I'm staining fish blood smears and searching for micronuclei. I have seen methods that have only used a 6% Giesma stain solution in Sorenson's buffer (Nwani et al. 2011). I've also seen slides being stained initially with a May-Grunwald Stain and followed by a Giesma stain solution, but I was wondering if it was necessary to stain initially with May Grunwald Stain. I've already stained my slides with a 6% giesma in Sorenson buffer solution at a pH of 6.9 for 45 minutes, however I'm struggling to see the micronuclei. Additionally, if the May-Grunwald stain is needed, can I put my slides back in methenol to strip the stain from the cells and start over? Any insights on my methods would be greatly appreciated. Thanks!
Dear Jillian,
naturally "the experts" will have no problem to by intelligence to the when reading the additional explanation of this your request.
BUT: since the "search for , or , and 'full text search' " is a powerful option in ResearchGate I should greatly appreciate your kind help in
CORRECTING your Q-TEXT with regard to the wrong spelling of >Giesma< which instead has to be : GIEMSA STAIN.
You can do that easily by clicking to the "EDIT" arrow right to the Q-form...go into your text, correct and finally (don't forget!) "SAVE" again (bottom of Q-form then)....
Also, for all those technique-oriented freaks and method-interested research friends: would you mind to at least cite also (a portion) of the TITLE of the reference you mentioned: since naturally one CAN find that reference but perhaps not conveniently....I guess... Thank you for your efforts in the name of "research made easy", best of luck and best wishes, WM
PS: by the way (:-)) and not to tease you: also could be corrected to | methanol |,
(conceding that you are not alone with that mixing up of letters inadvertently:
if you "google" for "methenol" , that yields approx. 16,200 results! and search for
"giesma stain" already yields approx. 9000 results...
Dear Jillian:
The method with May Grunwald-Giemsa is very good for stain blood smears of fish. But I was looking for micronuclei in birds and it works very good.
If you used before other stain, you must to make your own protocol for stain again. So, try one by one first. Because you will not to take off all the
dear Jillian,
I am using the Giemsa stain for fish RBC. it works good and you can see the micronuclei clearly. you should find if your toxin causes micronuclei. if the trouble is due to higher stain absorption by the cells you can try a few dips in methanol. good luck
Hello! I use protocol from here https://www.ncbi.nlm.nih.gov/m/pubmed/22771702/ for my zebrafish. Here they use may grunwald, but i also tryed to stain without it, only with gimsa staining after fixation in ethanol. The result is relatively the same. But without may grunwald there were sometimes parts of the slide with deformed or burst cells...
Nice to read about possibilities after so long time...
nevertheless wondering about the practice of 'scientific imprecision' with regard to specific terms of dye and staining procedure.
Best of luck with your staining problem.
Note only for convenience of further interested readers:
Most interestingly, I found a "private ad" (www.kolabtree.com) when linking to the URL: https://www.ncbi.nlm.nih.gov/m/pubmed/22771702/ as given by Aleksandra Livanova above:
the first attempt of "search" via google (browser used = Googlechrome) linked to:
https://www.kolabtree.com/find-an-expert/--sathesh-kumar-annamalai,
the second attempt displayed more than 5 results-pages of mentions and hints only regarding NCBI-Pubmed, etc...
and unrelated inconclusive company homepages....
and my third attempt, using:
https://www.ncbi.nlm.nih.gov/m/pubmed/22771702 similar as before...
4th attempt (browser MOZILLA-Firefox) eventually revealed:
( https__://www.ncbi.nlm.nih.gov/pubmed/22771702:
NB: before pasting the latter URL into your browser, delete the two underline characters in between: https __ and ://... which I had to insert because otherwise the automated action of RG-software links the URL to the article mentioned in RG (see the following):
Article Gamma radiation induced micronuclei and erythrocyte cellular...
which corresponds to:
Aquat Toxicol. 2012 Oct 15;122-123:125-32. doi: 10.1016/j.aquatox.2012.06.001. Epub 2012 Jun 15.
Gamma radiation induced micronuclei and erythrocyte cellular abnormalities in the fish Catla catla.
by Anbumani S 1, Mohankumar MN.
to be found (only Pay-Per-View, 'PURCHASE pdf = US-$ 35,95)'
https://www.sciencedirect.com/science/article/abs/pii/S0166445X12001889?via%3Dihub
At least for me in Austria and with my browser-configuration ( MOZILLA-FIREFOX-Browser ) the URL
https://www.ncbi.nlm.nih.gov/m/pubmed/22771702/
links to another layout I was used to see for NCBI-/-PubMed-results.
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