RE:MUCICARMINE

Dear Histonetters:
     I have questions regarding a mucicarmine stain being done on distal stomach.
The stain has been done on different sections of stomach, distal to insure mucin, but it is not turning out. We have checked the chemicals, expiration dates (etc.). The same recipe and solutions have worked on other types of tissue. Is there a specific reason why stomach is resistant to mucicarmine? Any suggestions are appreciated.

Thankyou,
Denise Bland-Piontek, HTL(ASCP)
 
>>> HistoNet Server <histonet@pathology.swmed.edu> 03/04/00 11:00PM >>>

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Date: 4 Mar 2000 03:45:17 -0600
From: "Tony Henwood" <henwood@mail.one.net.au>
Subject: Re: 

Dear Marianne,
Would someone tell me, my gold chloride made in triple distilled water
froze, is it still good to use?
 
I can't imagine any staining technique that would be affected.
Both water and gold chloride are unaffected by freezing.
Why do you keep the solution in the fridge?
Wouldn't room temp storage do?

Regards, Tony




Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital
Sydney, AUSTRALIA

http://www2.one.net.au/~henwood 
http://www.pathsearch.com/homepages/TonyHenwood/default.html 


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Date: 4 Mar 2000 09:00:43 -0600
From: "Tim Morken" <timcdc@hotmail.com>
Subject: Re: cryostat decontamination

Susan,

You have discovered what I did last year. I did a literature search and came 
up with nothing. Not one article on cryostat decontamination (at least none 
that had documentation on why a certain method was used).

Here are some methods I found that did not have any documentation to back 
them up as to effectiveness:

In all cases you must defrost the cyrostat before decontaminating.

1) Wash out with 100% alcohol (be careful of flammable fumes)

2) wash with 10% formalin or Cidex (2.5 % glutaraldehyde) followed by water, 
followed by 100% alcohol.

This one is documented, but not published.
3) Put a dish of 37% formalin ("concentrated formalin", "formaldehyde") in 
the defrosted cryostat and leave overnight. This is from an unplublished 
study by a cryostat manufacturer and is the method recommended by Shandon 
for their cryostats.

Textbooks will say to "decontaminate the cryostat" but either do not give 
any instructions on how to do so, or the instructions they give are not 
backed up with studies documenting effectiveness.

No cryostat manufacturer, except Shandon, gives instructions on how to 
decontaminate a cryostat.

I am working on a study to determine the need and effectiveness of cryostat 
decontamination procedures, but that won't be done for awhile yet.

It's not much, I know, but that's about it.

Tim Morken, B.A., EMT(MSA), HTL(ASCP)
Infectious Disease Pathology
Centers for Disease Control
MS-G32
1600 Clifton Rd.
Atlanta, GA 30333
USA

email: tim9@cdc.gov 
       timcdc@hotmail.com 

Phone: (404) 639-3964
FAX:  (404)639-3043



- ----Original Message Follows----
From: Susan McFarland <Susan.McFarland@lhsc.on.ca>
To: HistoNet@Pathology.swmed.edu 
Subject: cryostat decontamination
Date: Fri, 03 Mar 2000 14:01:16 -0500

Hello there,

I just did a search through the archives for procedures for disinfection of 
cryostats and could find only questions, not answers. Does anyone have a 
protocol, with references for adequately disinfecting their cryostats they 
would be willing to share?  If it makes any difference with have both old 
Ames cryostats and newer Leica 1720s.

I am in the process of trying to standardize protocols between two merged 
hospitals and find that neither protocol seems sufficient. I have nothing to 
make a reference to in either case.

Thanks very much,

Sue McFarland,
London Health Sciences Centre
London, Canada









______________________________________________________
Get Your Private, Free Email at http://www.hotmail.com 



----------------------------------------------------------------------

Date: 4 Mar 2000 10:15:26 -0600
From: "Molinari, Betsy" <BMolinari@heart.thi.tmc.edu>
Subject: RE: LAB  FUMES

Dear Sheila,
Once a year we have our fume hoods checked and certified, for proper air
exchange.  This is to meet GLP standards and for our own peace of mind.
We are affiliated with a hospital  so the Maintanence Dept. takes care
of it.

> ----------
> From:
> sheila.cleveland@orst.edu[SMTP:sheila.cleveland@orst.edu] 
> Sent: 	Friday, March 03, 2000 7:34 AM
> To: 	HistoNet Server
> Subject: 	LAB  FUMES
> 
> Hi,
> Is one sure they know if there lab is safe?  Sure hoods are 
> checked but what is safe??  I had a lab partner that became 
> chemicial sencitive and had to quiet her job.  If you can smell it, it
> 
> must be bad, xylene, hemo de, etoh's, if they don't smell thats 
> even worse.  I try and work with a window open all year long, just to 
> give myself peace of mind. 
> sheila.cleveland@orst.edu 
> 


----------------------------------------------------------------------

Date: 4 Mar 2000 11:00:21 -0600
From: Jazz1522@aol.com 
Subject: Job openings

There are a couple of openings for histotechs at US Labs in Irvine and Los 
Angeles, California.  If interested, fax resume to 949-450-0146 or call 949 
450-0145 ext. 114,  for Human Resources.


----------------------------------------------------------------------

Date: 4 Mar 2000 13:00:11 -0600
From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
Subject: using 100% as decontaminant in cryostat

I think the use of alcohol was discussed eons ago, as a decontaminating agent
in cryostat.  The question arose, 70% vs 95 or absolute alcohol, and 70% was
preferred for specific reasons, had something to do with suface tensions, etc.
hopefully someone will come forth with this again. 

Afer observing people inside of biohoods, cleaning down an area (wiping),
they use 70% ethanol to reduce contamination by bacteria and some are working
with adenoviruses.  I figured if they get no contamination of their cell
cultures, etc, with this type of cleaning, it would work in the cryostat as
well. 

To decontaminate, we defrost, wipe down with 70% several times on absorbant
towels, go to 100% to remove the residual water (cryostat is supposed to be
dry
before putting it back together)  There are some wipe cloths available now,
from Current Technology that look promising, if people want to do a final
wipedown after the 70% or even before that (when cryostat is warmed from
defrosting).  There are other sources of wipe cloths, but I was always afraid
to put too much gunk in the cryostat, didn't want corrosion.

We do wipe down areas in cryostat when it is running with 70% also, not a lot,
but enough to attract all the trimmings and after every use, if necessary.=20

I think this in one of the tough areas to deal with, cryostat innards and
protecting workers, and using formalin fumes just isn't acceptable.

Out of curiosity, does everyone cryosection with gloves on these days in the
clinical setting?

Gayle Callis




----------------------------------------------------------------------

Date: 4 Mar 2000 16:16:08 -0600
From: amos brooks <atbrooks@snet.net>
Subject: Re: cyclin D1

Hi Vinnie,
    We use Cyclin D1 frequently, on bone marrow biopsies. We use the DAKO
antibody (M7155 ?) diluted to 1:50.
The antigen is best retrieved using a high pH antigen retrieval. (also from
DAKO, I forget the pH) We use LSAB2
for a detection kit. Don't even bother with Envision+, it doesn't work. The
incubation times following
peroxidase blocking are 20 min primary, 10 min Link2, 10 min Label2 then the
chromogen you want. LSAB+ also
works but we phased this one out after a bad lot problem.
    The controls we use generally tend to be GI. We check each test we do to
determine if it can be used as a
control. If it is positive with out ridiculous amounts of background then we
cut extra slides to hold us over
until the next positive patient rolls along. If you are running low on
controls, and a doctor asks for lymphoma
markers on a GI case and not Cyclin D1 run one anyway just to check, that is a
good place to find them (Let the
doctor see it of course, never file a slide a doctor hasn't seen!)
    Just as a point of reference, I saw an incredible Cyclin D1 from a
representative of SIGNET the other day.
(made me drool all over the microscope what a mess :-) I think theirs blew
ours out of the water. There was
hardly any background staining, which is virtually impossible with this quirky
antibody. Unfortunately I dont
know what their detection method was. :-(
good luck
Amos Brooks

Vinnie Della Speranza wrote:

> I would appreciate hearing from anyone who has this marker working reliably
to learn
> 1. who's antibody you use
> 2. your antigen recovery method
> 3.  any caveats or tricks you might offer to provide better reliability
> 4. what you are using for a control
>
> we have had some cases stain very well, others that the pathologist insists
should be positive that weren't.
>
> Vinnie Della Speranza
> Manager for Anatomic Pathology Services
> Medical University of South Carolina
> 165 Ashley Avenue
> Suite 309
> Charleston, SC  29425
> ph:  (843) 792-6353
> fax: (843) 792-8974
> email: Dellav@musc.edu 




----------------------------------------------------------------------

Date: 4 Mar 2000 19:45:54 -0600
From: JIGPEP@aol.com 
Subject: Re: tissue cassettes and breast processing

Thanks to all who responded to my questions concerning the above topics. It 
was greatly appreciated and I will be using some of the info. Carol, 
concerning the yellow baskets from the old Technicon that you have, I will be 
glad to try them. I sent you an e-mail awhile ago, but perhaps it didn't go 
through. We were having some trouble with aol then. You can respond thru 
histonet if you are still interested in releasing those items.       Thanks a 
million!!!       Jeanne Godine
                                                          Pathology 
Consultants
                                                          Lynchburg VA


----------------------------------------------------------------------

Date: 4 Mar 2000 20:45:30 -0600
From: Victoria Baker <vbaker60@yahoo.com>
Subject: Re: using 100% as decontaminant in cryostat

Hi Gayle

Where I worked previously they were rigid with this
procedure.  Not using gloves could get a tech written
up for not following procedure. Clinical settings
leave the tech more unprotected from pathogens then
those in research.  We have an idea of what we have to
work with and the hazards it may impose.  Between TB
resurfacing,AIDS and other "goodies", universal
precaution is the rule.

Vikki Baker
American Health Foundation
Valhalla, New York


- --- Gayle Callis <uvsgc@msu.oscs.montana.edu> wrote:
> I think the use of alcohol was discussed eons ago,
> as a decontaminating agent
> in cryostat.  The question arose, 70% vs 95 or
> absolute alcohol, and 70% was
> preferred for specific reasons, had something to do
> with suface tensions, etc.
> hopefully someone will come forth with this again. 
> 
> Afer observing people inside of biohoods, cleaning
> down an area (wiping),
> they use 70% ethanol to reduce contamination by
> bacteria and some are working
> with adenoviruses.  I figured if they get no
> contamination of their cell
> cultures, etc, with this type of cleaning, it would
> work in the cryostat as
> well. 
> 
> To decontaminate, we defrost, wipe down with 70%
> several times on absorbant
> towels, go to 100% to remove the residual water
> (cryostat is supposed to be dry
> before putting it back together)  There are some
> wipe cloths available now,
> from Current Technology that look promising, if
> people want to do a final
> wipedown after the 70% or even before that (when
> cryostat is warmed from
> defrosting).  There are other sources of wipe
> cloths, but I was always afraid
> to put too much gunk in the cryostat, didn't want
> corrosion.
> 
> We do wipe down areas in cryostat when it is running
> with 70% also, not a lot,
> but enough to attract all the trimmings and after
> every use, if necessary. 
> 
> I think this in one of the tough areas to deal with,
> cryostat innards and
> protecting workers, and using formalin fumes just
> isn't acceptable.
> 
> Out of curiosity, does everyone cryosection with
> gloves on these days in the
> clinical setting?
> 
> Gayle Callis
> 
> 
> 
> 
__________________________________________________
Do You Yahoo!?
Talk to your friends online with Yahoo! Messenger.
http://im.yahoo.com 


----------------------------------------------------------------------

Date: 4 Mar 2000 22:15:28 -0600
From: wbtyler@webtv.net (WB Tyler)
Subject: Re: Azure B counterstain for melanocytic immunohistochemistry

Someone requested information on this.
This is the reference and the abstract.
J Cutan Pathol 1991 Dec;18(6):436-9 
Immunoperoxidase technique modified by counterstain with azure B as a
diagnostic aid in evaluating heavily pigmented melanocytic neoplasms.
Kamino H, Tam ST
Department of Dermatology, New York University Medical Center. 
Heavily-pigmented melanocytic neoplasms are difficult to evaluate on
routine hematoxylin and eosin stained slides because pigmented
melanocytes are difficult to distinguish from the numerous melanophages
that are usually seen in the background of these lesions.
Immunoperoxidase staining for S100 protein or HMB-45 antibody using
diaminobenzidine (DAB) as chromogen, which forms a brown product, does
not adequately distinguish melanocytes from melanophages. We modified
this technique by replacing hematoxylin as the counterstain with azure
B, which stains melanin green-blue. Thus, positive melanocytes appear
brown while melanin granules in their cytoplasm are green-blue. However,
negative melanophages only stain green-blue. This technique is useful in
evaluating heavily pigmented melanocytic lesions such as malignant
melanomas, melanosis of regressing malignant melanoma, residual
malignant melanoma in areas of granulation tissue with melanophages,
blue nevi, pigmented spindle cell variant of Spitz's nevi and combined
nevi. 
PMID: 1723081, UI: 92129698 

William B. Tyler MD
Department of Pathology
Geisinger Medical Center
Danville, PA



----------------------------------------------------------------------

Date: 4 Mar 2000 22:36:55 -0600
From: wbtyler@webtv.net (WB Tyler)
Subject: Re: Spleen microanatomy

Another reference for detailed spleen  histology is:
  
Histology for Pathologists  
by Stephen S., Md. Sternberg (Editor)  
 
Hardcover - 1248 pages 2nd edition (January 1998) 
Lippincott Williams & Wilkins Publishers; ISBN: 0397517181

This is available from Amazon.  $199

William B. Tyler MD
Dept of Pathology
Geisinger Medical Center
Danville, PA 



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