[Histonet] Histonet Digest, Vol 170, Issue 13 histology hacks

[Seeley, Heather]

Hey Everyone!!

I haven't read this book yet, but we are planning to buy it for our department. I can see that a lot of remarks and reviews are being made and I wanted to give some points from someone who was coming into the field from having a B.S. and several years of training as a lab assistant. I have now been in histology for 11 years, but have only been certified for 6.

1. Congratulations on the book! I think that it is something that will be useful for new techs. 

2. Hack is a word that is generally known in the younger generations as a way to fix things or make things that you wouldn't normally think of. There are many labs out there (I have worked at two) where the physicians and/or pathology administration wouldn't do anything to have the tissue processed in a better way.  
    For example: Fatty tissue being cut in WAY too thick. Things being crammed into the cassettes so it was difficult to fit it into the mold. Staples, sutures, etc. not being removed from the tissue during grossing.  
As a tech, there is not a whole lot that can be done from our perspective to get things changed. We can mention it, but you won't always be in a situation where those changes will be made. (We lived by the motto "Crap in, crap out", but you like to be able to do the best work possible, especially since we are dealing with patient tissue.  This is where techniques would be helpful to employees-so that you can do the best work possible with what you are provided. We used to cut most of our fatty blocks in cold water and lay the slide gently on the water bath to remove the wrinkles. New techs are not going to know these tricks and let's face it, most of the people who know these tricks/hacks are about to retire. So let's not judge or be critical of someone who is trying to make the field more user-friendly. 

Side note: Thank-you everyone who asks and responds to questions on here! I have learned a lot and I think that it is important for us to keep an open dialogue to learn from one another! 


HEATHER SEELEY, HT(ASCP)


________________________________________
From: Steve McClain [SteveM at mcclainlab.com]
Sent: Tuesday, January 16, 2018 7:24 AM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Histonet Digest, Vol 170, Issue 13 histology hacks

I purchased the book and applaud the effort because there is some decent information.  However, the term hack is a poor choice for histology and many of the fixes or secrets described are because some hack failed to do her/his job at an earlier step in the process. (Definition of hack. transitive verb. 1 a : to cut or sever with repeated irregular or unskillful blows. b : to cut or shape by or as if by crude or ruthless strokes. As a noun it is used to mean a mediocre performer or worker; tiresome drudge.)

Some methods, while useful in some settings, have important cons not listed, cons which may be counterproductive. For example using Mercurochrome or Eosin to mark tissue may preclude further testing with fluorescent endpoints, such as FISH.  Plus if you really want to use Eosin to mark the dermis, it is far easier to add used Eosin to one alcohol in the tissue processor. That gives a visible indicator of carryover, indicating need to change or rotate solutions.

Other methods seem (to me) like workarounds or Band-Aids for Labs w poor grossing, poor processing or poor reagents or poor technique or poor method choices, eg, 2.14 describes a situation where an incompetent grosser truly hacks or crudely cuts into unfixed tissue yielding too thick a slice. The real solution is to fix the tissue before slicing. Poor fixation results in poor processing and poor sectioning and poor staining reactions.

For another example, Cassette sponges offer few advantages, while folding lens paper allows the grossers to see through the paper and know all pieces are inside before closing the cassette lid.  The tissue does not stick to it, and small flakes can be scraped from the lens paper at embedding. Last during folding, the forceps can be cleaned at grossing and during unfolding, forceps may be cleaned w the paper after embedding.  Sponges also result in greater solution carryover.

Several colloquial naming conventions, eg, chamber saver 2.16 for underprocessed tissue may be memorable to some readers, yet seem  are odd to me.

This 2.16 method is an especially useful technique which may also be done to extend paraffin time, whenever poor sectioning due to poor processing is encountered at the microtome.
Variation 1 Place the block back into the proper sized mold and return to the heated side of the embedding center for an hour to extend processing (reprocessing). Then remove the old paraffin from the mold w a plastic pipette,  then re-embed, replacing the paraffin w new.
Variation 2 for outside blocks we routinely replace an unknown paraffin from another lab by melting in a mold, and ‘reprocess’ in our (blue ribbon) paraffin for 1 hr in a mold in the embedding center then re-embed.

Good first effort, yet this book could be improved by a good editor, by more collaborators, by illustrations, and the addition of variations or other uses as described above for 2.16.
The font size and format will cause many readers to suffer because of the small font.

Steve
Steve A. McClain, MD