[Histonet] Re: "waviness"

Gayle Callis gcallis <@t> montana.edu
Thu Jul 28 11:32:34 CDT 2005


It may be more than how you pick up from the water bath.

Make sure your disposable blades are sharp, of high quality and maybe 
change more often to a sharp edge.
Make sure the knife angle is correct for the blade you are using.
The sections should come off the block onto blade without compression.
Check all levers, microtome knife holder adjustments have to be tight.

Check you water bath temperature for the paraffin you use,  the sections 
need to flatten out without exploding, and you are picking up nicely - do 
this quickly so section hits slide surface evenly - Marcia from Newcomer 
Supply gave excellent hints on getting sections to the surface so it STICKS 
onto plus charge.   If your waterbath is too cool, the section will not be 
flat at times - a problem I have when I am sectioning too fast and don't 
take the time to wait for sections to flatten - I then up the temp a few 
degrees to accomodate speed coupled with impatience.  Be careful doing the 
latter, you don't want to have sections come apart.

Do not put any adhesives in the waterbath, it should be distilled water (if 
you use tap and water is hard, i.e. mineralized, that will afftect how 
sections sticks) adhesives coat the plus charge and negate the plus 
effect.  This is all given on the package insert inside Plus Charge slide 

When you pick up a section onto Plus slides, drain the section vertically 
so the water drains OFF slide - down, away and out from under 
section.   You are correct in not going to warming platform immediately 
after picking up section as water will pool under the section,  creates 
uneven adherence.

Another thing, additives in paraffins are heavy and can settle to bottom of 
dispenser when paraffin melts. Stir the paraffin before embedding to 
redistribute these additives.  You want the paraffin well mixed to have 
benefits of the additives for  good sectioning.

Good luck

At 05:46 AM 7/28/2005, you wrote:

>One of our pathologists is complaining about having to focus up and down too
>much while reading slides.  I have tried many things to resolve the problem
>but nothing is making him happy.  I have changed the way we pick up from
>water bath to be sure slides are perpendicular (straight up and down) when
>picking up sections, we use only plus slides, I've asked that slides not be
>placed on the warming platform after picking up.  Does anyone have any ideas
>about how I can fix this problem?
>Julie Sanders
>Supervisor, Anatomic Pathology
>Cincinnati VA
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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