Re: [Histonet] Microwave Processing

From:Kathleen Spencer

Do you guys know each other? I love this battle of the minds, but who am 
I really to believe?
  You have truly brightened my day and made me laugh when I was blue.

-Kathleen


On Wednesday, April 21, 2004, at 10:51  AM, Kemlo Rogerson wrote:

> Geez, tetchy begger aren't you. I'll answer when I've thunk about it and
> give you an answer that is both lucid and structured.
>
>  this used to be fun; why are some people so......
>
> Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS
> Tel: 0208 970 8414
> Mob: 07830 196072
> Mobile E-Mail kemlorogerson@3mail.com
> FAX & Answer Phone 0871 242 8094
> E-mail Accounts:
>              kemlo@tiscali.co.uk or
>              kemlo1@btinternet.com
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>
>
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>
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall
> Terry Dr,Consultant Histopathologist
> Sent: 21 April 2004 15:51
> To: Kemlo Rogerson; Steven E. Slap; kevin williams;
> histonet@pathology.swmed.edu
> Subject: RE: [Histonet] Microwave Processing
>
> Kemlo asks:
>
> "Am I talking a load of..........."
>
> Inevitably:-)
>
> Kemlo,
>
> You talk as if formalin was the only fixative. What about the group
> called, oddly enough, coagulative fixatives?
> Now .... I wonder how they work. Let me see ......
> Furthermore, you flit between the several different subjects in an
> unstructured way.
>
> When you say :
> "Its job is also that of preventing putrefaction both by endogenous and
> exogenous means
> (lysosomes and bugs)."
>
> You are wrong on 2 counts.
> Its effect may be to prevent putrefaction, but hardly its job. (I
> realise that some books list this, but a dehydrated specimen in wax is
> more at risk from mice than bacteria).
> Furthermore, the action you refer to, attributed to lysosomes, is
> autolysis, not putrefaction.
>
> Furtherfurthermore, a fixed egg will only elude putrefaction by virtue
> of the retained/residual fixative fixing any bacteria with ungodly
> thoughts toward the egg, wouldn't it?
> That is to say, "resistance to putrefaction" will be a property that
> resides in the fixative not in the fixed egg.
>
> Yours lovingly,
>
> Confused from Rotherham
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
>  Consultant Pathologist
>  Rotherham General Hospital
>  South Yorkshire
>  England
>         terry.marshall@rothgen.nhs.uk
>
> -----Original Message-----
> From: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk]
> Sent: 21 April 2004 15:16
> To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap';
> 'kevin williams'; histonet@pathology.swmed.edu
> Subject: RE: [Histonet] Microwave Processing
>
>
> Um....... A boiled egg is stabilized but not fixed; does that help? A
> fixative is designed not only to stabilize proteins by coagulation, but
> in most cases it links chemically to the proteins. Its job is also that
> of preventing putrefaction both by endogenous and exogenous means
> (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you
> fixed it, then it wouldn't; would it? Heat stabilizes by coagulating
> protein doesn't it, formalin forms links between reactive sites on the
> proteins, doesn't it?
>
>
>
> Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS
> Tel: 0208 970 8414
> Mob: 07830 196072
> Mobile E-Mail kemlorogerson@3mail.com
> FAX & Answer Phone 0871 242 8094
> E-mail Accounts:
>              kemlo@tiscali.co.uk or
>              kemlo1@btinternet.com
> Disclaimer: The information contained in this message and/or any
> attachments(s) may be of a private and confidential nature, and is
> intended solely for the attention of the addressee. If you have received
> this message in error or feel you should not have been the intended
> recipient, please return it and any attachments to the sender
> immediately. All messages relating to this communication should then be
> deleted from your system. Unauthorised usage, copying, disclosure or
> alteration of this message and/or attachment(s) is strictly prohibited.
> Barking, Havering and Redbridge Hospitals NHS Trust will not be held
> responsible for any direct or indirect damages which may arise from
> alteration of this message or any attachment(s), by a third party or
> resulting from the transmission of a virus.
>
>
>
>
>
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall
> Terry Dr,Consultant Histopathologist
> Sent: 21 April 2004 12:48
> To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu
> Subject: RE: [Histonet] Microwave Processing
>
> My confusion has not been dispelled.
>
> Steven writes:
>
> "The saline will not work
> alone as a fixation step, except in the case of
> very small biopsies (as written up in an article
> by Tony Leong)- ..."
>
> In the next post, we get:
>
> "Yes, in the Leong method for biopsies, the specimens are heat
> stabilized in the microwave in saline, and not really chemically
> fixed.  They get fixed in the ethanol in a traditional processor ..."
>
> These statements are mutually incompatible.
>
> Moreover, what is the difference between "stabilisation" and "fixation".
>
> A further muddle is introduced by:
>
> "not really chemically fixed."
>
> Well, hell, are they fixed or no? Heat fixation is fixation - who cares
> whether chemically fixed?
>
> Of course, the major problem is that mechanisms of fixation are varied
> (with the fixative) and not well understood.
> (Speak for yourself do I hear someone say?)
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
>  Consultant Pathologist
>  Rotherham General Hospital
>  South Yorkshire
>  England
>         terry.marshall@rothgen.nhs.uk
>
> -----Original Message-----
> From: Steven E. Slap [mailto:siksik03@comcast.net]
> Sent: 17 April 2004 22:54
> To: Marshall Terry Dr, Consultant Histopathologist; kevin williams;
> histonet@pathology.swmed.edu
> Subject: RE: [Histonet] Microwave Processing
>
>
> Hi Terry & HistoNetters
>
> I have done lots of successful large organ
> stabilization in saline at 65C for 20-35 minutes
> (depending on the size of the organ.  However, it
> is important to emphasize that the purpose of
> this step usually is to firm up the tissue so
> that it can be subsequently cut up into thinner
> pieces for processing, which must include a
> fixation step, either in the microwave or on a
> conventional processor, in formalin or some
> non-formalin fixative.  The saline will not work
> alone as a fixation step, except in the case of
> very small biopsies (as written up in an article
> by Tony Leong)-  a procedure I have not tried
> myself.
>
> best regards,
> Steven Slap
> Microwave Consultant
>
>
> At 3:58 PM +0100 4/16/04, Marshall Terry Dr,
> 	Consultant Histopathologist wrote:
>> Sorry to but in - but on a related question -
>> does anyone do microwave fixation in saline as
>> described by for instance the Melbourne group,
>> which involves bringing up to 65C in saline?
>> Have tried it this week and the sections are even crummier than usual.
>> When I was in Tasmania, a local private lab. did
>> it and their sections were fine.
>
>
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