[Histonet] Histology Training Opportunities in KY?

Billing Consultants, LLC billingconsultants <@t> yahoo.com
Sat Feb 26 23:05:42 CST 2005

Hi everyone,
Would any of you know of any training opportunities in Kentucky for someone interested in becoming a histology technician?  I have a friend in Lexington that has a BS in biology and is really interested in breaking into the field.  She would love to find a place - even volunteer her time if necessary for the chance to learn.  
Any suggestions you may have would be greatly appreciated.  
Louri Roberts-Caldwell

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Today's Topics:

1. magnesium detection (Kristopher Kalleberg)
2. rescue sections for staining (J. Douglas Swarts)
3. Equipment (David Deibler)
4. 35BH11 (Sebree Linda A.)
5. paraffin embedding of mouse tails (Cheung, Lauren)
6. RE: stent techniques anyone??? (Linda Jenkins)
7. GFP (Carol Bobrowitz)
8. CXCR4 & CXCL12 antibodies ( Judy Strauss)
9. Re: richard-allen type 9 paraffin (TheBestTime23 <@t> aol.com)
10. Re:35BH11 (Linda Davis)
11. Fw: Re:35BH11 (Linda Davis)
12. Histotech Marketability (cfockler <@t> mail1.vcu.edu)
13. Square "sample stage" (HIRANO Makoto)
14. RE: richard-allen type 9 paraffin (Hugh Luk)
15. RE: Histotech Marketability (Luck, Greg D.)
16. HT vs. HLT (cfockler <@t> mail1.vcu.edu)
17. Collagen-I gel, dye and autofluorescence (Tim Demuth)


Message: 1
Date: Fri, 25 Feb 2005 13:34:23 -0500
From: "Kristopher Kalleberg" 
Subject: [Histonet] magnesium detection
To: "histonet <@t> lists.utsouthwestern.edu"



Does anyone know of a way to detect magnesium or calcium, through IHC or
fluorescence, in skin? What about quinalizarin? THanks.

Kris L Kalleberg
Unilever R&D
45 River Rd. 
Edgewater, NJ 07020
201 840 2472


Message: 2
Date: Fri, 25 Feb 2005 13:35:48 -0500
From: "J. Douglas Swarts" 
Subject: [Histonet] rescue sections for staining
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <421F7004.5010509 <@t> pitt.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

We are trying to stain some sections cut about ten years ago. They are 
rat cranial bases including the middle ear which is the area we are 
interested in. When we stain the sections with H&E we get no 
hematoxylin staining, ie. no nuclei at all, but the eosin gives a nice 
uniform pink color. 

The specimens were decalcified in 10% formic acid which was neutralized 
with sodium sulfate according to the histologists notes. The sections 
stained at the time show very little hematoxylin staining. So the 
questions are:

Are these sections overdecalcified? If so is there any technique I can 
use to rescue them?

Did the neutralization with sodium sulfate create the problem? If so 
can I do anything to reverse the effect?

Finally, if one or the other of these situations are true and cannot be 
remedied is there another staining protocol that will give me a 
"reasonable" level of morphologic detail?

Thanks for your help.



Message: 3
Date: Fri, 25 Feb 2005 12:47:43 -0600
From: "David Deibler" 
Subject: [Histonet] Equipment
Message-ID: <000801c51b6a$7e380110$2f01010a <@t> tamtron.ctpl.dns>
Content-Type: text/plain; charset="iso-8859-1"

We are looking for an old Shandon portable coverslipping 
hood. Our lab needs to buy on for IHC. If anyone has a 
used one let me know , we would love to purchase it.

David Deibler
ddeibler <@t> centexpathlab.com


Message: 4
Date: Fri, 25 Feb 2005 13:16:22 -0600
From: "Sebree Linda A." 
Subject: [Histonet] 35BH11
To: "Histonet \(E-mail\)" 

Content-Type: text/plain; charset="iso-8859-1"

Hi all,

I'm wondering if there is a reference lab out there (preferably offering "stain only") that does 35BH11 stain. I'm assuming that's a clone but I don't have any other name for it.


Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
600 Highland Ave.
Madison, WI 53792
FAX: (608)262-7174


Message: 5
Date: Fri, 25 Feb 2005 11:14:52 -0800
From: "Cheung, Lauren" 
Subject: [Histonet] paraffin embedding of mouse tails
To: "'histonet <@t> lists.utsouthwestern.edu'"

Message-ID: <8022E9EE6D5ED511A39200A0C9DED0E902FF358E <@t> cvexchange>
Content-Type: text/plain; charset="iso-8859-1"

Hi everyone,

I work with mouse tail sections, each approximately 4-5 mm in thickness. I
have been attempting to embed them in paraffin for sectioning however
sectioning them hasn't been very successful. I can't obtain a ribbon and the
sections seem to break out of the paraffin and break apart. I currently fix
the tails in 4% Paraformaldehyde overnight and then place them in EDTA for
7-10 days for decalcification. Afterwards, I dehydrate them through a series
of graded alcohol steps, clear them in xylenes, and then let them sit in
paraffin. I am wondering if anyone has a good protocol including times for
each step which I could test out on my samples. Any advice would be greatly

Thanks so much,

Lauren Cheung
Stanford University
Rockson Lab
Lab: 650-723-5354
Email: Lcheung <@t> cvmed.stanford.edu


Message: 6
Date: Fri, 25 Feb 2005 14:48:30 -0500
From: Linda Jenkins 
Subject: [Histonet] RE: stent techniques anyone???
To: histonet <@t> lists.utsouthwestern.edu
< <@t> mailhost.ces.clemson.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

You stated:
"ok...here is one where i need some guidance.....post mortem manangement of
coronary artery stents....those little metal mesh things...should i push the
clot out and section it??
gil corrigan msmdphd tigergil <@t> aol.com best practise anyone???
ps we are starting a computer section in the aafs ...in case you known some
forensic nurds...send them to tigergil <@t> aol.com"

Removing the stent or clot can remove a lot of valuable 
information regarding biocompatibility. A better choice would be to embed 
the section in plastic (methyl methacrylate, glycol methacrylate, etc.) and 
then section on an automated microtome with tungsten carbide blade or slab 
section with a diamond coated blade and grind the sample to the desired 
thinness. Of course, this requires specialized equipment and technical 
skill that many routine clinical labs may not have. You might want to 
consider sending these out for contract work.
The private contract labs that process stents on a routine basis 
have some really good procedures BUT cannot (unfortunately) share with 
others as they are bound by non-disclosure agreements.
If you are familiar with plastics, you may want to give it a try!
Good Luck,

Linda Jenkins, HT
Clemson University
Dept. of Bioengineering
Clemson, SC 29634-0905


Message: 7
Date: Fri, 25 Feb 2005 14:21:06 -0600
From: Carol Bobrowitz 
Subject: [Histonet] GFP
To: "'Histonet (E-mail)" 
Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9640 <@t> thor.phys.mcw.edu>
Content-Type: text/plain; charset="iso-8859-1"

Has anyone worked with GFP tissue?

I have found GFP is surviving frozen sections, acetone and 100% alcohol. 

I am wondering about FFPE tissue. I'm thinking the heat during processing
would destroy the GFP.

Does formalin fixation effect the GFP?

Is it possible to fix the tissue in an alcoholic fixative and manually hand
process at room temperature and infiltrate into plastic? Would the plastic
medium destroy the GFP?

What about celloidin embedded tissue? This is not my choice

Any information would be helpful.

I thank everyone on the histonet that has in the past and future helped me.
I really appreciate your help.


Carol Ann Bobrowitz
Histology Laboratory
Department of Physiology - Room 541
Medical College of Wisconsin
8701 Watertown Plank Road
Milwaukee, Wisconsin 53226
FAX 414-456-6546
cbobrowi <@t> mcw.edu


Message: 8
Date: Fri, 25 Feb 2005 15:39:58 -0500
From: " Judy Strauss" 
Subject: [Histonet] CXCR4 & CXCL12 antibodies
Content-Type: text/plain; charset=US-ASCII

Can anyone recommend antibodies to CXCR4 (fusin) and CXCL12 that are
suitable for immunohistochemistry in formalin fixed paraffin embedded rat


Judith Strauss
SUNY Upstate Medical University
Department of Orthopedic Surgery
IHP room 3118
505 Irving Avenue
Syracuse, NY 13120

phone: (315) 464-9960
fax: (315) 464-6638


Message: 9
Date: Fri, 25 Feb 2005 16:43:50 EST
From: TheBestTime23 <@t> aol.com
Subject: Re: [Histonet] richard-allen type 9 paraffin
To: liz <@t> premierlab.com
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <92.2169ca98.2f50f616 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"

Thanks for your reply. How long have you been using this paraffin? What 
temperatures are you using in your water baths and processors? What do you use 
to keep your blocks chilled (if anything)?

I'm having a little difficulty with this new paraffin. I thought it was 
something that I needed time getting used to, but it has been almost 2 months of 
using it and I am still having problems (and so are the 3 other people I 
work with). Problems include compression, fragile ribbons and wrinkles. I am 
now taking an excessive amount of time and care cutting my blocks only to 
discover minute folds that could not be seen on the waterbath. We cut our 
routine sections at 4 microns and our temperatures are where they should be for 
what the company recommends (59 in processor and holding tanks and waterbath 
between 35-40). I just thought someone that had worked with this paraffin for a 
while could give me some tips. Otherwise I might have to start wearing a 
wig : )

Thank you!


Message: 10
Date: Fri, 25 Feb 2005 16:00:17 -0600
From: "Linda Davis" 

Subject: [Histonet] Re:35BH11
Message-ID: <006001c51b85$642517c0$de0ac942 <@t> D6JLZ851>
Content-Type: text/plain; charset="iso-8859-1"


Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain.

Linda Davis
Rio Grande Regional Hospital
McAllen, TX.
(956) 632-6402
Fax (956) 632-6641
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Message: 11
Date: Fri, 25 Feb 2005 16:22:53 -0600
From: "Linda Davis" 

Subject: [Histonet] Fw: Re:35BH11
Message-ID: <004001c51b88$8c565ee0$6d0ac942 <@t> D6JLZ851>
Content-Type: text/plain; charset="iso-8859-1"


Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain.

Linda Davis
Rio Grande Regional Hospital
McAllen, TX.
(956) 632-6402
Fax (956) 632-6641
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Message: 12
Date: Fri, 25 Feb 2005 17:55:56 -0500
Subject: [Histonet] Histotech Marketability
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <200502252255.RAA22846 <@t> arrakis.vcu.edu>
Content-Type: text/plain; charset=iso-8859-1

Hello histonetters. . .just a quick question. What is the
marketability (pay scale ect.) of a tech with a BS in
biology/chemistry, 2 years of experience with proven competency in all
aspects, grosses, and has IHC and IF experience? I am looking to get
feedback from all over the country. Any information would be greatly

Candyce Fockler
Anatomic Pathology


Message: 13
Date: Fri, 25 Feb 2005 17:48:01 -0600
From: "HIRANO Makoto" 
Subject: [Histonet] Square "sample stage"
Content-Type: text/plain; charset="iso-8859-1"

I am a beginner in frozen sectioning.
Now I have a silly question.

What is the formal name of a metal, square "sample stage" or "chunk" for
I mean the 'partner' of a cryomold.
I have visited some web sites and consulted some catalogues, but I cannot
find out the name.
I have absolutely to order some.

Thanks a lot in advance.

Department of Molecular Biosciences
University of Kansas
E-mail: mhirano <@t> ku.edu


Message: 14
Date: Fri, 25 Feb 2005 17:45:10 -1000
From: "Hugh Luk" 
Subject: RE: [Histonet] richard-allen type 9 paraffin
To: TheBestTime23 <@t> aol.com
Cc: histonet <@t> lists.utsouthwestern.edu
Content-Type: text/plain; format=flowed


Just my 2 cents. If any of this helps, great.

I've found this is one of the hardest, polymer-rich paraffins available. I 
find it similar to Surgipath's standard embedding paraffin and Cardinal's 
(old S/P) Ameraffin. It sections and ribbons well. Currently, however, I am 
using Ameraffin (also made by Richard Allan), which I find very 
interchangable with type 9.

We modified our tissue processing (a little more time in wax-multiple 
changes totalling around 2 hours at 58 to 62C- and maybe more alcohol), 
deparaffinization (more time in solvent), and two, different microtome 
blades. Sakura's Accu-Edge HIGH profile blades are honed perfectly to cut 
ANYTHING at 4 microns with this wax. The "Slight compression" is balanced 
with "Ease of ribboning." We sometimes use a wider angle on the blade 
holder to increase the rake angle.

For really thin sections, "Sharper" blades (like Duraedge, Leica's, or 
Richard Allan's blades) are required. For example, on delicate biopsies and 
lymph nodes, 1 or 2 microns is no problem! I also like the way the paraffin 
holds its shape on the water bath (~38-44C).

I learned histology using Paraplast X-tra. I've tried Paraplast, Paraplast 
plus (with DMSO), Fisher Tissue Prep and Tissue Prep2 embedding medias. 
When we had a new supervisor at QMC, she changed the paraffin to a harder 
wax. We complained about it because it felt different, but I came to really 
like it. It became my standard where ever I work (thanks Jackie O').

If you are experimenting, you will need to do full switch-overs, every 
instrument using wax should be changed. Mixing paraffins of different 
grades is cheating the experiment.

My conclusion is they are all pretty good. The paraffin is only for 
support; and with a little tinkering, I like to think I can use any one of 
them at any time.

Hugh Luk, HTL (ASCP)
Former histopathology supervisor CLH
hluk <@t> crch.hawaii.edu
Cancer Research Center of Hawaii
1236 Lauhala Street, Room 316
Honolulu, HI 96813
Tel: (808) 440-5238
Fax: (808) 586-2982

>From: TheBestTime23 <@t> aol.com
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] richard-allen type 9 paraffin
>Date: Wed, 23 Feb 2005 17:42:03 EST
>Hello everyone,
>The lab that I work for recently decided to switch to Richard-Allen type 9
>paraffin. I was wondering if anyone has experience with this paraffin and
>could give some advice on techniques used in cutting it. Opinions of this
>product would also be greatly appreciated.
>Thanks in advance,
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu


Message: 15
Date: Fri, 25 Feb 2005 23:53:17 -0800
From: "Luck, Greg D." 
Subject: RE: [Histonet] Histotech Marketability
To: "'cfockler <@t> mail1.vcu.edu'" ,
histonet <@t> lists.utsouthwestern.edu

Content-Type: text/plain


Request that the HR dept of your current employer do a job code market
survey for your geographic area. After they tell you your salary is
equivalent to current market conditions ask them how long it would take to
fill a vacancy in your area. When they say "We don't know" ask them to try
to fill a position (real or not). Then inform them what a "histo'temp"
would cost ($57-60 hr.) to fill in while ???? There is a good deal of
upward pressure on histotech salaries (well overdue) and given a suitable
fit now is the time to look towards the future for new opportunities An
individual's concept of their marketability may differ substantially from
their employer's but this is limited only by their current work environment.
I know, blah, blah, blah. We have a full-time, M-F, day shift, histotech
position open. This is a seasoned dept looking towards the future to lay
the foundation of a new staff to take over (+/-5 yrs) as we retire. We have
a current opportunity for someone either interested in the eventual
department manager's role or as a traditional bench histotech. Click on the
web links that follow to get a snapshot of our institution and of Spokane:
www.deaconessmedicalcenter.org & www.visitspokane.com. 
Remember, "making a living" is not the same thing as "making a life".
Thanks, Greg

Greg Luck, BS, HT(ASCP)
Anatomic Pathology Supervisor
Deaconess Medical Center
800 W. 5th Ave
Spokane, WA 99204
Phone 509.473.7077
Fax 509.473.7133
luckg <@t> empirehealth.org

-----Original Message-----
From: cfockler <@t> mail1.vcu.edu [mailto:cfockler <@t> mail1.vcu.edu] 
Sent: Friday, February 25, 2005 2:56 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Histotech Marketability

Hello histonetters. . .just a quick question. What is the marketability
(pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of
experience with proven competency in all aspects, grosses, and has IHC and
IF experience? I am looking to get feedback from all over the country. Any
information would be greatly appreciated! 

Candyce Fockler
Anatomic Pathology

Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu


Message: 16
Date: Sat, 26 Feb 2005 09:17:23 -0500
Subject: [Histonet] HT vs. HLT
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <200502261417.JAA24219 <@t> despina.vcu.edu>
Content-Type: text/plain; charset=iso-8859-1

What exactly is the difference between and HT and an HLT. ASCP makes
this distinction with regards to their wage and salary survey. Is one
certified and the other not? Any information would be greatly
appreciated! Thanks.


Message: 17
Date: Sat, 26 Feb 2005 08:52:48 -0700
From: Tim Demuth 
Subject: [Histonet] Collagen-I gel, dye and autofluorescence
Content-Type: text/plain; charset="ISO-8859-1"

Hi there,

we are currently trying to cut FFPE sections of multicellular
tumor-spheroids grown in collagen-I gel (bovine). There are two major
1) Since the spheroid is so small (diameter ~200-500µm) and very pale it is
extremely hard to identify in the fixed and embedded collagen block. We
would like to use a dye to visualize the spheroid prior to embedding, so we
have some guidance as to where and when to expect the spheroid so we don't

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