[Histonet] Microwave retrieval

[Patti Loykasek]

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
> 
> 
> 
> This E-mail message (including attachments, if any) is intended for the use of
> the individual or entity to which it is addressed and may contain information
> that is privileged, proprietary, confidential and exempt from disclosure.  If
> you are not the intended recipient, you are notified that any dissemination,
> distribution or copying of this communication is strictly prohibited. If you
> have received this communication in error, please notify the sender and erase
> this E-mail message immediately.
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet