There are papers stating it is possible to stain nucleus but I'm having difficulties. Please help me. Thank you all in advance.
yes the Davidson's fixative is a rapid fixative which gives good nuclear to tissues even with minimal formalin pigment, but my question is did you get the process right and what did you see. these are some of the questions i think we need to answer. you will find this material useful
https://www.researchgate.net/publication/23185805_An_antibody_panel_for_immunohistochemical_analysis_of_the_retina_in_Davidson%27s-fixed_paraffin-embedded_eyes_of_rats
best of luck in your research
Article An antibody panel for immunohistochemical analysis of the re...
these are rat tissues, i was thoroughly impressed with the preservation of details when fixed by davidson's fluid. they were post fixed for 4 days. i stained them for nissl and it turned out to be just fine. although when i stained it synaptophysin and counterstained it with DAPI, i never found the nucleus stained.
Dear Yogesh,
affinity of a classical histochemical dye to "structures contained in cells" (like "Nissl") is considered to be quite different from affinity of (specific or unspecific) Ab's (re. synaptophysin) to "structures & protein molecules contained in cells" .
Also (IMHO), no fixative existing (IMHO) without some adverse effects in preservation of molecules&structures too.
Nevertheless (and you might have seen these already) there are articles available stating that "modified Davidson's fixative" (and perhaps also DAVIDSON’S original formula) is superior in fixation in some tissues, compared to fixation with BF(Bouin's Fluid), or other fixatives, especially in terms of IHC):
cf: Fixation of testes and eyes using a modified Davidson's fluid: comparison with Bouin's fluid and conventional Davidson's fluid, by JR Latendresse, AR Warbrittion, H Jonassen, and DM Creasy in: Toxicol Pathol.2002 Jul-Aug;30(4):524-33, see: http://www.ncbi.nlm.nih.gov/pubmed/12187944 (free full text available, description of the modified Davidson’s fixative as well as methods for antigen Retrieval),
as well as e.g.
An antibody panel for immunohistochemical analysis of the retina in Davidson's-fixed, paraffin-embedded eyes of rats, by: JS McKay, SJ Steele, G Ahmed, E Johnson, K Ratcliffe, in: Exp Toxicol Pathol. 2009 Mar;61(2):91-100. doi: 10.1016/j.etp.2008.06.005. Epub 2008 Aug 20, see: http://www.ncbi.nlm.nih.gov/pubmed/18718747 or (this article/ .pdf MIGHT be available open access also to you at:
http://ac.els-cdn.com/S094029930800105X/1-s2.0-S094029930800105X-main.pdf? (if you can't access, ask me for the pdf),
BUT: the latter paper stating also: [Citation] “Modified Davidson’s solution, while providing an acceptable fixation for retina and cornea, produces inferior cellular preservation compared with Davidson’s fixative. Both of these fixatives also provide relatively poor lens fixation with inconsistent staining of the inner core (Latendresse et al., 2002). In contrast, formalin fixation, preferred for other tissues and also for immunohistochemistry, causes artifactual cellular shrinkage and poor cellular and nuclear preservation in the retina” [End of citation].
You have not indicated your tissue to be examined (only “rat”) but “striatum” in the RG keywords points to , as Matthew IBBS pointed out already.
So this (and from at least these cited articles – and there are many others out in the wild….) might (does) mean that there are differences in the preservation efficiency (at least for IHC-staining application) of the
“original Davidson’s Fixative “ (BTW: interesting “genesis” of getting known as potent fixative for eyes!) as it (the recipe) was published originally by Moore KL, Barr ML (1954). Nuclear morphology, according to sex, in human tissues in Acta Anat 21: 197–208.
and
Moore KL, Graham MA, Barr ML (1953). The detection of chromosomal sex in hermaphrodites from a skin biopsy, in Surg Gynecol Obstet 96: 641–648)
versus
“modified DF” as published by Creasy DM, Jonassen H (1999): Histological fixation of testes: Use of Davidson’s fluid as an alternative to Bouin’s fluid, in Vet Pathol 36: 518.
So you might try and fiddle around with some variations, either with the fixative itself, or the fixation time and further processing, or with antigen retrieval, respectively… unfortunately I don't have experience with this particular (special) fixative
Best wishes and good luck, Wolfgang
thank you Dr. Muss, i wish to experiment around with some variations
You're welcome, dear Dr. Yohgesh K. Sathyamoorthy.
Another article I found yesterday evening (you may have it in your files already) could be a variant to be tested (fixing eyeballs with Davidson's Fix first for 48 hrs, then change to / and kept for 24 hrs , IHC for caspase-1,-3 on paraffin-embedded material after AR (AntigenRetrieval):
M Saenz-de-Viteri, et al, 2014, Oxidative Medicine and Cellular Longevity Volume 2014, Article ID 637137, 10 pages, http://dx.doi.org/10.1155/2014/637137; (C) Hindawi Publishing Corporation
http://www.hindawi.com/journals/omcl/2014/637137/abs/
best regards and good luck, Wolfgang
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