How to prepare Rhodamine B for staining protoplast?
How to stain?
What is the excitation level i should use to observe stained protoplast?
Hi,
Rhodamine B is water soluble at 50 g/L.
As it is rather hydrophobic, you should prepare this solution in glass vials to avoid plastic staining with RhB.
As RhB will not stain protoplasts by itself, I guess you want to use Rhodamine B Iso-ThioCyanate (RITC)for membrane protein staining.
If then, add 1/1000 of RITC stock solution to your proto suspension, let stain for 30 minutes, and observe with a fluorescence microscope, using TRITC filter or equivalent (green excitation, red emission).
Cheers,
Oliv
Hello, just have a look also into the recent RG-thread: https://www.researchgate.net/post/Rhodamine_B-isocyanate_staining_of_protoplasts ;
Also:
Article Fluorescent Vital Stains for Complementary Labelling of Prot...
=http_s://www.ncbi.nlm.nih.gov/pubmed/2464211 {NB: to ‚arrive‘ at the real PubMed-page, which I intended to insert here – followed by „automatic“ redirection to an >article profile< in ResGate: before pasting or clicking into the URL, delete the „underscore _“ in between http_s:// !}
or also:
Article Methods of staining and visualization of sphingolipid enrich...
=*) see below‚Methods of staining and visualization of sphingolipid enriched and non-enriched plasma membrane regions of Arabidopsis thaliana with fluorescent dyes and lipid analogues‘ by Blachutzik et al, 2012 in Plant Methods 2012; 8: 28:
*)= : {same applies for this URL as before…(;-() }
http_s://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544639/ and its URL for the open access PDF: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544639/pdf/1746-4811-8-28.pdf {= http_s://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544639/pdf/1746-4811-8-28.pdf}
It would be nice to follow your successful staining by means of a short post about the results (hopefully you can achieve....)(:-))
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining