[Histonet] Tissue Microarrays and peroxidase staining artifacts

[Carrie Kyle-Byrne]

Thom,

i beg to differ with your suggestion that the "halo" staining effect is
caused by adhesive in the water bath.  we use only distilled water in our
baths and only plus slides (no additional adhesives) and we still get this
effect when staining on an automated stainer.  my personal observation has
been that this effect is a direct result of the reagent pool receding to the
edge of the tissue.  i say this for two reasons: 1.  i've witnessed the
problem with reagent spreading on plus slides on our dako autostainer (even
though we use 0.1% tween in our buffer), and 2. we also use the shandon
coverplates for manual staining and we never get the "halo" effect on these
slides.

just my two cents,
Carrie Kyle-Byrne, BHS, HT(ASCP)
Assoc. Research Scientist II
Antibody Core Lab
Signal Transduction Research

Exelixis, Inc.
170 Harbor Way
P.O. Box 511
South San Francisco
CA 94083-0511 USA

Phone: (1 650) 837-8023
Fax: (1 650) 837-7220
Email: ckbyrne <@t> exelixis.com

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  ----- Original Message -----
  From: Thom Jensen
  To: histonet <@t> lists.utsouthwestern.edu
  Sent: Friday, October 03, 2003 12:54 PM
  Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts


  Hey Histo World,

  Some people have said that when staining immunoperoxidase using the .6mm
array cores, they get dark stained artifacts around the outer surface of
each specimen.   This also occurs on small biopsy as well.  In our lab we
have found this happens when using a type of adhesive in the water bath to
stick the tissue specimens to the slide.  We have eliminated most of this
problem by using distilled water in our water baths and only plus charged
slides for all our immuno staining.  Every once in a while adhesive may be
needed like for bloody specimens or for bone marrows.  At least that is what
we have found in our lab.
  Any one else have success with other methods of reducing artifacts aournd
or in small specimens?



  For more information on tissue array construction visit:
www.arrayworkshop.com




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