If the objects of interest are stained with alcian blue, any pink or red counterstain should be OK. Aluminium-nuclear fasr red if they want to see nuclei; eosin if a general pink to red background is sufficient. H&E is also OK after alcian blue because the two blue colours are clearly different. John Kiernan = = = On 14/12/11, [email protected] wrote:
> > > > > > > > > > > > > > > > > > John, > > > > > > No offence, there is so much in histology I don’t know, and any way I can > learn is good. I use your textbook all the time. > > > In this case I meant to use chlorantine fast red 5B, which they want to use, > instead of nuclear fast red, which is in my protocol. > > > What protocol would you suggest for them? > > > > > > Thanks, > > > iliana > > > > > > > > > > > > > From: John Kiernan [mailto:[email protected]] > Sent: December 14, 2011 2:31 AM > To: Dimitrova, Iliana; [email protected]; > [email protected] > Cc: [email protected] > Subject: RE: [Histonet] Stain recipe. Nuclear fast red > > > > > > > Sorry to make a blunt contradiction, because a few histonetters seem to take > offence very easily. > > > > > > > > Nuclear fast red cannot replace any "chlorantine" dye. The "chlorantine" > dyes in the literature and lore of staining are anionic; they bind to > proteins such as those in cytoplasm, collagen etc. > > > > > > > > The name nuclear fast red (Kernechtrot in German) goes with an anthraquinone > dye long obsolete in the world of textile dyeing. It has a Colour Index (CI) > number - 60760 - but it doesn't have a CI application name. Possibly the only > use of this compound is as a biological stain. It is used as its aluminium > complex, which provides red staining of cell nuclei and weak background pink > in cytoplasms and collagen. The Biological Stain Commission (BSC) certifies > nuclear fast red. > > > > > > > > Vendors of kits that include aluminium-nuclear fast red as a nuclear > counterstain should check that they are selling certified nuclear fast red. > The NFR standards were published in 2007. > > > > > > > > John Kiernan > > > > Anatomy, UWO, London, Canada > > > > (Secretary, Biological Stain Commission) > > > > = = = = > > > On 13/12/11, [email protected] wrote: > > > > Hi Catherine, > > I have staining protocol for alcian blue and nuclear fast red, see bellow. I > looked in Conn's biological stains book (9th edition,p.188)and saw that > chlorantine > fast red 5B is soluble in water. I think you can substitute Nuclear fast red > with chlorantine > fast red 5B and try and see the results. Connective tissue will be deeper > red. Use controls and try it. > > Iliana Dimitrova, MSc, RT > Histology Supervisor > Medical Education and Laboratory Support Services (MELSS) > Faculty of Medicine > Memorial University of Newfoundland > St. John's, NL Canada > > Alcian Blue Staining Protocol > > NovaUltra Special Stain Kits > > Description: Alcian blue stains acid mucosubstances and acetic mucins. > Excessive amounts of non-sulfated acidic mucosubstances are seen in > mesotheliomas, certain amounts occur normally in blood vessel walls but > increase in early lesions of atherosclerosis. Strongly acidic mucosubstances > will be stained blue, nuclei will be stained pink to red, and cytoplasm will > be stained pale pink. > > Fixation: formalin fixed, paraffin embedded tissue sections. > > Solutions and Reagents: > > 3% Acetic Acid Solution: > > Glacial acetic acid ----------------- 3 ml > Distilled water --------------------- 97 ml > > > Alcian Blue Solution (pH 2.5): > > Alcian blue, 8GX -------------------- 1 g > Acetic acid, 3% solution ----------- 100 ml > Mix well and adjust pH to 2.5 using acetic acid. > > > 0.1% Nuclear Fast Red Solution: > > Nuclear fast red ------------------- 0.1 g > Aluminum sulfate------------------ 5 g > Distilled water ---------------------100 ml > Dissolve aluminum sulfate in water. Add nuclear fast red and slowly > heat to boil and cool. > > Filter and add a grain of thymol as a preservative. > > > Procedure: > > 1. Deparaffinize slides and hydrate to distilled water. > 2. Stain in alcian blue solution for 30 minutes. > 3. Wash in running tap water for 2 minutes. > 4. Rinse in distilled water. > 5. Counterstain in nuclear fast red solution for 5 minutes. > 6. Wash in running tap water for 1 minute. > 7. Dehydrate and through 95% alcohol, 2 changes of absolute alcohol, 3 > minutes each. > 8. Clear in xylene or xylene substitute. > 9. Mount with resinous mounting medium. > > Results: > > Strongly acidic sulfated mucosubstances -------------- blue > Nuclei ------------------------------------------------------ pink to > red > Cytoplasm ------------------------------------------------- pale pink > > > Positive Controls: > > Small intestine, appendix, or colon. > > -----Original Message----- > From: [email protected] > [mailto:[email protected]] > <[email protected]]> On Behalf Of Ross, Catherine > Sent: December 13, 2011 5:45 AM > To: [email protected] > Cc: Grainger, Shirley > Subject: [Histonet] Stain recipe > > All, > > Greetings from across the pond! > > One of our histology technicians is looking for a recipe for an alcian > blue/sirius red stain, otherwise known as "Alcian blue 8G with chlorantine > fast red 5B". It is not one I have used but I suggested tapping into the > wealth of knowledge on this forum - can anyone help, please? > > Thank you in advance, > Catherine > > > Dr. Catherine L. Ross DVM MSc(VetPath) MRCVS > Covance Laboratories (UK) Ltd. > Otley Road, > Harrogate, N. Yorks. > HG3 1PY > > T: +44 (0) 1423 848759 > E: [email protected] <mailto:[email protected] > <[email protected]>> > > > > ----------------------------------------------------- > Confidentiality Notice: This e-mail transmission > may contain confidential or legally privileged > information that is intended only for the individual > or entity named in the e-mail address. If you are not > the intended recipient, you are hereby notified that > any disclosure, copying, distribution, or reliance > upon the contents of this e-mail is strictly prohibited. > > If you have received this e-mail transmission in error, > please reply to the sender, so that we can arrange > for proper delivery, and then please delete the message > from your inbox. Thank you. > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This electronic communication is governed by the terms and conditions at > http://www.mun.ca/cc/policies/electronic_communications_disclaimer_2011.php > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > This electronic communication is governed by the terms and conditions at > http://www.mun.ca/cc/policies/electronic_communications_disclaimer_2011.php > > > _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
