When you tried decalcification in the microwave did you use EDTA?  At what temperature?

Mary Reeves
Technical Specialist
Histology
1-352-265-0680 ext 7-2113

>>> "Gudrun Lang"  10/31/03 06:02AM >>>
We do the same thing with our bone marrows like Loralee, but do not use any
microwave decalcification afterwards.
Our samples are at least four days in EDTA and than are processed in the VIP
like all other biopsies. It works fine.
Our tests with decalcification with mirowave instead of slowly EDTA were not
satisfied.

Gudrun Lang
General Hospital Linz, Austria

----- Original Message -----
From: "Gehan, Loralee" 
To: ; "Gehan, Loralee"
; "'Mary Reeves'" 
Cc: ; "Katherine Raker"

Sent: Thursday, October 30, 2003 9:30 PM
Subject: RE: [Histonet] RE: Microwave Tissue Processors


> We have the RHS-1 also.  What we normally do for our decals is to let them
> sit in EDTA for about a week sometimes longer depending on the amount of
> things that we have to cut/process, etc.  Then we run it on the decal
cycle
> in fresh EDTA at 37C for about 5 hours then process them the next day.  We
> have been getting beautiful results.
> Do you do your entire processing schedule under vacuum?  Or just the
> paraffin?
>
> Loralee
>
> > ----------
> > From: Mary Reeves
> > Sent: Thursday, October 30, 2003 11:13 AM
> > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu 
> > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker
> > Subject: RE: [Histonet] RE: Microwave Tissue Processors
> >
> > Loralee,
> > Which Hacker microwave do you have?  We use the RHS-1 to process our
> > decals.  We are processing with vacuum, this allows us to process at
lower
> > temperatures and still have decreased processing times.  We plan to
start
> > decalcification in the microwave soon.  Any suggestions?
> >
> > Mary Reeves
> > Technical Specialist
> > Histology
> > 1-352-265-0680 ext 7-2113
> >
> > >>> "Gehan, Loralee"  10/30/03 08:08AM
> > >>>
> > I work in an orthopaedics research lab as well and process many many
many
> > mouse hind limbs, calvaria, etc.  We use a microwave processor from
Hacker
> > (milestone medical).  It has decreased our time to process.  We had some
> > trouble at the start with some of our enzyme histochemistry.
> > The trick to the machine is the heat.  It was heating the samples up so
> > much
> > that one of our stains wasn't working.  After much testing we figured
out
> > that all you have to do is decrease the temperature of the processing
and
> > increase the time.  It was basically trial and error because the company
> > markets these for rapid processing for surgical biopsies.
> > We decal on the machine and we also do antigen retrieval and found that
> > most
> > of our antibodies worked well with it.   We are still testing out the
> > capabilities of the machine.  But it has helped this lab become more
> > efficient.
> > Hope that helps.
> >
> > Loralee Gehan
> > Orthopaedics Research Lab
> > University of Rochester
> > > ----------
> > > From: Chan Wai Kam
> > > Sent: Wednesday, October 29, 2003 10:02 PM
> > > To: Mary Reeves
> > > Cc: HistoNet Server
> > > Subject: RE: [Histonet] RE: Microwave Tissue Processors
> > >
> > > Hi Mary,
> > >
> > > I was about to post a question to Histonet about the use of microwave
> > > for processing of tissues when I came across your message below. I
hope
> > > to get some advice.
> > >
> > > I'm from Orthopaedics (research) and I process bone and cartilage
> > > specimens the usual way through fixation in formalin, then
> > > decalcification and so on.  I'm just wondering whether I can use the
> > > microwave to speed up the processing without affecting the quality of
> > > the specimens. Our usual processing for bone takes around 3 weeks
until
> > > embedding so it would be great if we could have something that can
speed
> > > up the process without sacrifing quality.
> > >
> > > Would appreciate any advice out there.
> > >
> > > Thanks
> > > Julee Chan
> > > Orthopaedic Surgery
> > > National University of Singapore
> > >
> > >
> > >
> > > -----Original Message-----
> > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] 
> > > Sent: Wednesday, October 29, 2003 11:38 PM
> > > To: ASelf@gmhsc.com 
> > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker
> > > Subject: RE: [Histonet] RE: Microwave Tissue Processors
> > >
> > >
> > > Amy,
> > > Do you currently have a microwave tissue processor?  We use the
> > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The
times
> > > that you see only represent the time at temperature, they do not
include
> > > the "ramp" up time (the time needed to bring the solution to the
> > > appropriate processing temperature).
> > >
> > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in
> > > Alcoholic  Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced
by
> > > Hacker. Isopropanol can be used instead, however, you may need to
extend
> > > the time at temperature. - 1 hour and 37 minutes at 68 C
> > > Vaporization (Vacuum Drying) -Pressure at 500 mbar
> > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar
> > >
> > > This program takes approximately 4 hours to run, it replaces our 16
hour
> > > overnight program that we previously used to process fatty tissue.
The
> > > only fatty tissue we do not process in the microwave is lipoma.
> > >
> > > Mary Reeves
> > > Technical Specialist
> > > Histology
> > > 1-352-265-0680 ext 7-2113
> > >
> > > >>> Amy Self  10/28/03 01:58PM >>>
> > >
> > > Mary,
> > >
> > > Could you share with me your procedure for processing fatty
> > > tissue in the microwave.
> > >
> > > Thanks,
> > > Amy Self
> > >
> > > -----Original Message-----
> > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] 
> > > Sent: Tuesday, October 28, 2003 1:38 PM
> > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org 
> > > Subject: RE: [Histonet] RE: Microwave Tissue Processors
> > >
> > >
> > > We microwave process all of our biopsy specimens and our fatty tissue
> > > (except lipoma).
> > >
> > > Mary Reeves
> > > Technical Specialist
> > > Histology
> > > 1-352-265-0680 ext 7-2113
> > >
> > > >>> "Hallada, Teri"  10/28/03 11:42AM >>>
> > > I was wondering if anyone out there is using a microwave tissue
> > > processor for routine hospital tissues. Are there any regulations
> > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT
> > > (ASCP) thallada@noch.org 
> > >
> > >
> > >
> > >
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