[Histonet] Ada's Mail Stain line order

[Dee Wolfe]

Hi Anne,

Are they using the term "clearing" loosely, but really mean "rinsing"? The term "clearing" typically refers to the step after the alcohol (dehydration) and before coverslipping (if using a permanent, solvent based mounting media). Not sure why someone would "clear" before the rinse and bluing step in frozen section staining.

For paraffin embedded specimens, the slide would need to Deparaffinize (in clearing agent-xylene or xylene substitute), dehydrate (graded alcohols—high to low), water rinse (preferably DI), stain the nuclei (hematoxylin), water rinse, decolorize (acid alcohol-if staining regressively), water rinse(s), bluing, water rinses, graded alcohol–low to high, cytoplasmic staining (Eosin), alcohol rises, clearing.

For frozen section: 100% alcohol (or other fixative), water rise, hematoxylin (usually progressive, so no need to decolorize), bluing, water rinse, etc…

You may choose to skip bluing as tap water can do it. Although for consistency it should be kept in (tap water pH can with seasons, water treatment facilty processes, etc.)

Dee Wolfe


On Oct 26, 2021, at 12:26 PM, Anne Murvosh via Histonet wrote:

> 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
> 
> 
> 
> This electronic transmission and any documents accompanying this electronic transmission may contain information that is confidential and/or legally privileged. The information is intended only for the use of the individual or entity named above. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on or regarding the contents of this electronically transmitted information is strictly prohibited. If you have received this e-mail in error, please notify the sender and delete this message immediately.
> 
> HIPAA Confidentiality Notice: The information and documents accompanying this e-mail may contain confidential information that is legally privileged and protected by federal and state law. The information is intended for use only by the entity or individual to whom it is addressed, the authorized recipient. The authorized recipient is obligated to maintain the information in a safe, secure, and confidential manner. The authorized recipient is prohibited from using the information for purposes other than intended, prohibited from disclosing the information to any other party unless required to do so by law or regulation, and is required to destroy the information after its stated need has been fulfilled. If you are in possession of this information, and are not the intended recipient, you are hereby notified that any improper disclosure, copying, or distribution of the information is strictly prohibited. Please notify the owner of the information immediately and arrange for its return
> or destruction.
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet