Re: [Histonet] Tissue Microarrays and peroxidase staining artifacts

From:"Pamela Marcum"


I agree with you.  I have had that effect on manual and automated slides
when the reagents begin to recede or even evaporate in either method.  
Many reasons have been given for this effect however, reagents evaporating,
spreading too thin or not enough applied were usually the problem when I
looked at it very carefully.  Since I have not ever used an adhesive in my
water bath that was not ever the problem.  I can see where it would
contribute if the cores are closely placed and the adhesive and water are
allowed to dry without fully draining the slide, causing a thickened layer
to form at the edges.  The adhesive may well stick to the edges regardless
of draining or even have some protein in it from the manufacturing process
that forms a bond.   We have so many ways we can cause ourselves problems
with the best intentions to improve our work.  

Pam Marcum

> [Original Message]
> From: Carrie Kyle-Byrne 
> To: Thom Jensen 
> Cc: Histonet 
> Date: 10/3/2003 4:34:02 PM
> Subject: Re: [Histonet] Tissue Microarrays and peroxidase staining
> 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
> 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
> 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
> 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:
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>   ----- Original Message -----
>   From: Thom Jensen
>   To:
>   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
> we have found in our lab.
>   Any one else have success with other methods of reducing artifacts
> or in small specimens?
>   For more information on tissue array construction visit:
> --
>   High-speed Internet access as low as $29.95/month*. Click here.
>   *Depending on the local service providers in your area.
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