Go for it! start of with 10 sec in Gill-I or Gill-II Hx  which should be great and long enough. If the nuclear detail are not dark enough than add the Hx for another 5 sec.. The only issue you should be careful of - is to be careful that you do not place the slides in Acid/alcohol after that (as you would do with with normal H&E). Just go from Hx to tap water for 5-10 minutes then an appropriately mount.

hope that helps.

Dear Nasibah,

Gill hematoxylin should be ok, I guess. Let me get back to you in a day.

Best,

Mannan 

 This discussion thread from IHC World may help in differentiating:

Histology FAQ

Staining, Histochemistry and Histotechnology

(Frequently Asked Questions)

Dr. John A. Kiernan

Department of Anatomy and Cell Biology

The University of Western Ontario

London, Canada

Mayer's and Gill's hematoxylins

Question.

I would like to know the differences between two types of hematoxylin: Mayer's and Gill's.

Answer 1.

Haematoxylin dye concentration for Mayer is 1 gm/L compared with 2 gm/L for Gill-I. The preservative for Mayer's is chloral hydrate and for Gill it is ethylene glycol. The acidifying agent for Mayer's is citric acid, whereas for Gill it is acetic acid.

Both have very good shelf lives of two years or more under correct storage conditions. They both are used mainly as progressive stains, and are well suited to use as counterstains as well. Gill-I has some some strong adherents for progressive cytology staining.

It is possible to make either of these in a non-toxic formulation without compromising performance or shelf life.

Mike Rentsch

(ausbio[AT]nex.com.au)

Answer 2.

Both stains are hemalums: they are solutions containing hematein (from oxidized hematoxylin), an aluminium salt (the "mordant," which forms dye-metal complexes with hematein), an organic acid to adjust the pH, and a hydrophilic compound (glycerol, ethylene glycol or chloral hydrate). The last ingredient is variously said to modify the solubilities of other ingredients, retard the oxidation of hematoxylin, "preserve" the solution or do nothing at all. In most hemalums the hematein is generated by adding enough of an oxidizing agent (most often the iodate ion) to oxidize about half the hematoxylin. The unoxidized hematoxylin provides a reservoir from which more hematein is slowly produced by atmospheric oxidation. This compensates for the atmospheric over-oxidation of hematein to trioxyhematein (which is useless), thereby prolonging the life of the solution.

The compositions of Mayer's and Gill's hematoxylins are set out below. Mayer's recipe was published in 1863, that of Gill, Frost and Miller in 1974. Gill's hematoxylin closely resembles "haematal-16," a mixture published by J. R. Baker in 1962 that contained ethylene glycol but no organic acid.

MAYER GILL

Hematoxylin 1 g Hematoxylin 2 g

Potassium alum 50 g (0.09M ) Al sulfate 17.6 g (0.03M)

Sodium iodate 0.2 g Sodium iodate 0.2 g

Citric acid 1 g Acetic acid 40 ml

Chloral hydrate 50 g Ethylene glycol 250 ml

Water to make 1000 ml Water to make 1000 ml

Molar ratio of Al ions to haematein molecules in the freshly made solution:

Mayer: 32

Gill: 11

A high ratio of aluminium:dye slows down staining and increases the selectivity for nuclei. Both these hemalums are used progressively; in principle, Gill's should stain more quickly than Mayer's. The effect of excess aluminium is seen most strikingly with Ehrlich's hematoxylin, which is saturated with alum and relies on atmospheric oxidation (slow) to provide a low concentration of hematein from an initially large (6 to 7 g/L) reservoir of hematoxylin. Ehrlich's hematoxylin is the slowest of the progressive hemalum stains (up to 30 minutes, compared with 3 to 10 minutes for Mayer's or Gill's). Hemalums for regressive nuclear staining (e.g. Delafield's, Harris's) have lower aluminium:dye ratios than the progressive stains. Acid-alcohol extracts the dye-metal complex more slowly from nuclei than from other components of tissues.

Some references. These are for practical, rather than chemical or theoretical (i.e. speculative) aspects of hemalum staining.

Baker, J.R. (1962). Experiments on the action of mordants. 2. Aluminium-haematein. Quarterly Journal of Microscopical Science 103: 493-517.

Bancroft, J.D. & Cook, H.C. (1984). Manual of Histological Techniques. Edinburgh: Churchill-Livingstone.

Bancroft, J.D. & Stevens, A., eds. (1996). Theory and Practice of Histological Techniques, 4th ed. London: Churchill-Livingstone.

Ehrlich, P. (1886). Die von mir herruhrende Hamatoxylinlosung. Zeitschrift fr wissenschaftliche Mikroskopie 3: 150.

Kiernan, J.A. (1999). Histological and Histochemical Methods: Theory and Practice, 3rd ed. Oxford: Butterworth-Heinemann.

Llewellyn, Bryan. Stains File.

J. A. Kiernan

Department of Anatomy,

The University of Western Ontario,

LONDON, Canada N6A 5C1

(kiernan[AT]uwo.ca)

Go for it! start of with 10 sec in Gill-I or Gill-II Hx  which should be great and long enough. If the nuclear detail are not dark enough than add the Hx for another 5 sec.. The only issue you should be careful of - is to be careful that you do not place the slides in Acid/alcohol after that (as you would do with with normal H&E). Just go from Hx to tap water for 5-10 minutes then an appropriately mount.

hope that helps.

I used to do Oil Red O staining and hematoxylin gives great results as a counterstain that stains the nuclei with blue to differentiate them from the lipid droplets! my paper is attached

Article GCN2 is required to increase fibroblast growth factor 21 and...

you need a nuclear stain to identify cell population as oil red identify oil drops in the cell cytoplasm. I agree with the use of hematoxylin but an easy fast alternative is toluidine blue. There are several free publications and protocols easy to find in the internet

Honestly:  cf. two replies in Researchgate thread:  @  https://www.researchgate.net/post/Does_anyone_have_a_protocol_for_counterstaining_an_Oil_Red_O_stained_slide_with_H_E ; (maybe you might use the SEARCH-QUESTION  function in ResearchGate for a text search | ORO   Oil Red O   counterstain  | you may perhaps be surprised about similar questions and their answers....(:-))

also, concerning literature/research articles:  cf. 

CHOLEWIAK, R. W., L. BUTCHER AND N. M. KETTLEWELL. Oil red O and

[NB: HARRIS's]

hematoxylin: a rapid histologic technic. PHYSIOL. BEHAV. 3 (4) 585--586, 1968  cf.@ https://pdfs.semanticscholar.org/e63e/74e66b856312eb68a14393f06038944ad370.pdf ; 

- For staining adipocyte cultures: ORO and HE [also HARRIS's],  including procedure(s) step by step and chemical product sources....:  cf.@: http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_ManualsProductInstructions_Oil_Red_O_Stain_for_In_Vitro_Adipogenesis.pdf 

- ORO - For the histological visualization of Fat Cells and Neutral Fat, for fresh or frozen tissue sections:  using modified Mayer's hematoxylin,  cf. @

http://www.abcam.com/ps/products/150/ab150678/documents/ab150678-Oil%20Red%20O%20Stain%20Kit%20(website).pdf

- Masserdotti et al,2006: Use of Oil Red O stain in the cytologic diagnosis of canine liposarcoma ,  look into the Preview -page 37:  Mayer's hematoxylin; cf @:   http://onlinelibrary.wiley.com/doi/10.1111/j.1939-165X.2006.tb00086.x/abstract  

- "Best Ever Oil Red O Protocol" (blog in HISTONET by Donna Emge, 2011), using HARRIS's hematoxylin :                      cf @:  http://lists.utsouthwestern.edu/pipermail/histonet/2011-January/054551.html

- Protocols for Adipogenic Differentiation Assays for Characterization of

Adipose Stromal Cells (ASC), Hematoxylin (not specified, from VECTOR)

cf. @ https://catalog.coriell.org/0/pdf/1401_38.pdf

Disclaimer: not affiliated with any of the mentioned companies, no financial interest... This post only for convenience to compare several protocols -how staining was done including different tasks and processing conditions.            Best of luck! 

Dear everyone..Thank you very much. Your replies greatly appreciated. It works with Gill.  Thank you for all literature given. It really made me understand the concept.

Is there a specific mounting medium that should be used when doing ORO and hematoxylin staining?

Sara: You should use an aqueous mounting media as you can't use alcohol or xylene to clear your slides.