[Histonet] Miller's Elastin Stain

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Hi there,

Does anyone use commercial Miller's Elastic Stain other than VWR Ref 35115S one 
 ?
If so I would be grateful if you could let me know which one you use
Thanks in advance 

Malika



Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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[Histonet] H1B Visa Sponsorship

[Malika Benatti]

I wonder if anyone can help or perhaps knows anyone who could. I am a UK 
trained and registered medical technologist and have specialised in Histology, 
with 16 years laboratory experience. I am fully registered as a Specialist 
Biomedical Scientist with the Health Professional Council (Similar to the ASCP)

I am looking for a H1B Visa Sponsorship opportunity inTennessee/Knoxville or 
surrounding areas. I wonder if anyone can help or would be interested in having 
a informal discussion. I will be in Knoxville the last week September would be 
more then happy to travel and meet for an informal interview. 

Malika
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[Histonet] H1B Visa Sponsorship

[Malika Benatti]

I wonder if anyone can help or perhaps knows anyone who could. I am a UK 
trained and registered medical technologist and have specialised in Histology, 
with 8 years of experience. I am fully registered with theHealth Professional 
Council. 

I am looking for a H1B Visa Sponsorship opportunity inTennessee/Knoxville or 
surrounding areas. I wonder if anyone can help or would be interested in having 
a informal discussion. I will be inKnoxville between the 20th of September 
until the 2nd of October and would be more then happy to travel and meet for an 
informal interview. 

Malika
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[Histonet] Olig2 (ab33427) on the bond max

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

I was wondering if anyone on the histonet list is using abCam Olig2 (ab33427) 
on the bond max.
I have been trying various titration and enzyme treatment on surgical brain 
tissue with not much success al we seams to achieve is nuclear staining and 
heavy background staining. 

Any suggestions would be appreciated! 

Thanks in advance 

Malika


 


Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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[Histonet] Lyve - 1 on Bond Max

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Hi there,

I was wondering if anyone on the histonet list use anti-mLYVE-1 antibody 
(purified in rat Monoclonal IgG 2A from R&D System) on the Bond Max.
We have been trying out various titrations and pre-treatment without much 
success.
All we seems to achieve is background staining with no real specificity at 
1:100 H1(10) ; 1:100 H2(20) ;1:100 E2(10): 1:50 E1(30) ;1:50 H1(30); 1:50 H2(10)

Any suggestion ?

Regards

Malika










Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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[Histonet] RE: Coverslips

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Hi there,

We use the CellPath CoverSlips 22x50mm No 1.5 they are great for hand
mounting but when used with the Leica CV5030 they are a real pain.
most of the CoversSlips get rejected.

If anyone use the same equipment, let me know how you are getting one
.

Cheers,

Malika





--

Message: 7
Date: Mon, 28 Jun 2010 15:06:08 -0400
From: "Weems, Joyce" 
Subject: [Histonet] RE: Coverslips
To: "histonet@lists.utsouthwestern.edu"

Message-ID:
<92ad9b20a6c38c4587a9febe3a30e164015dfd5...@chexcms10.one.ads.che.org>
Content-Type: text/plain; charset="us-ascii"

I forgot to say that the packaging has changed to their Histo Hippo
theme, but they assure me they are the same product. j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Weems, Joyce
Sent: Monday, June 28, 2010 14:29
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Coverslips

For years we have used coverslips from Mercedes - the ones in a plain
white box labeled in German. This year we have begun to have problems
with them being dirty and having bits of glass on them that cause air
bubbles when used on the automatic coverslipper. Is anyone else having
this problem? They have worked with us to try to resolve it, and have
replaced several boxes, but the replacements are doing the same thing.
Thanks for your input, j


Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
678-843-7376 - Phone
678-843-7831 - Fax


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Message: 8
Date: Mon, 28 Jun 2010 12:21:33 -0700
From: "Kathleen Boozer" 
Subject: Re: [Histonet] Coverslips
To: "histonet@lists.utsouthwestern.edu"
,"Joyce Weems"

Message-ID: <4c2893cc.4aa8.00c...@ah.org>
Content-Type: text/plain; charset=US-ASCII

I am having the same issues and we purchased the expensive "Platinum"
to avoid that problem.  They just sent me replacements and I haven't
tested them all yet.  

Kathy Boozer, HT (ASCP), IHCQ
Adventist Medical Center
10123 SE Market St.
Portland, OR  97216
booze...@ah.org 

 


Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


>>> "Weems, Joyce"  6/28/2010 11:28 AM >>>
For years we have used coverslips from Mercedes - the ones in a plain
white box labeled in German. This year we have begun to have problems
with them being dirty and having bits of glass on them that cause air
bubbles when used on the automatic coverslipper. Is anyone else having
this problem? They have worked with us to try to resolve it, and have
replaced several boxes, but the replacements are doing the same thing.
Thanks for your input, j


Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
678-843-7376 - Phone
678-843-7831 - Fax


Confidentiality Notice:
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not the intended recipient, please delete this message, and 
reply to the sender regarding the error in a separate email. 
 
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--

Message: 9
Date: Mon, 28 Jun 2010 15:36:22 -0400
From: "Sherwood, Margaret " 
Subject: RE: [Histonet] RE

[Histonet] H1B Visa Sponsorship

[Malika Benatti]

To all Histonet list members in Knoxville Area 

I am Specialist Biomedical Scientist fully trained in the United Kingdom with a 
BSc Hons in Biomedical Science and a Post Graduate Certificate in Cellular 
Pathology from the University of Westminster in London United Kingdom and 
currently looking for a H1B Sponsor to work as a histotechnologist, as I 
currently looking to relocate in the United States in Knoxville Area.

I am fully registered with the Health Professional council in the UK (Similar 
to the US, ASCP Certification), with 8 years Post HPC Registration. I am fully 
trained in all the aspect of routine histology technical work and I have a very 
strong background in pediatric pathology, and 9 years experience in general 
pathology.  including cut-up, special stains, immunocytochemistry, urgent intra 
operative frozen sections, neuropathology, muscle biopsy, and electron 
microscopy tissue processing. 


Malika

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[Histonet] Glut1 antibody

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **


We use anti-glut-1 (rabbit polyclonal) Purchased from Millipore

http://www.millipore.com/catalogue/item/07-1401 

Hope this helps,

Malika



--

Message: 7
Date: Tue, 22 Jun 2010 16:10:12 -0400
From: "Martha Ward" 
Subject: [Histonet] Glut1 antibody
To: 
Message-ID:
<61135f0455d33347b5aae209b903a30433deb...@exchvs2.medctr.ad.wfubmc.edu>

Content-Type: text/plain;   charset="us-ascii"

I have been using anti-human Glut-1 from Dako for several years but when
I went to order it I was told it was discontinued.  Does anyone have a
suggestion of where I can locate this antibody?  Thanks in advance for
your help!
 
Martha Ward, MT (ASCP) QIHC
Assistant Manager, Molecular Diagnostics Lab
Wake Forest University Baptist Medical Center
336-716-2104
 
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End of Histonet Digest, Vol 79, Issue 28
********

Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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[Histonet] Gordon & Sweet Reticulin Stain problem on BMT Sections

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Dear all,

Lately we have been experiencing problem with our Retic Stains, who keep 
lifting off slides after the gold chloride step.

We have changed all solutions one by one, trying to see which one was causing 
problem without success.
BMT Bx are cut at 3 micron on X-tra adhesive slides.
BMT BX are decal in 10 % Formic Formaldehyde.

Here is the protocol that we are currently using.

1.   Sections to water.
2.   Treat with 0.25% acidified Potassium permanganate. --- 5 mins
3.   Bleach in oxalic acid.
4.   Rinse in distilled water.
5.   Treat with 5% iron alum.   --- 15 mins 
   
6.   Rinse briefly in distilled water.
7.   Impregnate with filtered ammoniacal silver solution  ---1 min
NB: Time in ammoniacal silver may need to be increased up to 5 min for bone 
marrow trephines
8.   Rinse well in distilled water.
9.   Reduce in 10% formalin (in tap water). --- 1min
10.  Rinse in distilled water.
11.  Tone in 0.2% Gold Chloride  ---1 min
12.  Rinse in distilled water.
13.  Fix in 5%  sodium thiosulphate (hypo)  --- 30 sec
14.  Wash in water.
15.  Counterstain with neutral red.
16.  Dehydrate, clear and mount. 


Any suggestions would be  appreciated.

Malika


Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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[Histonet] Alpha Dystroglycan Immuno

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **


Having issue with Millipore Alpha Dystroglycan antibody.

Millipore recommend 1:100 dilution so we  tried various titrations ranging from 
1:100, 3:200 to 9:200, normal and abnormal muscle tissue with very poor 
results. After staining up to  9:200 dilution immuno come out patchy not strong 
enough

Any one on the histonet list use this antibody, if so at what dilution.

any input would be really appreciated

Malika



Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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Re: [Histonet] Bone marrow trephine decalcification

[Malika Benatti]

Goodings & Stewarts also known as Formic/Formaldehyde fluid (10%  
formic acid, 5% formalin) for 6 hours  should allow decal of fix BMT  
trephine Bx.





" ... Smile it confuses people ..."

On 21 Apr 2010, at 21:14, "Marinez"  wrote:

I'm in a bit of trouble. I work in the south of Brazil (Porto  
Alegre) for some years and now in my hospital a lot of bone marrow  
biopsies are being performed (leukemia and lymphomas). With  
decalcification with strong acids (nitric) I'm getting very poor  
results with the immunohistochemistry. I'm trying to use EDTA but we  
really are not getting speed or good results (we  controled de pH  
but still precipitates). Could some one be kind enough to send me  
some formula (simple one please) that will work in reasonable time  
(24 or 48 hours) and perform well with immunohistochemistry? I would  
deeply grateful.




M. Barra
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[Histonet] Chloroform Fume Monitoring

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Hi Histoneter,

I was just wondering what kind Health and Safety of procedures do people use to 
monitor Chloroform Fume ?

I was carrying an H&S audit for occupational exposure  Xylene / Glutaraldehyde 
/ Formaldehyde/ Chloroforms fumes using Surgipath Exposures monitoring badge on 
a 3 months interval. when I realised that we do not have any Chloroform Badges 
available, and will not be able to get some for a little while. 

Does anyone use these EMT's Chloroform Monitoring Kit available to purchase 
from this site http://www.emt-online.com/ProductPages/Chloroform.htm 
Or is there any better alternative ?

Malika




Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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Re: [Histonet] Dark background with Gomori's reticulin impregnation

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Andre,

By looking at your method it does not look right from the start.
We use the Gordon and Sweet Method and it work wonder for demonstration
of reticulin fibers
Here it is Hope it helps

Malika

1.   Sections to water.
2.   Treat with 0.25% acidified Potassium permanganate. --- 5
mins
3.   Bleach in 1% oxalic acid.
4.   Rinse in distilled water.
5.   Treat with 5% iron alum.   --- 15 mins
6.   Rinse briefly in distilled water.
7.   Impregnate with ammoniacal silver solution. Filter Solution before
use  ---1 min
(Note:  Time in ammoniacal silver may need to be increased for bone
marrow trephines) 
8.   Rinse well in distilled water.
9.   Reduce in 10% formalin (in tap water). --- 1min
10.  Rinse in distilled water.
11.  Tone in 0.2% Gold Chloride ---1 min
12.  Rinse in distilled water.
13.  Fix in 5%  sodium thiosulphate (hypo)  --- 30 sec
14.  Wash in water.
15.  Counterstain with neutral red.
16.  Dehydrate, clear and mount.

Results
Reticulin   - Black
Collagen- Grey
Nuclei  - Red




> From: André Kougioumoutzakis 
> Date: 18 April 2010 23:08:35 GMT+01:00
> To: 
> Subject: [Histonet] Dark background with Gomori's reticulin  
> impregnation
>

>
> Hi everyone,
>
>
>
> I've been performing Gomori's reticulin impregnation and never got  
> the results I see in textbooks. The reticulin fibers stain fine, but 

> I get a dark purple/gray background instead of a light pink  
> background. Does anyone know why ?
>
>
>
>
>
> Here is the technique I use :
>
>
>
> 1) Deparaffinize and rehydrate
>
> 2) 0,5% potassium permanganate   1 min
>
> 3) Running tapwater 3 min
>
> 4) Distilled water
>
> 5) 5% Oxalic acid 2 min
>
> 6) Running tapwater 3 min
>
> 7) Distilled water
>
> 8) 2% ammonium iron sulfate  1 min
>
> 9) Running tapwater 3 min
>
> 10) Distilled water
>
> 11) Ammoniacal silver1 min
>
> 12) Distilled water15 sec
>
> 13) 20% Formaldehyde  3 min
>
> 14) Running tapwater3 min
>
> 15) Distilled water
>
> 16) 0,2% Gold chloride   5 min
>
> 17) Distilled water
>
> 18) 3% potassium metabisuphite  5 min
>
> 19) Distilled water
>
> 20) Sodium thiosulfate   1 min
>
> 21) Running tapwater3 min
>
> 22) Distilled water
>
> 23) 0,1% Nuclear fast red 15 min
>
> 24) Distilled water
>
> 25) Dehydrate, clear and mount
>
>
>
> Thanks for your help!
>
> Andre
>
>
>
> _
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Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


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Re: [Histonet] Communication when pathologist are not in same area as lab

[Malika Benatti]

Oops it's looking like I hit the send key too fast ;)

Email request can be problematic as very often request are received after
hours.

But if you give me the choice between email request and face to face
communication, like you mentioned Jan it definitely depend on the
individuals involved.

M



On Wed, Apr 14, 2010 at 8:47 PM, Mahoney,Janice A <
janice.maho...@alegent.org> wrote:

> With the right LIS and standard work you may never have to or get to,
> depending on your perspective, speak with a Pathologist again.
> Jan
> Omaha
>
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu [mailto:
> histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Malika Benatti
> Sent: Wednesday, April 14, 2010 2:38 PM
> To: Scott, Allison D
> Cc: histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] Communication when pathologist are not in same area
> as lab
>
> Well our 5 pathologists, advance practitioner and SPR are just across the
> way from us, until last July all request were made personally by
> pathologist
> to staff and if there were any query with regard to the request made it was
> sorted then and then and it was pretty straight forward.
>
> But the good all days of face to face communication changed when we started
> to get requests via email, although a good idea, in principle, requests via
> email are not without pitfalls and therefore face to face communication
> between lab staff and pathologist is still necessary.
>
>
> On Wed, Apr 14, 2010 at 4:09 PM, Scott, Allison D <
> allison_sc...@hchd.tmc.edu> wrote:
>
> > Hello to all in histoland.  What are you doing to minimize the
> > communication breakdown, when the pathologist are not housed in the lab.
> > We will be moving to a new lab and the pathologist will not be in the
> > new bldg with us.  They will be in the old building next door. Are you
> > relying upon email and phone interactions.   My directors concern is how
> > will this effect the flow of information, especially when they are used
> > to face to face interactions.
> >
> > Allison Scott HT(ASCP)
> > LBJ Hospital
> > 5656 Kelley
> > Houston, Texas 77026
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>
>
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Re: [Histonet] Communication when pathologist are not in same area as lab

[Malika Benatti]

Well our 5 pathologists, advance practitioner and SPR are just across the
way from us, until last July all request were made personally by pathologist
to staff and if there were any query with regard to the request made it was
sorted then and then and it was pretty straight forward.

But the good all days of face to face communication changed when we started
to get requests via email, although a good idea, in principle, requests via
email are not without pitfalls and therefore face to face communication
between lab staff and pathologist is still necessary.



 say email request can be problematic as very often request.

On Wed, Apr 14, 2010 at 4:09 PM, Scott, Allison D <
allison_sc...@hchd.tmc.edu> wrote:

> Hello to all in histoland.  What are you doing to minimize the
> communication breakdown, when the pathologist are not housed in the lab.
> We will be moving to a new lab and the pathologist will not be in the
> new bldg with us.  They will be in the old building next door. Are you
> relying upon email and phone interactions.   My directors concern is how
> will this effect the flow of information, especially when they are used
> to face to face interactions.
>
> Allison Scott HT(ASCP)
> LBJ Hospital
> 5656 Kelley
> Houston, Texas 77026
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Re: FW: [Histonet] studying for ASCP certification exam

[Malika Benatti]

Theory and Practice of Histological Techniques by John D. Bancroft and
Marilyn Gamble Dr. you can probably get hold of a copy on Amazon for 1/2
price, even at full price it definitely worth every cents.




On Wed, Apr 14, 2010 at 6:32 PM, Kim Merriam  wrote:

> The NSH sells a set of study guides on histology (maybe 10 of them, on
> fixation, various special stain categories and the like), I think you will
> find them quite helpful to see how much you actually remember after reading
> the textbooks.
>
> Kim
>  Kim Merriam, MA, HT(ASCP)QIHC
> Cambridge, MA
>
>
>
>
> 
> From: Pat Laurie 
> To: histonet@lists.utsouthwestern.edu
> Sent: Wed, April 14, 2010 12:02:21 PM
> Subject: Re: FW: [Histonet] studying for ASCP certification exam
>
> I definetly agree with the previous suggestions.  Those 3 books are in the
> top 5 of my collections.  Another book that might help you because of the
> illustrations (special stain color pictures on the test!) is:
>
> Theory and Practice of Histological Techniques
> 5th edition edited by Bancroft and Gamble
>
> Unfortunately it is rather pricey.
>
>
> Another good standby because so many books seem to cite it (classic):
>
> AFIP Laboratory Methods in Histotechnology
> latest edition is Prophet et.al. ISBN: 1-881041-00-X
>
> Good luck with your test!
> On Wed, Apr 14, 2010 at 6:58 AM, Ian Montgomery <
> ian.montgom...@bio.gla.ac.uk> wrote:
>
> > Jennifer,
> >
> > HISTOLOGICAL AND HISTOCHEMICAL METHODS. 4th Ed. Kiernan, J.A.
> >
> >In my opinion this is the best histotechnique text currently
> > available. Amazon UK has it in stock for £34.99 and for a text of this
> > quality it's a bargain. If you are studying for an exam this text gives
> > everything you'll need, but more importantly, it is written in a style
> that
> > is easily understood. Better still, when you have finished studying, the
> > text can sit on a shelf in your lab and become your standard lab manual.
> No
> > tie in with the author or publisher but I have 2 copies, one in my office
> > and the other in the lab.
> > Ian.
> >
> > Dr. Ian Montgomery,
> > Histotechnology,
> > I.B.L.S. Support Unit,
> > Thomson Building,
> > University of Glasgow,
> > Glasgow,
> > G12 8QQ.
> > -Original Message-
> > From: histonet-boun...@lists.utsouthwestern.edu
> >  [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike
> > Pence
> > Sent: 14 April 2010 14:18
> > To: Jennifer Campbell; histonet@lists.utsouthwestern.edu
> > Subject: RE: [Histonet] studying for ASCP certification exam
> >
> > Also try:
> >
> > "Theory and Practice of Histotechnology" by Sheehan and Hrapchak.
> >
> > I have heard this is a VERY hard book to get a hold of these days and if
> > you can it will cost!
> >
> > -Original Message-
> > From: histonet-boun...@lists.utsouthwestern.edu
> > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer
> > Campbell
> > Sent: Tuesday, April 13, 2010 6:46 PM
> > To: histonet@lists.utsouthwestern.edu
> > Subject: [Histonet] studying for ASCP certification exam
> >
> >
> >  I am in the process of studying for my HT certification exam and was
> > wondering if anyone had any recommendations on text books or study
> > guides they found to be helpful.  I am currently studying
> > "Histotechnology: A Self-Instructional Text", by Carson and just
> > recieved the "BOR study Guide for Histotechnology".  Are there any other
> > sources you would recommend?  I have taken a look at the suggested
> > reading list on the ASCP website but, there are quite a few books
> > listed.
> >
> > Thanks in advance,
> >
> > Jennifer Campbell ___
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
> >
> > ___
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> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
> > ___
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> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
>
>
>
> --
> Patrick Laurie HT(ASCP)QIHC
> CellNetix Pathology & Laboratories
> 1124 Columbia Street, Suite 200
> Seattle, WA 98104
> PH: 206-215-5949
> plau...@cellnetix.com
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>
>
>
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Re: [Histonet] Masson Trichome Staining: Can I use Fast Green?

[Malika Benatti]

** Proprietary **
** Reply Requested When Convenient **

Colin,

If you havin difficulty with MT try the Gomori One Step 

1.  Sections to water.
2.  Pretreat in 3% Potassium dichromate for 1hr at 60ºC 
3.  Stain nuclei with celestin blue/Mayer's haematoxylin. 5  min
4.  Wash in tap water.
5.  Differentiate and blue.
6.  Stain with chromotrope green mixture  --- 10 mins.
7.  Rinse in distilled water
8.  Blot and dehydrate from absolute alcohol ,clear and mount. 

Results
Muscle and fibrin   -  Red
RBC's   -  Red
Collagen-  Green

MASSON TRICHROME (ONE-STEP)
Ingredients:
Chromotrope 2R  0.6 g.
Fast Green FCF  0.3 g.
Phosphotungstic acid0.6 g.
Glacial acetic acid 1 ml.
Distilled water 100 ml.

METHOD:-
Dissolve the chromotrope 2R, the fast green and the phosphotungstic
acid in the water and add the glacial acetic acid.

Hope this help


Malika Benatti
 
Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH

 
Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875
 
ben...@gosh.nhs.uk


>>> John Kiernan  09/04/2010 17:08 >>>
"The slides have not been coming out well." is not very informative!
Your problems may have nothing to do with which dye you are using as the
collagen stain. There are plenty of other things that can make a
multi-step trichrome method give unexpected results. 
 
Fast green FCF (C.I. 42053) is usually considered superior to light
green SF (C.I. 42095) on account of being less prone to fading. Be sure
you have the correct dye, and that it is from a batch certified by the
Biological Stain Commission (B.S.C. - see the label on the bottle of dye
powder, which will also tell you the dye content). If you buy a
ready-made solution, check with the supplier. 
 
The minimum acceptable dye content for certification of fast green FCF
(85%) is higher than the minimum for light green SF (65%). A 2% solution
of the former might therefore have greater tinctorial power than a 2%
solution of the latter. The B.S.C. tests both dyes as substitutes for
aniline blue in Gomori's one-step trichrome method; the concentration of
blue or green dye in the mixture is 0.3%, irrespective of the dye
content of the powder.  Gomori's is a more stringent test for dyes than
Masson's trichrome method because there is no visual/manual control of
the staining. A dye that works with Gomori's method should work well
with Masson's. 
 
The Masson variant that you have been trying (from Bryan Llewellyn's
excellent web site, 
http://stainsfile.info/StainsFile/stain/conektv/masson.htm) is well
documented. Follow up some references from Bryan's citation of the
Bancroft & Stevens book, and learn the reasons for all the 11 steps of
the modified Masson technique. The current edition (6th, 2008) of
"Theory and Practice" is edited by Bancroft & Gamble: ISBN
9780443102790.
 
My textbook (ISBN 97819048422) covers trichrome methods in Chapter 8,
with plenty of references to scientific/scholarly literature, much of
which is now available on the internet, especially if you can go through
a university or public library's web interface. There is a well
ilustrated chapter on troubleshooting trichromes, by Vinnie Della
Speranza, in R.W.Brown's (2009) "Common Problems" book, ISBN
9780930304959 (pp. 95-101).
 
A rinse in tap water at any stage after after washing out the
iron-haematoxylin nuclear stain can weaken either the red or the
blue/green component of any trichrome method. So can "graded alcohols"
for the final dehydration of well stained slides. 
 
John Kiernan
Anatomy, UWO
London, Canada
= = =
- Original Message -
From: cscam...@uci.edu 
Date: Thursday, April 8, 2010 18:42
Subject: [Histonet] Masson Trichome Staining: Can I use Fast Green?
To: HistoNet 

> Hi Histonet,
> 
> I am currently using the protocol from Stainsfile for staining heart
> tissue: http://stainsfile.info/StainsFile/stain/conektv/masson.htm 
> 
> My lab had Fast Green readily available so I substituted it for
Light
> Green in Solution C. The slides have not been coming out well. I 
> failed to
> account for the difference in timing between FG and LG - running 
> the slide
> in FG for the full 10 minutes. Has anyone used FG in Masson's 
> Trichrome?If so, how long did it take to get the desired colors 
> in the solution? Or,
> is Fast Green a poor substitute for Light Green?
> 
> Thanks for your advice!
> -Colin
> 
> 
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Re: [Histonet] 72644.18148...@web111105.mail.gq1.yahoo.com

[Malika Benatti]

Bryan,

Thanks for your details explanation on what look to be a very different
system.
Though something I am confused with an maybe you or someone else can clarify
this for me.

In the UK to be allow to practice as a Biomedical Scientist in any
laboratory discipline, you need of have a Accredited Hons Degree, and
undergo your HPC registration, Specialist status is only acquire after
passing the part 2 of the HPC registration (this is pretty new ).

Most of fully qualify Specialist Biomedical Scientist have also Post
Graduate qualifications.

Now I am reading right that not all the histotechnologist in the US are ASCP
registered ?
Is the ASCP registration not a Mendatory requirement to practice as a
histotechnologist ?
Does the registration rules vary from states to states?

Cheers

Malika





On Mon, Apr 5, 2010 at 8:37 PM, Bryan Llewellyn  wrote:

> Comparing the situations in different countries can be very confusing.  I
> traied in the UK (many years ago) and have lived in Canada for a long time,
> but I do have some (limited) information about the US system.
>
> First off, Medical Laboratory Technology in the UK and Canada includes
> histotechnology as one of the integral subject areas, but the US does not.
> Histotechnology is a standalone subject there, by and large.
>
> The ASCP is different from the Health Professional Council (used to be
> Council for professions supplementary to medicine when I lived there).  That
> is a licensing body, and its function is carried out by some state agencies
> (in the US) and some provincial agencies (in Canada).  However, not all
> states and provinces require licensing to work as a medical laboratory
> technologist/histotechnologist.  The ASCP used to run the commonest US
> qualifying exams (still do ??, I am not sure) and kept a registry of
> qualified technologists, although there are others systems.  In Canada it is
> done by the CSMLS (Canadian Society for Medical Laboratory Sciences).  The
> equivalent organisation in the UK is the IBMS (Institute of Biomedical
> Sciences).
>
> In Canada it is possible to take specialty training in a subject area at
> both initial level and post initial level.  So you can be an RT (Registered
> Technologist) in medical laboratory technology generally, or an RT in
> cytology or electron microscopy as examples.  All RTs can take advanced
> examinations as general or specialist technologists, depending on their
> initial RT status.  There used to be a third level (Fellowship in the CSMLS)
> but it was abandoned because so few technologists took it.  That was about
> the same level as the UK three part exam.  An applicable BSc is now required
> in Canada to advance post RT.
>
> As to 16 year olds in labs.  I started work in the UK in Hackney six days
> before my 17th birthday in 1960.  A month later I was doing haemoglobins by
> finger stick with Hagedorn needles, ESRs and going around the wards.  A year
> later I was well versed in clinical chemistry (urea, glucose, bilirubin,
> alkaline phosphatases etc - all done manually with what passed for micro
> methods in those days.  Students like me did about 80% of the work in those
> days because the profession was expanding so fast due to the introduction of
> the public health care system in Britain.  Things change, and that just
> would not be allowed today.  I suspect that 16 year olds doing grossing is
> very unusal and in the US would likely be viewed as an invitation for the
> pathologists to be sued.  Remember, it was April 1st!
>
> In the US the CAP (College of American Pathologists) is involved in an
> accreditation system with other agencies.  In Canada the CAP (Canadian
> Association of Pathologists) is a player in some accrediatation systems, but
> in Canada health care is legally a provincial responsibility, so
> accreditation is done by provincial agencies for each province.  That is
> also the reason licensing varies from province to province here.  Our
> country wide qualifying system by the CSMLS is a fortunate anomoly that
> nobody wants to change because it works so well for us.
>
> I hope this explains a little.
>
> Bryan Llewellyn
>
> - Original Message - From: "Malika Benatti" <
> malbena...@googlemail.com>
> To: "Mark Tarango" 
> Cc: ; "Andrew Burgeson" <
> nap...@siscom.net>
> Sent: Monday, April 05, 2010 11:03 AM
> Subject: Re: [Histonet] 72644.18148...@web05.mail.gq1.yahoo.com
>
>
>  I am very confuse reading every email reply to this tread also I would be
>> really grateful if someone could enlighten with regard to what is the
>> comment practice in the US.
>>
>> Having been trained as a histotechnologist although we are call Specialist
>> Biomedical Scientist in the UK, we ca

Re: [Histonet] 72644.18148...@web111105.mail.gq1.yahoo.com

[Malika Benatti]

I am very confuse reading every email reply to this tread also I would be
really grateful if someone could enlighten with regard to what is the
comment practice in the US.

Having been trained as a histotechnologist although we are call Specialist
Biomedical Scientist in the UK, we cannot practice unless we are fully
registered with the Health Professional Council HPC, which I believe has the
same role as the ASCP. Every 2 years we may be audited a demonstrate that we
fully comply with HPC regulation and CPD or lose or registration. All
laboratories are accredited by the Clinical Pathology Accreditation CPA
under the international organization for standardization legislation (ISO
15189).

Laboratory accreditation happen every 2 years cycle for which the laboratory
has to comply with a set of standard.
During inspection accessor review everything with a fine tooth comb, and
score you some of the issues may just be minors but they will always get you
with a critical issue, which you will have a set amount of time to correct,
they will then return and verify that all non compliance and critical issues
have been address before giving you CPA accreditation status.

Having a 16 years old out of school with little experience in histology and
no formal training grossing specimen is never heard off, only Register
Biomedical Scientist are allowed to do small biopsies, Advance Practitioner,
Trainee Pathologist, will be involved in the grossing of lager specimens,
and tumour specimens.




On Mon, Apr 5, 2010 at 6:18 PM, Mark Tarango  wrote:

> When I was 16 years-old I was grossing in the lab.  We had
> very busy pathologists (busy reading slides) who thought it was okay to
> train their courier (me at the time) to gross.  I had already been there
> handing them bottles and closing cassette lids for several months.
>
> When we had our first CLIA inspection, they had me intial and sign
> paperwork
> saying that I had grossed so many cases of various specimen types under
> patholgist supervision and had been grossing so long.  The problem was that
> I was only a high school graduate at the time.  They then changed direction
> and told the inspectors that the pathologists did all the grossing.
>
> Just brings back memories.  Thought I'd share.
>
> Mark Tarango
> On Mon, Apr 5, 2010 at 10:03 AM, Andrew Burgeson 
> wrote:
>
> > The point is not about gender, as I stated before...
> >
> > It's about a person's health risks and lack of training
> > overlooked for the sake of labor. TYVM
> >
> >
> >
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> >
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Re: [Histonet] Histobath

[Malika Benatti]

Sorry for that Matt,

In this case the internal temperature of the cryostat chamber should be set
at -20 to -22 degree Celsius, not sure what would the actual temperature of
the histobath itself but our thermo scientific cryostat has the following
settings:

   - the cryobar is set at -18 Degree Celsius,
   - the Cryo chamber at - 22 degree Celsius.


Hope this help

Malika Benatti

Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London, WC1N 3JH
UK

Tel: (+44) 0207 4059200 Ext 5475
ben...@gosh.nhs.uk
- Show quoted text -

On Mon, Apr 5, 2010 at 1:51 PM, Matthew Mincer  wrote:

> Thanks, but the Histobath I was talking about is a freezing device for
> cryostats.
>
> Matt
>
> Malika Benatti wrote:
>
>> Hi Matt,
>>
>> Depending on the wax melting point which should be around 57 degree
>> Celsius, your water bath should be set at 50 degree Celsius.
>>
>> Hope this help.
>>
>> Malika Benatti BSc MIBMS
>> Specialist Biomedical Scientist
>> Great Ormond Street Children Hospital
>> London, WC1N 3JH
>> UK
>>
>> Tel: (+44) 0207 4059200 Ext 5475
>> ben...@gosh.nhs.uk <mailto:ben...@gosh.nhs.uk>
>>
>>
>>
>>
>>
>> On Mon, Apr 5, 2010 at 1:37 PM, Matthew Mincer > m...@techoneweb.com>> wrote:
>>
>>Does anyone know the "normal" operating temperature of the Thermo
>>Histobath? A customer has asked me to repair theirs and I want to
>>make sure I get the temp correct.
>>
>>Thanks
>>Matt
>>
>>-- Matthew Mincer
>>Tech One Biomedical Service
>>159 N Marion Street
>>PMB163
>>Oak Park, IL 60301
>>office  (708) 383-6040 X 10
>>cell(708) 822-3738
>>
>>
>>___
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>><mailto:Histonet@lists.utsouthwestern.edu>
>>
>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>>
>>
>> --
>> " Smile  it confuses people  "
>>
>
>
> --
> Matthew Mincer
> Tech One Biomedical Service
> 159 N Marion Street
> PMB163
> Oak Park, IL 60301
> office  (708) 383-6040 X 10
> cell(708) 822-3738
>
>


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Re: [Histonet] Histobath

[Malika Benatti]

Hi Matt,

Depending on the wax melting point which should be around 57 degree Celsius,
your water bath should be set at 50 degree Celsius.

Hope this help.

Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London, WC1N 3JH
UK

Tel: (+44) 0207 4059200 Ext 5475
ben...@gosh.nhs.uk




On Mon, Apr 5, 2010 at 1:37 PM, Matthew Mincer  wrote:

> Does anyone know the "normal" operating temperature of the Thermo
> Histobath? A customer has asked me to repair theirs and I want to make sure
> I get the temp correct.
>
> Thanks
> Matt
>
> --
> Matthew Mincer
> Tech One Biomedical Service
> 159 N Marion Street
> PMB163
> Oak Park, IL 60301
> office  (708) 383-6040 X 10
> cell(708) 822-3738
>
>
> ___
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> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



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Re: [Histonet] cassette labels erased by processor

[Malika Benatti]

Another option would be to invest in a cassette labeling machine, they  
are few a available in the market, we use the Surgipath Cassette  
Marker, they are pretty good as the ink fix with formalin and survive  
processing.
Though ribbon need to keep refrigirated when not in use(overnight and  
weekend) and printing heard need to be clean at least once a week, but  
apart from tha it is pretty straight forward to use,
also it can be program so it make printing a large number of cassette  
painless .


I have also used in tha past cassette marker that use thermal print  
ribbon so again no risk of lab number not surviving processing.So if  
you lab as some spare cash to spend  get few on trial in, az they are  
worth it.


Best wishes,

Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London

 " ... Smile it confuses people ..."

On 1 Apr 2010, at 00:24, Amos Brooks  wrote:


Indeed,
 One of the most EVIL products ever designed is the "Permanent  
Lab Marker" by VWR. Talk about false advertising! We have  
researchers drop off buckets of cassettes that they have labeled  
using these wonderful products. Pencil is the ONLY acceptable means  
of labeling cassettes. It is just not worth the risk to use anything  
else. (Except cassette labelers they are specifically designed for  
this and have a sales rep you can beat up if they fail.) Even pens  
specifically designed for cassettes have failed at times.


Just my $0.02,
Amos


Message: 23
Date: Wed, 31 Mar 2010 15:40:44 +0100
From: Malika Benatti 
Subject: Re: [Histonet] cassette labels erased by processor
To: Catherine Breen 
Cc: Histonet List 
Message-ID: 
Content-Type: text/plain;   charset=us-ascii;
format=flowed;  delsp=yes


Hi there,
I would suggest to use a pencil rather than a marker pen to label
cassettes when processing cassette with the Sakura VIP , my lab had a
number of the so called solvent proof pen on trial and have yet to
find one that survive processing.

Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London

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Re: [Histonet] GMS stain

[Malika Benatti]

Hi Margaret,

What problem are you experiencing ?
Are you over impregnated or under impregnated ?
At what thickness do you cut your Section ? Ideally 3 to 4 microns
How do you make up you Hexamine solution ? if you get contamination of
Hexamine solution make sure that glass ware is clean.

Try this.
Make 0.75 g of hexamine in 100 ml coplin jar / leave it dissolve in
distilled H2O at 60 oC while your slides are in chromic acid. When ready
slowly ad 18 drops (approximately 1ml) of 5% silver nitrate, (make sure
solution remain clear if cloudy, then glassware is contaminated and repeat
this steps with chromic acid clean glassware) then add 2 ml of borax. Sliver
solution should remain clear. add slides and check macroscopically slide
after 5 to 7 mins, then at 5 mins interval after that. Depending on tissue,
it should not take more that 15 mins to develop, then carry to next step.

Hope this help

Best wishes

Malika


Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Hospital
London, UK


On Wed, Mar 31, 2010 at 7:46 PM, Perry, Margaret  wrote:

> We sometimes have problems with the stain if we use positive slides.
> Margaret
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Re: [Histonet] cassette labels erased by processor

[Malika Benatti]

Hi there,

I would suggest to use a pencil rather than a marker pen to label  
cassettes when processing cassette with the Sakura VIP , my lab had a  
number of the so called solvent proof pen on trial and have yet to  
find one that survive processing.





Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London

 " ... Smile it confuses people ..."

On 31 Mar 2010, at 15:13, "Catherine Breen"  wrote:

I am looking for help solving a lab mystery.  The cassettes in our  
lab are labeled with an SP Securline Marker II and then processed in  
a Sakura VIP processor.  Two weeks ago the entire batch came out  
labeled much more lightly than usual, some to the point of where the  
label was completely effaced.
Our current theory is that an acid cleanser (Citronox) or possibly  
another acid was accidentally introduced into the processor.

Has any lab experienced this problem before, especially with Citronox?

Thank you.


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Fwd: [Histonet] (no subject)

[Malika Benatti]

Dear Histonet list manager

Is they anyway you could filter/block spam such as this one from be  
sent out to the Histonet list.


Cheers

Malika Benatti BSc MIBMS
Specialist Biomedical Scientist
Great Ormond Street Children Hospital
London

 " ... Smile it confuses people ..."

Begin forwarded message:


From: Green JumpyOne 
Date: 31 March 2010 09:52:42 GMT+01:00
To: , , >

Subject: [Histonet] (no subject)




http://www.trainedlabor.com/H6UH4Yh1UJ.html

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Re: [Histonet] Removing tissues from OCT for molecular analysis

[Malika Benatti]

What you should do ideally is sample you tissue for molecular studie  
first, then carry out frozen sections


Malika


 " ... Smile it confuses people ..."

On 29 Mar 2010, at 15:31, Katie Crosby  wrote:

I would like to remove tissues from blocks of OCT and perform  
molecule studies.  I need to separate tissues from OCT.

Thanks,
Katie

On Mar 29, 2010, at 10:21 AM, Malika Benatti wrote:


Do you mean removing OCT from tissue ?

" ... Smile it confuses people ..."

On 29 Mar 2010, at 13:55, Katie Crosby   
wrote:



Hello,

Can someone suggest a means of removing tissues from OCT for  
subsequent molecular analysis?  I did not find anything in the  
archives describing this exactly.


Thanks,

Katie






Katie Crosby
Immunohistochemistry
Cell Signaling Technology
3 Trask Lane
Danvers, MA 01923

Phone: 978-867-2352
Fax:  978-867-2400
WEB Site:   www.cellsignal.com

Toll Free:
1-877-616-CELL  (877-616-2355)
1-877-678-TECH  (877-678-8324)




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Katie Crosby
Immunohistochemistry
Cell Signaling Technology
3 Trask Lane
Danvers, MA 01923

Phone: 978-867-2352
Fax:  978-867-2400
WEB Site:   www.cellsignal.com

Toll Free:
1-877-616-CELL  (877-616-2355)
1-877-678-TECH  (877-678-8324)





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Re: [Histonet] histo techs doing cyto prep work

[Malika Benatti]

Working in Pediatric, without a cytology department, I get opportunity to 
handle non-gynae cytology sample such as Bronchial Alveolar Lavage, CSF, urine, 
cyst aspirate  blood sample, though all the sample are handle by Registered 
Biomedical Scientist that have been trained to handle cytological specimen. 


Malika 


Malika Benatti  

Specialist BMS
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH
United Kingdom


Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875




On 26 Mar 2010, at 16:42, Kim Tournear wrote:

> Hi all,
> I'm curious!! How many of you histotechs are being trained or already know 
> how to do cyto prep work? And how many of you histo supervisors are now 
> supervising cytology labs in addition to the histology lab? 
>  
> Is cross training histotechs to be cyto prep techs (in addition to their 
> histo job) becoming more popular?
>  
> I welcome any and all responses...
> 
> 
> 
> ~Kim Tournear ~ HT (ASCP), QIHC (ASCP)
> Histology Supervisor
> Tucson Medical Center
> Tucson,  AZ
> ~Don't let your life end before it begins~
> OU Rocks
> 
> 
> 
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Re: [Histonet] Paraffin Question

[Malika Benatti]

This will depend on the amount of blocks processed.

 I worked in places who used to change their VIP twice a week on Wednesday & 
Friday, and other that would change processor solutions once a week, but as a 
rule, processor wax regardless of the make should ALWAYS be changed after a 
maximum of 5 run for optimal result. 

Cheers,

Malika 

My current lab
On 26 Mar 2010, at 14:57, kristen arvidson wrote:

> How often are people changing/rotating their paraffin?  In other words the 
> dirtiest paraffin is how many days old?
> 
> 
> 
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Re: [Histonet] freezing mouse heart tissue

[Malika Benatti]

You may found that excessive amount of saline could cause freezing artefacts, 
so dabe

Try to embed heart sample on cryostat chuck directly, with OCT then freeze 
chuck on dry ice block directly as opposed to using liquid nitrogen.

Another option, would be freezing tissue in hexane boiling tube, with 
Acetone/dry ice freezing mixture. 

Hope this help.

Malika 



On 26 Mar 2010, at 13:22, Merced M Leiker wrote:

> Hi David,
> 
> I wouldn't snap-freeze the tissue in the OCT blocks. For one this could cause 
> the blocks to crack; you may have noticed this already in some of your blocks?
> 
> But you may want to try freezing the OCT block by resting the block in a tray 
> of isopentane (2-methylbutane) that has been frozen over liquid N, as you've 
> already thought of.
> 
> You can also search the Histonet archives on this topic as it's been 
> discussed before here.
> 
> Regards,
> Merced
> 
> 
> --On Friday, March 26, 2010 8:26 AM +0100 David Santer  wrote:
> 
>> Hello,
>> 
>> 
>> 
>> I am currently trying to produce cryosections from mouse heart tissue. I
>> already have experience with paraffin-sections and had faced no major
>> problems. But with cryosectioning I would ask for your help. You can get
>> an idea of our current status at
>>  this link  (Hematoxylin test
>> stain, not H&E, the whole heart section looks like this) and as you might
>> guess I am not satisfied with the quality. Would you call this freezing
>> artifacts? Some people suggested that freezing with only LN2 would be not
>> quick enough and create those ice crystals.
>> 
>> 
>> 
>> Here is how we prepared the tissue:
>> 
>> After taking out the hearts from the mice, we flushed them retrogradely
>> via the aorta with cold sodium solution. Then we cut the hearts in half,
>> put them into cryomolds and covered them with OCT. Afterwards they were
>> snap-frozen in liquid nitrogen and stored at -80°C.
>> 
>> 
>> 
>> Do you have an advice or maybe a suitable protocol for me? Would you
>> recommend 2-methyl butane?
>> 
>> 
>> 
>> Thank you very much! Greetings from the sunny Vienna!
>> 
>> 
>> 
>> David
>> 
>> --
>> 
>> Mit freundlichen Grüßen
>> 
>> with kind regards
>> 
>> 
>> 
>> Dr. David Santer
>> 
>> 
>> 
>> Ludwig Boltzmann Cluster for Cardiovascular Research
>> 
>> c/o Core Unit for Biomedical Research
>> 
>> Waehringer Guertel 18-20 - Leitstelle 1Q
>> 
>> A-1090 Vienna
>> 
>> Austria
>> 
>> 
>> 
>> Website:  
>> www.cardiovascular-research.at
>> 
>> 
>> 
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> 
> 
> 
> Merced M Leiker
> Research Technician III
> Cardiovascular Medicine
> 348 Biomedical Research Building
> State University of New York at Buffalo
> 3435 Main St, Buffalo, NY 14214  USA
> lei...@buffalo.edu
> 716-829-6118 (Ph)
> 716-829-2665 (Fx)
> 
> No trees were harmed in the sending of this email.
> However, many electrons were severely inconvenienced.
> 
> 
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Re: [Histonet] primate brain

[Malika Benatti]

At what thickness do you cut your sections ?

For Human brain we cut section at room temperature moist in Molifex and cut
sections at 7 µm  for standard HE/ 14 µm  for LFB.

Not sure what is the melting point of the wax your use, but if it is around
57 oC you can safely raise the temperature of your water bath to 50 oC.

Hope this help.

Malika

On Thu, Mar 25, 2010 at 9:17 PM, Beth A Gray  wrote:

> I have some primate brain blocks to do.  When I section the blocks the
> sections look nice.  When I put the ribbon on the water bath(  temp is 47)
> the tissue is wrinkled around the edges.  No amount of time on the water
> makes this better.  Ideas to solve this problem would be appreciated.
>  Thanks.
>
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Re: [Histonet] coverslipper

[Malika Benatti]

We use the Leica CV5030 the advantage of it is that it can be attached to
their Auto stainer providing continuous workflow as rack and coverslip 30
slides per run, without the need to physically transfer slides rack between
the autostainer and coverslipper or used as a stand alone coverslipper if
needed, though at time it can be problematic.

For a start glass cover slip must be kept at 37 oC prior use otherwise they
tend to stick to each others.
Also does not recognised all the slides type unless they are Leica one, or
the coverlslipper as been calibrated to use a specific slides type.

In the past I used the Sakura Acetate film coverslipper, and their were no
major problem with it though remounting section was a very tedious process.

Malika


On Thu, Mar 25, 2010 at 5:37 PM, Lynette Pavelich wrote:

> We've had our Sakura film coverslipper since '94 and love it.  The
> goodsaves us tech time coverslipping for hours each day, plus
> you can file the slides right away.  The bad.the docs prefer the
> bone marrow smears to be coverslipped by hand.  We have about 12 bone
> marrows a monthso reallyit's not a problem.  We've also
> tried other brands of film, but the cytotechs could notice a refractive
> difference, so we switched back to the Sakura brand.
>
> >>> "Margaryan, Naira"  3/25/2010
> 12:07 PM >>>
> Hi Colleges,
>
> I need your opinion about coverslip by hand vs. using machine.
> If you use machine what company's coverslipper you prefer?
>
> Thanks in advance,
> Naira
>
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Re: [Histonet] H & E QC

[Malika Benatti]

Hi there,

HE QC should be carry out on every slides that are stained, if you are checking 
for the staining intensity, this will be carried out using a control slide on 
new batch of staining solution, commercial or house made. Batch number should 
be recorded, and slides labelled with batch reference, date and file 
accordingly, the same goes for daily QC of routine work. Prior each morning run 
a control slide is run QC. 

HE QC goes beyond staining intensity, as a number of artefacts can be 
introduced and HE unacceptable ( Shatters, creases, folds, scores, sections cut 
too thick, scams  ... to name fews ) and therefore each slides stained should 
be QC by experience Staff prior been sent out to pathologist, this QC should be 
recorded and audited on a regular basis.

Furthermore over here in the UK every laboratory belong to the UK-NEQAS 
http://www.ukneqas.org.uk/  and external body that run a number of EQA Schemes 
for histopathology.

For HE EQA for each run the NEQAS will ask to select slides a number of HE 
slides that were produce at a randomly selected date. Slide will then be send 
externally and score by external accessor, and benchmark against all the 
participant in the scheme. If more score too low, lab will be flag and 
investigated. 

For those of you in the US, I would be curious to know if you have a similar 
system, and how they work.

Cheers,

Malika 

On 25 Mar 2010, at 03:22, WILLIAM DESALVO wrote:

> 
> Whether you are using an automated stainer or hand staining, run a control 
> slide and review before any patient samples are stained. I also suggest that 
> only start or endpoint QC is not enough and you should consider incorporating 
> continuous QC/QA at regular intervals for the stain set-up, to ensure the 
> highest quality and provide adequate control of the process. You should be 
> able to determine, in a very short time, the end point of the stain set up 
> and then add QC checks for slide quality at 1/3 and 2/3 through the run or 
> anytime a solution container is changed or rotated.
> 
> 
> 
> In our lab, with the regents used and staining protocols available to select, 
> we have determined that a stain set of solutions, on our automated 
> instrument, will maintain agreed and desired quality the pathologist will 
> accept for 1500 slides (I strongly suggest counting slides, not runs or 
> racks). We stop processing patient slides and run the control slide at runs 
> 1, 500, 1000 (+- 10% to allow for process flow and variance). The slides are 
> reviewed for acceptance or rejection by a Coordinator or higher and when 
> acceptable,patient slides may be placed on the instrument. All QC slides are 
> saved for review and the QC maintenance sheet is filed daily. This process 
> captures the employee that set up and monitors the instrument and the 
> employee that QC'd along w/ the QC review results. The control slide is a 
> multi-tissue slide that must contain the four highest volume tissue types for 
> the lab. Each pathologist receives a daily Quality Review sheet to report any 
> variance, issues or problems for all cases read.
> 
> 
> 
> Think through your process, communicate with the pathologist and develop a 
> QC/QA system/process that creates accountability for all employees, supports 
> production of quality results and meets your regulatory needs.  
> 
> William DeSalvo, B.S., HTL(ASCP)
> System Production Manager
> 
> Sonora Quest Laboratories
> 
> NSH Quality Control Committee Chairperson
> 
> 
>> From: adesupo2...@hotmail.com
>> To: histonet@lists.utsouthwestern.edu
>> Date: Wed, 24 Mar 2010 17:53:12 -0400
>> Subject: [Histonet] H & E QC
>>> 
>> Hi,
>> 
>> I will appreciate it, if you guys could share your method/procedure for H & 
>> E QC with me. Thanking you all for your usual cooperation.
>> 
>> 
>> 
>> Adesupo A.
>> 
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[Histonet] Contamination..processor?

[Malika Benatti]

Hey Brandi,

In my experience carry over generally occurred at time of cut-up or embedding 
when tiny bit of tissue get stuck between the groove of the forceps, despite 
been wiped clean between each specimens, but if you are 100 % sure that carry 
over did not occurred at time of cut-up, or embedding but during processing but 
during processing and the tonsil tissue is definitely in the block I suggest 
that you use bio-wrap/tissue wrap. 
Cheers,

Malika


Malika Benatti 

Specialist BMS
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH
United Kingdom


Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875

ben...@gosh.nhs.uk



Hello everyone.

 Today we have a problem with contamination.  The pathologist notes cells 
from tonsil specimens here and there on our GI biopsy slides.  The cells are in 
the block.  I'm trying to ascertain the source of the contamination.  
 The grossing pathologist grossed the tonsils AFTER all GI specimens 
yesterday (not source of contaminant).  We (the techs) embedded all GI 
specimens first, trimmed, cut, floated and stained ALL GI specimens BEFORE the 
tonsils (not source of contaminant).  The only other source of the 
contamination I can think of is from the tissue processor.  We have a Tissue 
Tek VIP closed processor.  Has anyone ever experienced any problems like this?  
We had a similar issue a few weeks ago.  I thought the contaminant cells may be 
from a bladder tumor, which had multiple sections submitted.  In this instance 
the cells showed up days work of the bladder tumor, and in the following days 
work also (though the pathologists could not say for sure the cells were from 
the bladder case).  We changed our formalin solutions in the processor and the 
problem did not present the next day.  We also started putting all bladder 
tumor specimens in the microcassettes, to prevent tissue from escaping.  Has 
anyone had any problem like this, or does anyone have any ideas on how to 
prevent this in the future?  We had not seen this problem until these past two 
incidences, and this tonsil problem is particularly strange to me because we 
process tonsils and GI specimens in the same workload on a regular basis and 
have never had this issue before.  Any help is appreciated!  

Thanks!
Brandi

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Re: [Histonet] Alcian Blue 2.5

[Malika Benatti]

Hey Laurie,

Try with home made Alcian Blue Solution : pH 2.5 1g Alcian Blue in 100  
ml of 3% Acetic Acid  ( check pH adjust accordingly, filter AB  
solution before use) also rather than rinsing with large amount of  
water, blot dry section this should help alteration of staining, then  
rinse briefly before next step.


Hope this help

Malika

Malika Benatti

Specialist BMS
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH


Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875

ben...@gosh.nhs.uk

 " ... Smile it confuses people ..."

On 24 Mar 2010, at 09:32, "Laurie Elmgren"   
wrote:



Hi Everyone,

Lately, we have been having a problem with our Alcian Blue. It is fine
up to the washing step. Once it hits the water, the blue fades to
unacceptable. We have tried reducing the water rinse after the blue to
just a quick rinse in d-H20, and it is still poor. The vendor replaced
the kit, and it worked fine the first time, then two weeks later, the
same problem arose. Three different techs have tried the stain three
times. Any suggestions?



Laurie Elmgren

Histology Supervisor

Sunrise Medical Labs

240 Motor Pkwy

Hauppauge, NY 11788

(631)435-1515x1108





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intended recipient of this message you must not disseminate, copy or
take any action in reliance on it."





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Re: [Histonet] number of slides

[Malika Benatti]

Routinely

Gastric bx  upper / rt colon / lt colon one rubbon of 4 sections per slides
Liver HE @ level 1/2/3 and liver specials, AE1 IHC
Renal Biopsy (Native and Transplant) 24 slides inc HE/Renal Special/ IHC and
USS
TransBronchial BX HE @ level 1/2/3 and lung special
BMT HE, retic and IHC for NBLX case
Endomyocardial BX, HE @level 1/2/3 EVG/MT and C4d IHC
Vascular Malformation HE and IHC
Surgical Heart (11 blocks) HE, EVG on all Blocks
Cornea Button HE, PAS, AB, Congo red
All Tumour BX HE, and 10 USS for IHC
PM Brain HE/LFB on all blocks
PM Heart HE/EVG on all blocks
PM Lung HE/EVG/PERLS on all blocks

Working in paediatric I have not seen a prostate of a breast for the past 6
years, but my old lab had between 4 and 6 blocks per prostate or breast
tissue and were cutting HE at level 1/2/3 and picking up 4 USS between level
for further IHC.


Malika

Malika Benatti
Specialist Biomedical Scientist
Great Ormond Street Hospital for Children


On Tue, Mar 23, 2010 at 10:08 PM, Sue  wrote:

> We do three levels on all diagnostic biopsies. Liver and Kidney also get
> upfront special stains, as do gastric biopsies (h pylori)
> We did cut down on extra unstained slides since we were discarding most of
> them. As far as stopping levels, my pathologist thinks it goes against
> standard
> of care. As for prostate biopsies, they are so small any more, that we are
> thinking o cutting 4 slides staining 1and 4 for H&E and holding 2 and 3 for
> possible IHC. We
> are finding that when we have to go back there is minimal tumor for IHC
> demonstration.
>
> Susan T. Paturzo
> Thomas Jefferson University Hospital
>
>
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Re: [Histonet] US HT

[Malika Benatti]

Janelle

The HPC registration rules have changed but most of the info you need are
available on the IMBS website
http://www.ibms.org/ and the HPC website http://www.hpc-uk.org/

If you have a recognised Degree it should only be a matter of getting your
Part One the of the HPC Registration to be able to be employed has a
registered BMS, then with the Part 2 you get your speciality in the
discipline of your choice.


Hope this help.

Malika



On Tue, Mar 23, 2010 at 9:59 PM, Janelle Powers
wrote:

> Hello Malika,
> I am not sure about sponsorship and immigration but what I can tell you is
> that to apply for an HT from ASCP you can take the exam which requires you
> to have either an associates degree with a combined twelve credits in
> chemistry and biology, as well as one year experience as a histotech, or you
> can take a NACCLE'S accredited course to be eligible for the exam. You maybe
> able to convert you UK credentials to ASCP I am not totally sure. This
> website should help
> http://www.ascp.org/FunctionalNavigation/certification/International.aspx.
> Hope this helps a little. I myself wanted to know what the requirements
> would be  if I wanted to convert my HT ASCP into UK accreditation. Any
> advice you have would be great! Thanks and good luck!
>
> Janelle
>
>
>
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Re: [Histonet] Reticulin Stain

[Malika Benatti]

Hi Alison,

Do you have a problem with all type of tissue ( Liver, Lung, Lymph Nodes,
Trephine ... ) or just Trephine specimens. ?

If you only have problems with Retic on BMT, if you use Formic Formaldehyde
Decal fluid, see if EDTA make a difference.

If you experience issue with all type of tissue. The ammoniacal silver may
be at fault, make a new batch and test it with liver or reactive Lymph Node
hopefully it will work.


In my lab we use the Gordon & Sweet Retic.  Staining protocol copy can be
found in any copy of Bancroft & Stevens or the newer version Bancroft &
Gamble, and Bancroft & Cook if you don'g have access to any of these book
you can download the staining protocol GORDON & SWEET'S
RETIC<http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF>


Hope this help.

Malika

Malika Benatti

Specialist BMS
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH
United Kingdom


Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875

ben...@gosh.nhs.uk


On Tue, Mar 23, 2010 at 8:35 PM, Scott, Allison D <
allison_sc...@hchd.tmc.edu> wrote:

> Hello to all in histoland.  We are having a problem with our reticulin
> stain.  It is not showing a real delination of the reticulin fibers.
> They are there but it tends to fade off.We do the brown and hopps
> stain.  Any help would be appreciated.
>
> Allison Scott HT(ASCP)
> Histology Supervisor
> LBJ Hospital
> 5656 Kelley
> Houston, Texas 77026
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Re: [Histonet] UK Trained and fully HPC Registered Histotechnologist looking to relocate in the US

[Malika Benatti]

Hi Bernice,

Thanks for your reply and the URL,  I hold a BSc Hons in Biomedical Sciences 
and a Post Graduate Certificate in Cellular Pathology over here in the UK.

Since you mentioned in your reply that you have a Histotechnologist who trained 
in Romania, I would be interested to know if she apply for her job from outside 
the US or was she already in the US when she applied for her position.

The problem that I face is although Histotechnologist are a "rare bread of 
Scientist" where automation has it's limitation and therefore very much in 
demand. Yet every time that I apply for a job, applying from outside the US, I 
find it very difficult to fill online pre-formatted application form, as they 
are designed for US based applicant. I have also sent my resume to few lab that 
have advertised job but, never got a feed back of any kind. 

Also Applying from abroad, their is the sponsorship/visa issue. 
I need a job to get working VISA a right now I feel like I am in a no win 
situation.  
Do you, or anyone on the histonet list reading this message who know of any 
recruiting company, laboratory manager, institutions in the US who are willing 
to deal with applicant from outside the US.

Regards, 

Malika




On 22 Mar 2010, at 14:51, Bernice Frederick wrote:

> Malika,
> ASCP has a list of accreditaion agencies here. You can send your transcripts
> to them (for a fee of course) and they will tell you if you need any other
> classes etc and what your scholastic status is. We have a tech from Romania
> and there she had a CLS degree. The accreditation agency gave her the
> equivalent of a BS in Medical Technology. She can take the HTL and plans to
> in May. The ASCP has a study Guide and the NSH has a whole series of self
> assessment books. It can be done.
> If you are here for the NSH meeting in September it would help you a lot as
> they have a class for persons taking the exam etc. Look at www.NSH.org.
> www.ascp.org is the ascp site and there is info on acceeditaion there.
> Bernice
> 
> 
> 
> Bernice Frederick HTL (ASCP)
> Northwestern University
> Pathology Core Facility
> ECOGPCO-RL 
> 710 N Fairbanks Court
> Olson 8-421
> Chicago,IL 60611
> 312-503-3723
> 
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Malika
> Benatti
> Sent: Sunday, March 21, 2010 5:12 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] UK Trained and fully HPC Registered
> Histotechnologistlooking to relocate in the US
> 
> To all Histotechnologist States Side,
> 
> I am fully qualified in the UK in all aspect of the work in histology with 8
> years post UK Health Professional Council (HPC) Registration (2 years in
> General Pathology, 6 years in Pediatric Pathology). 
> 
> I was wondering if someone on the Histonet list has ever sponsored/employed
> a Histotechologist who trained in the UK and then went to work in the US.  I
> am currently looking to relocate to the US and would like to inquire as to
> how I can convert my UK HPC registration into a US ASCP Certification, so
> that I may qualify to apply for histotechnologist jobs in the US. 
> 
> Kind regards,
> 
> Malika___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 


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[Histonet] UK Trained and fully HPC Registered Histotechnologist looking to relocate in the US

[Malika Benatti]

To all Histotechnologist State Side,

I am fully qualified in the UK in all aspect of the work in histology with 8 
years post UK Health Professional Council (HPC) Registration (2 years in 
General Pathology, 6 years in Pediatric Pathology). 

I was wondering if someone on the Histonet list has ever sponsored/employed  a 
Histotechologist who trained in the UK and then went to work in the US.  I am 
currently looking to relocate to the US and would like to inquire as to how I 
can convert my UK HPC registration into a US ASCP Certification, so that I may 
qualify to apply for histotechnologist jobs in the US. 

Kind regards,

Malika___
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