[Histonet] Microwave retrieval

mark.lewis <@t> thermo.com mark.lewis <@t> thermo.com
Fri Oct 22 07:46:48 CDT 2004


Patty,

I think you hit the nail on the head when you said "I believe consistency
in what you do is the key to any antigen
retrieval." I believe it is important for all aspects of the Histo lab. It
makes trouble-shooting problems much easier if the work is all done in a
consistent manner.

Have a nice day


Best regards,

Mark

Mark Lewis
Product Specialist
Anatomical Pathology
Clinical Diagnostics
Thermo Electron Corporation
(412) 747-4013
(412) 788-1097
E-mail: mark.lewis <@t> thermo.com



                                                                                                                                                    
                      Patti Loykasek                                                                                                                
                      <ploykasek <@t> phenopath.com>             To:       histonet <histonet <@t> pathology.swmed.edu>                                       
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                      histonet-bounces <@t> lists.utsouth        Subject:  Re: [Histonet] Microwave retrieval                                            
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                      10/21/2004 12:09 PM                                                                                                           
                                                                                                                                                    
                                                                                                                                                    




Ed,
    I've done antigen retrieval many ways, and there are pros & cons to
each. I believe consistency in what you do is the key to any antigen
retrieval. I do not recommend microwaving with open containers - it is far
too easy for there to be hot spots, loss of fluid, etc.... That being said,
I do think it is possible to microwave retrieve in a closed container such
as a microwave pressure cooker. Some of the keys to this technique are:
use
the same volume of liquid, the same # of slides (using blank slides if
necessary) each & every time, monitoring with your microwave the time it
takes to get up to pressure & how long you want your slides to remain under
pressure, careful cooling down of slides. I¹d be happy to provide the
particulars of how we do it, if anyone is interested. In a microwave
pressure cooker the fluid becomes ³super heated² - much hotter than in a
steamer & you have the added pressure. We have found it improves staining
with many difficult targets & nuclear antigens.
I would note that I work at a reference lab, and our tissue is fixed &
processed many different ways.
 If you can control the fixation & processing, and optimize it, I think a
steamer works great & we use it for many antibodies. Again, consistency is
the key. We preheat solutions before adding slides, only adding slides when
the solution has reached 95 degrees C. To monitor the temperature we have
thermometers with slender, metal probes that fit thru the holes in the
steamer ( alternately, I¹ve had my husband carefully enlarge the steamer
lid
holes to accommodate a standard thermometer). After adding slides, we start
our retrieval time when the solution is back up to 95. We do the same
retrieval time & cool down time each time.
Well, I hope this helps you out. Feel free to contact me for more info.
Good
luck with your retrieval & staining.


Patti Loykasek
PhenoPath Laboratories
Seattle, WA




> Can anyone give me the pro's and con's of steamer/pressure (Decloaker)
vs.
> microwave retrieval ?
> Our Doc wants us to change from the Decloaker to microwave. Is a
household
> microwave adequate or do we need a lab grade unit?
> Does the microwave save time and improve turn around time?
>
> Thanks in advance
> Ed Harris  HT ASCP MT ABB
>
>
>
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