Dear Mohamed,

guessing you are dealing with resin embedded blocks and their semithin sections (0.25- 1µm) and not "semithin = very thin paraffin- or cryosections ...

It might be necessary to define more precisely...at least in terms of the resin you have embedded your pancreatic islet-containing tissue...and, perhaps,

what the purpose of using Giemsa in your case would be essential (since there are a lot of semithin staining procedures out in the wild which in general consist of alkaline/basic dyes and can easily be applied to semithin resin sections and most interstingly, contain/use also dyes which are essential in Giemsa stain (e.g. Azure II, Methyleneblue, etc.) in calssical Histology-Histochemistry.

Contrary to that a "real" Giemsa Stain will not stain ordinarily on resin sections, at least unless you remove / solve/"etch" the surface of resin sections by use of highly alkaline and corrosive organic solvents like K- or Na- Methylate (saturated ethanolic solution, old-fashioned and somehow complicated making of!). Even if the solving process has been achieved successfully this does not mean that the "Giemsa Stain true" will yield a reliable positive result.

The mentioned techniques are "old-fashioned", have been used extendedly in the 1980ies to overcome the problems of staining with "histological" staining methods (= more/less hydrous acidic dyes and their combinations) and has led some scientists to get desperate sometimes....

Not to be too long here... if you want to know more, please post here your short comment and request.

Best wishes and regards,

Wolfgang

But islets of Langerhans are endocrine and consist of different cell types which of them produces a specific hormone. Conventional (routine) methods cannot show the specific graniles of these cells.you may use Giemsa stain only for demonstration of chromaffin cell granules.But I think it isn't your aim.

I found a paper describing giemsa stain as a differential stain for islets . I actually looking for differential stain to be applied on semithin secions

thanks a lot Dr Wolfgang Muss for your reply

dear Rita

i am asking about this because i found this paper describing giemsa stain for frog islet . i look for protocol of differential stain to be applied on semithin section .

thanks for your reply

Long again.....

@ Mohamed:

Dear Mohamed,

it is not quite difficult to find positive results for your question, but I guess for a successful Google-search you must have the right "search phrase" (see below)... Even in ResearchGate you'll find answers... nevertheless it was some 30 minutes worth to dig for a specific and comprehensive reply (since I personally am really interested in the matter and I have a day of vacation...(:-)) and - by the way - found literature references and documents I wanted to find also) to your question (see below).

@ Rita: dear Rita, unfortunately you are not quite right, since the notion of "giemsa stain is capable of visualizing several types of pancreatic islets" - namely 4 cell types (see Etayo et al, 2000) - is correct.

@ For both of you I would like to add here the following references:

Google search for < giemsa stain as a differential stain for islets > (!!) found (among others): http://www.unboundmedicine.com/medline/citation/3541292/Giemsa_stain_applied_to_deplasticized_sections_to_identify_pancreatic_islet_cells_

Authors: Díaz de Rada O, Sesma P, López J, Vázquez JJ, Ortiz de Zárate A

Source: Stain technology 61:6 1986 Nov pg 367-73; PubMed ID: 3541292

Abstract

A new application of the Giemsa stain to demonstrate endocrine cells in deplasticized sections of Epon embedded material is described. Its application to the pancreas of Rana temporaria is illustrated. The technique does not require postfixation with OsO4 and is easily performed in 30 min. It allows the easy identification of three types of endocrine cells (A, B, and D). A cells, preferentially located at the islet periphery, stain purple-blue. B cells, which occupy the interior of the islet, display a lilac color. D cells give a strong purple color; they are located both in the periphery of the islets and scattered among acinar cells. Positive identification of the cell types was made by immunocytochemistry and electron microscopy.

Notice: 's new Journal Title = .

Most interestingly I have found teh following reference out of the first 10 results:

cf: http://dspace.unav.es/dspace/bitstream/10171/20172/1/40JCEtayo%20et%20al_GenCompEndocrinol00.pdf

= >Characterization of Pancreatic Endocrine Cells of the European Common Frog Rana temporaria> oh, yes, that's perhaps the frog paper, I guess (:-))....

J. C. Etayo, L. M. Montuenga,1 P. Sesma, O. Dı́az de Rada, J. Rovira, and A. C. Villaro, in: General and Comparative Endocrinology 117, 366–380 (2000)

which should be publicly accessible: marvellous images! From the text sections:

I am Ali Noorafshan, send me your e-mail, I will send the Article:

A simple stereological method for estimating the number and the volume of the pancreatic beta [email protected]

Authors: Ali Noorafshan, Leila Hoseini, Saied Karbalay-Doust, Elham Nadimi

Dear Ali,

really an interesting article and approach to measure (and to stain pancreatic Islet cells)...but unfortunately on / in paraffin block slabs and sections. But I guess the staining of BANGLE R Jr. , 1956, article cf. also http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1942651/pdf/amjpathol00690-0186.pdf ) could be applied to resin sections too provided the resin has been etched away as described in the previous post.

Interesting (also due to other reasons) for me also the variable shrinkage of pancreatic tissue after fixation in neutral buffered formalin. Do you have an explanation for that different percentages (or shouldn't I ask such a silly question)??

Unfortunately in the article which is open access to be seen @ http://www.serena.unina.it/index.php/jop/article/view/802/997,

in your Table 2 the section in the most right positioned row : = "Mean final section

thickness (µm)" is not visible. I greatly should appreciate also receiving (if possible and available) a pdf of your article ( to [email protected]), thank you!), interested also in the "simple" stereological method... (reminding me on my thesis, based mostly on the work of the GUNDERSEN group and their predecessors like WEIBEL and others).

Thanks for posting,

best wishes and regards,

Wolfgang