am following the procedure as described in walker & kimmel 2007. fixing larvae in 4% PFA for 2 hours and dehydrating for 10 min in 50% alcohol. Then we add staining solution (1:100 ratio of Sol A and Sol B) and keep it for over night. ( Sol A - 0.02% alcian blue, 125mM MgCl, 70% ethanol), (Sol B - 0.5% alizarin red). After staining we are bleaching with hydrogen peroxide for 20 min with KOH. Then we are clearing with increasing gradient of KOH and glycerol for overnight. Totally it took 3 days to finish this procedure. But when staining, I think, I add more alcian blue. I need to destain to recover the sample. Can you help me?
See the volume may be a hinderance over here as u said...we do in 96 well plate..completing immersing the embryo in it....and then follow the protocol..i will give u d detailed thing soon...
I use eppendorfs and i can get good staining. But this time, I did a mistake of adding more alcian blue. How to destain the alcian blue dye?
and, thank you for your suggestions.
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