That step is known as differentiation. Clearing involves an organic solvent to
prepare the slides for coverslipping in a synthetic mounting medium.
A regressive H uses differentiation, usually with 1% acid alcohol. A
progressive stain does not. Some companies use a modified progressive using
clarifier which is an acid alcohol using acetic acid.
-Original Message-
From: Anne Murvosh
Sent: Tuesday, October 26, 2021 9:56 AM
To: Mac Donald, Jennifer
Subject: RE: Stain line order
EXTERNAL SENDER- Exercise caution with requests, links, and attachments.
Clear; the tap water makes a blue film on the slides. I think this person used
to use HCL water. I used clarifier 2 in the past. Thanks Anne
-Original Message-
From: Mac Donald, Jennifer
Sent: Tuesday, October 26, 2021 9:49 AM
To: Anne Murvosh
Subject: RE: Stain line order
Clear or differentiate?
-Original Message-
From: Anne Murvosh via Histonet
Sent: Tuesday, October 26, 2021 9:27 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Stain line order
EXTERNAL SENDER- Exercise caution with requests, links, and attachments.
When doing frozen section staining for Mohs we have DI water and don't need a
clearing agent. I am helping set up someone who is using tap water and needs to
clear it. They mentioned that they used to clear first, rinse, then use bluing.
I thought it was the opposite after heme. Rinse, bluing, rinse, clearing. I
don't remember which is first. Actually do you need a bluing if using tap water
since that blues better then DI water? Thanks for your input, it's been 10
years since I had to think about this. Anne
Anne Murvosh
Histology Technician
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