[Histonet] RE: Saponin

[Sharon.Davis-Devine]

Why use Saponin when you can get a commercially prepared solution such as Lysing 1000, which was specifically designed to remove rbc's from Cytology specimens.  Lysing 1000 not only lyses the rbc's but gives you exceptionally preserved cellular material which is amiable with IHC staining. Lysing 1000 is available from Obiter Research, LLC (www.obires.com) and only costs $25 per gallon. We have used it on all of our non-Gyn and FNA specimens for over 12 years now. We use it instead of 95% alcohol for our FNA smears. We prepare a smear, air dry one for a diff quik stain and immediately immerse the other on in a Coplin jar of Lysing 1000.  It removes the rbc's while immediately fixing the cells present on the slide. I recommend using plus slides so the cells don't fall off.  We then rinse the needle in a container of Lysing 1000 which lyses the rbc's and leaves you with a very cellular specimen for a cell block.  We have valildated it's use with IHC staining using our Ventana Benchmarks and seem to get consistently excellent staining results on our cell blocks. I highly recommend it over Carnoys and Saponin.  Let me know if you have any questions.

Sharon Davis-Devine, CT (ASCP)
Cytology-Histology  Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tench, Bill
Sent: Monday, October 25, 2010 10:56 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Saponin

Lots of blood can be a problem in cyto specimens, especially smears.  If
you are making smears from fresh specimens in particular you may elute
the obscuring hemoglobin by dipping the smear directly in acid alcohol
(i think 5% hcl-95% etoh).  We do this frequently for CT guided FNA's
(particularly of the liver and thyroid, which tend to be bloody).  We
also do this "post facto" on those smears even when they have already
been stained with H and E.  The red cell stroma disappears into the
background).  Just lift the coverslip and back up to 95%, use acid
alcohol ( i think it is sold on the commercial market as
"differentiating agent"), and then start staining process all over.
With fresh specimens you can see the hemoglobin elute from the surface
while you dip the slide.  We typically go back into regular 95% Etoh
just to get rid of the acid background (effects the blueing down the
line).
 
Bill Tench
Associate Dir. Laboratory Services
Chief, Cytology Services
Palomar Medical Center
555 E. Valley Parkway
Escondido, California  92025
Bill.Tench <@t> pph.org
Voice: 760- 739-3037
Fax: 760-739-2604
 

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