Is there anyone who has tried to label cells with Hoechst or similar in living invertebrates? I am thinking about labeling the nuclei in the brain of a living insect. Has anyone ever tried it?
I stained zebrafish larvae with hoechst and DRAQ5 and it worked well. The fish survived it without any problems.
and you just dissect under anestesia (i would do it on bees) until you expose the tissue, drop some microliters of hoechst on it and after a few minutes all nuclei are stained like in fixed tissues? do you have an image by chance so that I can have an idea of what to expect? it would be very helpful to me because afterwards i would do single cell electroporation and it would be a major advantage to know where the cell bodies are..that's why I would try it..
thanks a lot for your help!
Actually i just immersed the fish in hoechst solution for less then one hour and then i rinsed it with water. the fish nuclei were heavily stained.
Sorry that I can't be more helpful...
Any paper or picture about it? Well..i assume that if you stained the fish by immersion i will be able to stain the neurons by dropping some hoechst on the exposed brain in vivo..
Is there any reason why you are not expressing a transgenic construct of GFP that localizes to the nucleus in Drosophila neurons?
I read through the other posts and I see that you wanted to do these on bees. That obviously rules out my answer. Sorry for the confusion.
Hey.. Have you fugured about if Hoeschst works on Bees? I am thinking of whole mounts..will that work?
Hello! I want do live imaging whole drosophila brains for 15hours, with Hoechst to the nuclei but my lines also have gfp and rfp (but they mark only proteins so I'm thinking of using this dna marker to see which cells they really are..). Did you succed previously? Thanks
To me it did not work for live imaging. However to know which cells they are you should not use a generic nuclear marker, because all cells will be positive to it.
I know but through size and my other markers on gfp and rfp I can figure it out. but the Hoechst make the work so much easier. Thanks anyway!
Hey Anat,
I'm also trying to stain the nuclei of living zebrafish larvae but have an issue with proper labeling of all nuclei. How do you treated the larvae within the 1 hour? Which concentration did you use? Thanks in advance.
Pascal
Hello! May I ask you in which dpf did you stain your zebrafish larvae and which concentration did you use? Did it work well finally? Pascal Basdorf
Hello Angelika, I used zebrafish larva 2-4 dpf and used a 1:10000 hoechst stain dilution. It worked well after incubating for 2 h.
Hey Pascal and Anat, thanks for sharing your knowledge. Did you buy any chance already publish these experiments so one could take a look at the protocol or would you be willing to share your Hoechst staining protocol. I would of course acknowledge you with a citation :) I am working with both 2 dpf and and 4-5 dpf old zebrafish
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes