[Histonet] Biopsy Run

[Hugh Luk]

Hi again Kim,

>From your previous emails, it seems to be something your tissue processors cannot handle.  Review your equipment and consumables.  Below, is (almost) everything I think about when I hear "Some" tissues are staining/processing" irregularly:

Formalin?  Formalin-substitute?  Recycled?  Who makes your formalin?  Some inconsistencies in vendor bulk formalin cubes have been noted.

Processors?  You said you have two.  What types or brands?  Dip-and-dunk (open) or infiltration chamber (closed)?  If closed-type, when did your folks last clean or replace the tubing?  Retort gasket?
Cassette-type?  Can your cassettes "Float" in the processor?  Biopsy sponges used?  http://lists.utsouthwestern.edu/mailman/htdig/histonet/2004-April/004782.html?
Alcohol?  Check with a Hydrometer?  Who grades your alcohol?  Recycled?
Xylene or xylene-substitute?  Recycled?
Infiltrating Paraffin?  Temperature?  Confirm with a thermometer?

Embedding - Temperature?  Cassettes in holding tank for too long?  

Mictotomy - Soak your blocks in water/ice?

Drying - How long in incubator?  Temperature? 


As for your question about biopsy processing times, refer to Histotechnology A Self-Instructional Text, any edition, by Freida L. Carson, ASCP Press.  In the 2nd edition pp 32-34 (note there is a troubleshooting portion too), Protocol #3 lists basically the processing run that I do overnight (and I think you do to), but I start in formalin.  Everything is listed as 15 minutes, but it will also depend on what types of tissues are defined as "Biopsies," and you should extend the times as needed.  I use 30 minutes, with some longer as listed.  Pressure and vacuum is up to you, but I only use it throughout the paraffin steps.  

10% alcohol
95% alcohol
95% alcohol
100% alcohol
100% alcohol (45  minutes)
Xylene
Xylene (45 minutes)
Paraffin 
Paraffin
Paraffin (45 minutes)

Investigate these options and respond if needed.  

Hope this helps,
Hugh-Hawaii


> Date: Mon, 28 Mar 2011 12:32:32 -0800
> From: kkmarshall <@t> anthc.org
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Biopsy Run
> 
> Hello out there!!!
>   
>     Just a couple of quick questions...I wrote before with a "artifact"
> problem and I thank everyone for their input.  Bad thing is, I am still
> having issues.
>  
>   Would my fellow Histo Techs please share with me the times you use on
> your Biopsy Run's as well maybe give me some advise on biopsy cassettes.
> I have used 2 different kind and just cant help but wonder if air
> getting caught in the cassette may be one of my problems.
>  
>   Thanks in advance for your help...I sure need it.
>  
> Kim
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