[Histonet] Histonet Digest, Vol 201, Issue 3 Update info. on old topic?

Steve McClain SteveM at mcclainlab.com
Wed Aug 5 22:41:24 CDT 2020


Tim, Paula and Jeanine
I have little experience w slide scanning, yet I have captured nearly 1.8M slide images, mostly on film coverslipped slides. One year ago we switched back to glass.
1) I suspect what Tim describes may be due to the fact that the refractive index of film differs from glass.
We used film for nearly 20 years and for 4  years I could never get a sharp high power image. This was most noticeable when using 40x lens designed for glass cover slips.
About 16 years ago, we found an Olympus 40x lens w a correction collar allowing for adjustment/ focusing suitable for coverslips of different thickness but useful also with either film or glass. The lens was expensive ($3600 if memory serves me).

See https://www.edmundoptics.com/p/olympus-uplxapo-40x-objective-with-correction-collar/43026/

Perhaps the scanner manufacturer has a similar solution?

2) we were especially diligent (mono-maniacal) about using fresh reagents to keep water out of the last clearing reagent step-film has a nasty reputation for delaminating over time and pulling the sections off the glass. Several major academic centers have years and hundreds of thousands of slides now completely useless due to delamination.

3) film has problems with certain specimens routinely (bone and toenails and the thick cornified layer on volar skin) not cover slipping well leaving bubbles and ‘cornflakes’ or brown spots over the tissue. Cornflake artifact also occurs w water in the last reagent and may be seen on obscenely humid days.

4) film scratches readily when you stack slides or file them too tightly. Film Slides can be filed immediately after exam.  Glass generally needs to wait 2 days before filing or one can glue up a brick of slides by filing too soon.

5) Film is faster and dries faster but glass is sharper and produces better images for my purposes (we switched back to glass 1 year ago- and the Leica model works perfectly well)

6) our old SCA film coverlipper generated more fumes than the new Leica. However all new instruments produce less fumes than older models.

7) glass coverslips are better stored in a desiccator/dry environment, before use.

8) This will sound stupid, yet not all coverslips of clean.- most (3/4) cheap coverslip glass is dirty and cannot be used in our lab where most/every slide is imaged. To determine this pick up an entire stack of coverslips and look through them. If they look clear-good to go; if they appear cloudy, find a new supplier.

9) glass coverslippers are finickyabout viscosity and volume of the mounting media. Once dialed in, we didn’t change media and use one brand and only one.

10) Not all film coverslip rolls are satisfactory either. We had the best luck w Brand S film and did not switch.

Hope these observations help.
Steve
Steve A. McClain, MD
631-361-4000  Cell 631-926-3655

Good morning!

I need to provide some information to my team about film vs. glass coverslipping. I have a lot of positive info. regarding film but are any cons? And I need to come up wit the pros of glass over film. Anyone with extended experience have some current information for me?


Message: 3
Date: Wed, 5 Aug 2020 13:00:37
I am interested in this too, with my main question being the quality of digital images from film coverslipped slides.
Paula Keene Pierce, BS, HTL
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Message: 6
Date: Wed, 5 Aug 2020 15:38:14 +0000
From: "Morken, Timothy" <Timothy.Morken at ucsf.edu>
To: "Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP)" <jqb7 at cdc.gov>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Update info. on old topic?
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Hi Jeanine!

We are using film coverslipping exclusively now and scanning all slides. We moved to film because it dries almost instantly and can be scanned right away. It has worked very well. The only issues, and not specific to film coverslipping,  are that some slides with low contrast sometimes are out of focus. That is primarily IHC slides with low contrast counter stain and low contrast specific staining.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

-----Original Message-----
From: Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP) via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Wednesday, August 05, 2020 5:40 AM
To: 'histonet at lists.utsouthwestern.edu' <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Update info. on old topic?

Good morning!

I need to provide some information to my team about film vs. glass coverslipping. I have a lot of positive info. regarding film but are any cons? And I need to come up wit the pros of glass over film. Anyone with extended experience have some current information for me?

Thanks very much,

Jeanine Sanders, BS,


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