[Histonet] RE: Histonet Digest, Vol 54, Issue 21

Fri May 16 10:27:06 CDT 2008

I have not, as of yet, to see any delays or shortfalls of hematoxylin at this time in my orders.

Mike

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Featherstone, Annette
Sent: Friday, May 16, 2008 9:07 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Histonet Digest, Vol 54, Issue 21

What is everyone doing about the hematoxylin shortage? 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Thursday, May 15, 2008 12:48
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 54, Issue 21

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Today's Topics:

   1. Glucose oxidase method for blocking endogenous peroxidase
      (Gale, Nadia)
   2. Cutting research blocks from home? (Pam Bakken)
   3. Re: ZYMED Histostain Plus Kit on animal tissue (Kim Merriam)
   4. Re: Cutting research blocks from home? (Rene J Buesa)
   5. Re: Salary Scales (Rene J Buesa)
   6. Suggestions on Equipment?? (MICHELLE SEAGLE)
   7. Re: Suggestions on Equipment?? (TBritten <@t> aol.com)
   8. cd38 on mouse tissue (Patsy Ruegg)
   9. Re: Suggestions on Equipment?? (Rene J Buesa)
  10. Sending privately Re: [Histonet] Glucose oxidase method for
      blocking	endogenous peroxidase (Gayle Callis)
  11. vibratome sections for cavalieri volume estimation (Claudia Lutz)
  12. Re: ZYMED Histostain Plus Kit on animal tissue (Gayle Callis)
  13. OCT vs. isopentane (shupeiwu <@t> umich.edu)
  14. Re: Standardized Microtomes (Amos Brooks)
  15. RE: OCT vs. isopentane (Monfils, Paul)
  16. Re: Standardized Microtomes (Gayle Callis)
  17. Help identifying LS174T tumor cells (jstaruk)
  18. Gamal Akabani has sent you a hi5 Friend Request (Gamal Akabani)
  19. Re: Help identifying LS174T tumor cells (koellingr <@t> comcast.net)
  20. FW: [Histonet] Help identifying LS174T tumor cells (Jim Manavis)
  21. Re: Suggestions on Equipment?? (Anne van Binsbergen)
  22. Re: Standardized Microtomes (Rene J Buesa)
  23. Looking for Melissa Barrett (Whitaker, Bonnie)
  24. need knife guard (Rutledge, Nancy)
  25. RE: Salary Scales (Dawson, Glen)
  26. RE: Salary Scales (Rene J Buesa)
  27. Alternative Preserving Medium for Histology (Rupert Street)

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

Message: 1
Date: Wed, 14 May 2008 10:05:39 -0700
From: "Gale, Nadia" <Ngale <@t> bccancer.bc.ca>
Subject: [Histonet] Glucose oxidase method for blocking endogenous
	peroxidase
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<E76535B27E4D5A428863F402EC2E0C2901E6C600 <@t> srvex03.phsabc.ehcnet.ca>
Content-Type: text/plain;	charset="us-ascii"

I've searched the archives and cannot find what I am after... I'd like to try the glucose oxidase method for blocking endogenous peroxidase in tissue sections for IHC.

What is a suitable replacement for the B-d(+)glucose G5250 discontinued by Sigma?  Will the D-(+)-Glucose G8270 work?  This is 4% beta isomer and 96% alpha.

Thanks for your help,
Nadia

Nadia Gale
Lead Histotechnologist
Centre for Translational and Applied Genomics (CTAG) 3427-600 West 10th Avenue Vancouver, BC V5Z 4E6 tel 604-877-6000 ext 2426 fax 604-877-6089 ngale <@t> bccancer.bc.ca

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

Message: 2
Date: Wed, 14 May 2008 12:39:57 -0500
From: "Pam Bakken" <Pam.Bakken <@t> childrensmn.org>
Subject: [Histonet] Cutting research blocks from home?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <482ADD9D.37F9.0000.0 <@t> childrensmn.org>
Content-Type: text/plain; charset="us-ascii"

Hi,

I am interested in working part time from home cutting research blocks.  Is there anyone that could give me some advice on how to get started? How do I charge?  Any experience, advice or knowledge would be greatly appreciated.  

Thanks,

Pamm Bakken
Children's Hospital - Minneapolis _______________________________________________________________

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Message: 3
Date: Wed, 14 May 2008 10:51:11 -0700 (PDT)
From: Kim Merriam <kmerriam2003 <@t> yahoo.com>
Subject: Re: [Histonet] ZYMED Histostain Plus Kit on animal tissue
To: Mary L Giebel/FS/VCU <mlgiebel <@t> vcu.edu>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <654253.27119.qm <@t> web50308.mail.re2.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

I dont have experience with this particular kit from Zymed, but the term "broad spectrum" makes me think that it would be similar to a universal secondary (that might include anti-mouse and anti-rabbit secondaries) which would likely cross-react to your rat tissue.  It might be worth giving Zymed a call to find out about this kit.
 Kim Merriam, MA, HT(ASCP)
Cambridge, MA

----- Original Message ----
From: Mary L Giebel/FS/VCU <mlgiebel <@t> vcu.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Wednesday, May 14, 2008 12:56:51 PM
Subject: [Histonet] ZYMED Histostain Plus Kit on animal tissue

At a client's request, I am using a ZYMED's Broad Spectrum Histostain Plus Kit (#85-9843) on rat tissue. I have encountered an unexpected staining pattern as well as more background staining than usual. I would be very interested in hearing from others who have used this kit on animal tissue. I found the question posed in the Histonet archives (8/23/2005), but never found an answer posted. 
Thank you for your help.
Regards,
Mary Giebel

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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

Message: 4
Date: Wed, 14 May 2008 12:18:02 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Cutting research blocks from home?
To: Pam Bakken <Pam.Bakken <@t> childrensmn.org>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <734.60655.qm <@t> web65716.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

After getting the "essentials" (microtome, water bath, oven, staining dishes and needed supplies) you have to decide a price.
  Charge by the slide. One H&E stain costs about $4.50, with everything else (from cutting to staining) I would charge $10/stained section.
  René J.

Pam Bakken <Pam.Bakken <@t> childrensmn.org> wrote:

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

Message: 5
Date: Wed, 14 May 2008 12:21:40 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Salary Scales
To: JR R <rosenfeldtek <@t> hotmail.com>, histonet <@t> lists.utsouthwestern.edu
Message-ID: <623891.86971.qm <@t> web65712.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Which is equivalent to $26.20 / hour (2080 hours / year) and could be considered as "decent".
  René J.

JR R <rosenfeldtek <@t> hotmail.com> wrote:

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

Message: 6
Date: Wed, 14 May 2008 12:28:43 -0700 (PDT)
From: MICHELLE SEAGLE <mrsseagle <@t> yahoo.com>
Subject: [Histonet] Suggestions on Equipment??
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <739926.38110.qm <@t> web51803.mail.re2.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Our hospital is currently looking to purchase a new cryostat, H& E Stainer and an Embedding Center.  Any suggestions on a certain model,type or company with one that you have had a good experience with would be greatly appreciated.

  Michelle Seagle HT (ASCP)
  Rutherford Hospital 

MICHELLE SEAGLE

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

Message: 7
Date: Wed, 14 May 2008 15:36:57 EDT
From: TBritten <@t> aol.com
Subject: Re: [Histonet] Suggestions on Equipment??
To: mrsseagle <@t> yahoo.com, histonet <@t> lists.utsouthwestern.edu
Message-ID: <bc4.22039324.355c9959 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"

hello; we have used the "hacker" brand cryostat for many years at very high volumes and they have been very good. there are under a few brand marks but all  who sell such things will be able to help you identify the latest  model. 
tom

In a message dated 5/14/2008 3:30:34 P.M. Eastern Daylight Time, mrsseagle <@t> yahoo.com writes:

Our  hospital is currently looking to purchase a new cryostat, H& E Stainer and  an Embedding Center.  Any suggestions on a certain model,type or company with one that you have had a good experience with would be greatly appreciated.

Michelle Seagle HT (ASCP)
Rutherford Hospital 

MICHELLE SEAGLE

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

Message: 8
Date: Wed, 14 May 2008 14:32:05 -0600
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: [Histonet] cd38 on mouse tissue
To: "'Histonet'" <histonet <@t> pathology.swmed.edu>
Message-ID: <003301c8b601$9378f720$6401a8c0 <@t> Patsyoffice>
Content-Type: text/plain;	charset="us-ascii"

Anybody used cd38 on ffpe or frozen mouse tissue?

Thank you,

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net

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Message: 9
Date: Wed, 14 May 2008 13:40:16 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Suggestions on Equipment??
To: MICHELLE SEAGLE <mrsseagle <@t> yahoo.com>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <747728.50125.qm <@t> web65703.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Cryostat from Leica; H&E stainer and embedding center from Sakura, although it is very likely that the Leica people (that manufacture the 3 pieces of equipment) will give you a better price if you buy all from them. Otherwise follow the first choice (Leica + Sakura).
  René J.

MICHELLE SEAGLE <mrsseagle <@t> yahoo.com> wrote:

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

Message: 10
Date: Wed, 14 May 2008 15:46:23 -0600
From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
Subject: Sending privately Re: [Histonet] Glucose oxidase method for
	blocking	endogenous peroxidase
To: "Gale, Nadia" <Ngale <@t> bccancer.bc.ca>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000501c8b60b$f46ed9a0$6501a8c0 <@t> DHXTS541>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=original

Nadia,

I will attach the method and all the regeant needs to you privately and where you can purchase the correct glucose.  No, glucose D-(=) Glucose will NOT work.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT 59715
----- Original Message -----
From: "Gale, Nadia" <Ngale <@t> bccancer.bc.ca>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, May 14, 2008 11:05 AM
Subject: [Histonet] Glucose oxidase method for blocking endogenous peroxidase

I've searched the archives and cannot find what I am after... I'd like to try the glucose oxidase method for blocking endogenous peroxidase in tissue sections for IHC.

What is a suitable replacement for the B-d(+)glucose G5250 discontinued by Sigma?  Will the D-(+)-Glucose G8270 work?  This is 4% beta isomer and 96% alpha.

Thanks for your help,
Nadia

Nadia Gale
Lead Histotechnologist
Centre for Translational and Applied Genomics (CTAG) 3427-600 West 10th Avenue Vancouver, BC V5Z 4E6 tel 604-877-6000 ext 2426 fax 604-877-6089 ngale <@t> bccancer.bc.ca

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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

Message: 11
Date: Wed, 14 May 2008 17:12:59 -0500 (CDT)
From: "Claudia Lutz" <cclutz2 <@t> life.uiuc.edu>
Subject: [Histonet] vibratome sections for cavalieri volume estimation
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<4769.130.126.50.22.1210803202.squirrel <@t> www-s.life.uiuc.edu>
Content-Type: text/plain;charset=iso-8859-1

Hi,

I am fairly new to histological procedures and new to this list.  My lab would like to stain vibratome sections of honey bee brain with phalloidin in order to estimate volume of specific brain regions.  I am fixing the brains overnight in 4% paraformaldehyde, embedding in agarose, sectioning at 100 um, then staining free-floating sections in phalloidin with PBS with 0.2% Triton-X.  I am mounting the slices in 80% glycerol and then imaging with the confocal microscope.  I would like to use stereological procedures (specifically the cavalieri volume estimator) to quantify the volume of specific brain regions using systematic sampling of 10um optical sections that I obtain from the slices.  I have several (possibly naive)
questions:

1. My 100 um sections are shrinking to 60 um (or less) by the time I image them, judging by where I find or lose focus while imaging.  What can I do to minimize this?  How can I properly quantify the degree of shrinkage, to see whether or not it is uniform?  Can I avoid bias in my volume estimation?

2. My phalloidin staining appears much brighter in the center of the slice, even when I use settings to adjust the gain while taking z-stacks. 
Does this indicate tissue damage at the edges?  Can I ignore this?  This also relates to my question above, since I am not sure I can precisely find the edge of the slice by focusing when I need to increase the gain.

3. I lose order and orientation of the slices because I am staining them free-floating.  Does anyone have experience staining 40 or 50 um vibratome slices on the slide with phalloidin or antibody, or have a trustworthy protocol for this?

Sorry to ask so many questions, and thank you for any help.  There is not a lot of expertise in my lab on this, so I am feeling frustrated.

Claudia Lutz
University of Illinois at Urbana-Champaign Neuroscience Graduate Program

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

Message: 12
Date: Wed, 14 May 2008 16:22:26 -0600
From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
Subject: Re: [Histonet] ZYMED Histostain Plus Kit on animal tissue
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <003601c8b610$fd760000$6501a8c0 <@t> DHXTS541>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=original

Kim,

This kit detects (in its broad spectrum) - mouse, rabbit, rat and guinea pig.  You are correct about this more universal, multilink secondary.  The kit protocol is found on the Invitrogen website with information on what is in the secondary spectrum.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT 59715

----- Original Message -----
From: "Kim Merriam" <kmerriam2003 <@t> yahoo.com>
To: "Mary L Giebel/FS/VCU" <mlgiebel <@t> vcu.edu>; <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, May 14, 2008 11:51 AM
Subject: Re: [Histonet] ZYMED Histostain Plus Kit on animal tissue

I dont have experience with this particular kit from Zymed, but the term "broad spectrum" makes me think that it would be similar to a universal secondary (that might include anti-mouse and anti-rabbit secondaries) which would likely cross-react to your rat tissue. It might be worth giving Zymed a call to find out about this kit. Kim Merriam, MA, HT(ASCP) Cambridge, MA

----- Original Message ----
From: Mary L Giebel/FS/VCU <mlgiebel <@t> vcu.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Wednesday, May 14, 2008 12:56:51 PM
Subject: [Histonet] ZYMED Histostain Plus Kit on animal tissue

At a client's request, I am using a ZYMED's Broad Spectrum Histostain Plus Kit (#85-9843) on rat tissue. I have encountered an unexpected staining pattern as well as more background staining than usual. I would be very interested in hearing from others who have used this kit on animal tissue. I found the question posed in the Histonet archives (8/23/2005), but never found an answer posted. Thank you for your help. Regards, Mary Giebel

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

_______________________________________________
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------------------------------

Message: 13
Date: Wed, 14 May 2008 18:31:31 -0400
From: shupeiwu <@t> umich.edu
Subject: [Histonet] OCT vs. isopentane
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20080514183131.39132e3x7n46szfo <@t> web.mail.umich.edu>
Content-Type: text/plain;	charset=ISO-8859-1;	DelSp="Yes";
	format="flowed"

Hello all,

I am quite new in the field of histochemistry. I'm doing mouse intestine cryostat section right now. I'm so confused about the method for cryostat section.

First, I knew that most of time people using OCT to embed their sample, but I saw some papers they use liquid nitrogen cooled isopentane to embed their sample. Do these two methods cause any difference? Because I saw some background flurorescence in the OCT embedded slide.

And if the sample embedded in isopentane, does the cryostat section take place in the same kind of machine? (with -20oC cryosection)

Second, for the 4% Paraformaldehyde following by sucrose-PBS for cryoprotection, does it is really important to wash out the previous medium before entering the next step until the embedding.

Third, does the thickness of the slide affect the autofluorescence?

I knew those questions might some kind of vague.
But thank you very much for any answers ahead.

Wu

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

Message: 14
Date: Wed, 14 May 2008 18:42:54 -0400
From: "Amos Brooks" <amosbrooks <@t> gmail.com>
Subject: Re: [Histonet] Standardized Microtomes
To: "Rene J Buesa" <rjbuesa <@t> yahoo.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<582736990805141542i42937995ub1411468c2feda9b <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Sorry Rene,
     I absolutly disagree with that. The exact opposite: *Not* modifying the angle of your block holder to suit the block, is not good practice. You are bound to loose tissue when cutting archived blocks, or blocks from other labs or even slight variations within your own lab. If you don't adapt to the blockyou are cutting. Without modifying the angle of the holder, one has 2 choices: Just cut thru it (refacing) or re-embedding and thereby needing to re-trim. Both scenerios will loose tissue.
     Often I have cut a freshly embedded block where the cassette didn't sit perfectly flat on the mold, otherwise the embedding was perfect. (Often due to poorly dissected tissue with high points). This results in an angle that is slightly askew. Adjusting the block holder is much faster than re-embedding the block. If it is done properly you can actually modify the block angle without ever cutting any tissue even on a previously cut block.

We're going to have to disagree on this, Amos

On Wed, May 14, 2008 at 9:14 AM, Rene J Buesa <rjbuesa <@t> yahoo.com> wrote:

> All in all the practice of "changing the angle to match the block you
> have to recut as needed" is not a good practice.
> During it you could end loosing tissue when trying to "build" a new 
> flat surface from a block that was originally cut at a different 
> angle, while "sculping" the new surface.
> The best practice is to have all the microtomes cutting at the same 
> angle and try to convince those HTs that say that "I have always cut 
> with this angle" that they are flat wrong and a general angle can be 
> good for the majority of blocks.
> René J.
>

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

Message: 15
Date: Wed, 14 May 2008 18:57:17 -0400
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] OCT vs. isopentane
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4EBFF65383B74D49995298C4976D1D5E273D7B <@t> LSRIEXCH1.lsmaster.lifespan.org>

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

Isopentane is not an embedding medium. OCT (or another comparable brand) is the embedding medium. Isopentane cooled with liquid nitrogen or dry ice is the freezing medium for freezing the liquid OCT into solid blocks.

In sucrose cryoprotection, some people wash out the formalin before going into the first sucrose solution.  I find that the sucrose solution itself washes out the formalin quite adequately.  You do not wash at all between the different concentrations of sucrose.  That would defeat the purpose of using an ascending series of concentrations.

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

Message: 16
Date: Wed, 14 May 2008 17:31:20 -0600
From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
Subject: Re: [Histonet] Standardized Microtomes
To: "Amos Brooks" <amosbrooks <@t> gmail.com>,	"Rene J Buesa"
	<rjbuesa <@t> yahoo.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <006801c8b61a$9da832b0$6501a8c0 <@t> DHXTS541>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=original

When recutting a block or having one where the embedded tissue is a bit out whack for decent cutting alignment, then the x/y axis IS adjusted.  The blade angle is generally not adjusted,  only the x/y axis is changed.  Even moving a blade sideways to access a sharp, new edge can alter the x/y orientation slightly.  When this happens, reapproaching the blade, x/y adjustement may be needed to get first sections coming off the block - often the sections one absolutely must have.

We don't change the blade angle unless someone tweaks the lever (generally an inexperienced tech or student who has no clue what is going on but they love flipping levers around).  Sometimes a blade angle is readjusted from a new blade lot even though the manufacturer of the blade is the same.  The blade manufacturing process can introduce minute changes in blades.  When changing from high to low profile blades we readjust blade angle slightly at times - the low profiles being thinner in metal thickness and narrower width from top to bottom than the high profile.

Rembedding and refacing a block is not ideal. As Amos points out, the danger of losing the region of interest in the embedded tissue is just too high 
especially on a recut.   One can always turn a block in the holder, but that 
means some clever, careful x/y axis adjustment - we do this all the time.

 I vote for Amos's way as he just described it, and that is how we do it in our lab. We meaning "I" do it, the x/y axis way.  The lever tweakers are generally gently repirmanded and taught the correct way for where a blade angle works best for the preferred blade in the lab.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozemant MT

----- Original Message -----
From: "Amos Brooks" <amosbrooks <@t> gmail.com>
To: "Rene J Buesa" <rjbuesa <@t> yahoo.com>
Cc: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, May 14, 2008 4:42 PM
Subject: Re: [Histonet] Standardized Microtomes

Sorry Rene,
     I absolutly disagree with that. The exact opposite: *Not* modifying the
angle of your block holder to suit the block, is not good practice. You are
bound to loose tissue when cutting archived blocks, or blocks from other
labs or even slight variations within your own lab. If you don't adapt to
the blockyou are cutting. Without modifying the angle of the holder, one has
2 choices: Just cut thru it (refacing) or re-embedding and thereby needing
to re-trim. Both scenerios will loose tissue.
     Often I have cut a freshly embedded block where the cassette didn't sit
perfectly flat on the mold, otherwise the embedding was perfect. (Often due
to poorly dissected tissue with high points). This results in an angle that
is slightly askew. Adjusting the block holder is much faster than
re-embedding the block. If it is done properly you can actually modify the
block angle without ever cutting any tissue even on a previously cut block.

We're going to have to disagree on this,
Amos

On Wed, May 14, 2008 at 9:14 AM, Rene J Buesa <rjbuesa <@t> yahoo.com> wrote:

> All in all the practice of "changing the angle to match the block you have
> to recut as needed" is not a good practice.
> During it you could end loosing tissue when trying to "build" a new flat
> surface from a block that was originally cut at a different angle, while
> "sculping" the new surface.
> The best practice is to have all the microtomes cutting at the same angle
> and try to convince those HTs that say that "I have always cut with this
> angle" that they are flat wrong and a general angle can be good for the
> majority of blocks.
> René J.
>
_______________________________________________
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------------------------------

Message: 17
Date: Wed, 14 May 2008 20:02:15 -0400
From: "jstaruk" <jstaruk <@t> masshistology.com>
Subject: [Histonet] Help identifying LS174T tumor cells
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <FCAED7CE64894D43BB47EB008CBE960E <@t> JimPC>
Content-Type: text/plain;	charset="US-ASCII"

OK, I've been working on this for over a week and finally decided I need
help.  LS174T tumor cells (a human colon cancer cell line) were implanted
into mouse lymph nodes.  I'm trying to immuno-stain these implanted cells.
I tried a pan cytokeratin antibody and a CK20 cytokeratin antibody and
neither worked.  Has anyone here ever worked with this cell line?  Any
suggestions on how to differentiate these cells from the normal mouse cells?

Thanks!

Jim

_____________________
    Jim Staruk
Mass Histology Service
www.masshistology.com

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

Message: 18
Date: Wed, 14 May 2008 18:09:26 -0700
From: Gamal Akabani <gamal.akabani <@t> gmail.com>
Subject: [Histonet] Gamal Akabani has sent you a hi5 Friend Request
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <1478199894.95.1210813766874.JavaMail.root <@t> sfapp294>
Content-Type: text/plain; charset="UTF-8"

Gamal Akabani would like to be your friend on hi5!

   I set up a hi5 profile and I want to add you as a friend so we can
   share pictures and start building our network. First you need to join
   hi5! Once you join, you will have a chance to create a profile, share
   pictures, and find friends.
   Thanks,
   Gamal

                               [1]Join hi5!»

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

Message: 19
Date: Thu, 15 May 2008 01:46:09 +0000
From: koellingr <@t> comcast.net
Subject: Re: [Histonet] Help identifying LS174T tumor cells
To: "jstaruk" <jstaruk <@t> masshistology.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<051520080146.9807.482B95E1000D52F40000264F22134843739D09020704040A0105 <@t> comcast.net>

Content-Type: text/plain

Jim,
Haven't worked on that cell line in lymph nodes but in other parts of mice, xenografts and monolayers.  I'm assuming these are nude mice so the cells aren't simply being destroyed by host (mouse) immune response.  I'd stay away from cytokeratins.  CK20 can be downregulated in that line and there are cytokeratins that endogenously mark in lymph nodes.  That line I believe is presumed to be goblet cell associated.  I've looked at it with CEA and villin, things other than CK20 or pan cytokeratin that might traditionally mark colon.  Might try cdx2 besides CEA or villin.  I believe in ATTC that line is known as a large producer of CEA and state of the art (back when I did this) was to localize  LS174T's was radiolabelled CEA.  Never tried but MUC antibody or even a PAS or mucin histology stain might do.

Raymond Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "jstaruk" <jstaruk <@t> masshistology.com> 

> OK, I've been working on this for over a week and finally decided I need 
> help. LS174T tumor cells (a human colon cancer cell line) were implanted 
> into mouse lymph nodes. I'm trying to immuno-stain these implanted cells. 
> I tried a pan cytokeratin antibody and a CK20 cytokeratin antibody and 
> neither worked. Has anyone here ever worked with this cell line? Any 
> suggestions on how to differentiate these cells from the normal mouse cells? 
> 
> Thanks! 
> 
> Jim 
> 
> _____________________ 
> Jim Staruk 
> Mass Histology Service 
> www.masshistology.com 
> 
> _______________________________________________ 
> Histonet mailing list 
> Histonet <@t> lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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

Message: 20
Date: Thu, 15 May 2008 12:44:08 +0930
From: "Jim Manavis" <jim.manavis <@t> imvs.sa.gov.au>
Subject: FW: [Histonet] Help identifying LS174T tumor cells
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001c01c8b639$bd8796b0$636c140a <@t> itp36533>
Content-Type: text/plain;	charset="US-ASCII"

Jim

I've gone through something like this a few years ago. We attempted to
differentiate between host mouse cells and introduced human cancer cells. We
used the human mitochondrial marker successfully on FFPE tissue to
differentiate between the two. Initially we were using the Chemicon antibody
(Cat No. MAB1273) but then struck a snag with it, where with a particular
set of batch numbers it didn't work. We then turned to the Abcam antibody
(Cat No. ab3298) and have been using this ever since

Cheers

Jim

Jim Manavis
Laboratory Manager
Hanson Institute
Centre for Neurological Diseases
IMVS, Adelaide, SA, 5000
Australia
Phone: 61-08-8222-3668 / 0401120697
FAX: 61-08-8222 3392
email: jim.manavis <@t> imvs.sa.gov.au
Disclaimer: Not this little black duck! 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
koellingr <@t> comcast.net
Sent: Thursday, 15 May 2008 11:16 AM
To: jstaruk; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Help identifying LS174T tumor cells

Jim,
Haven't worked on that cell line in lymph nodes but in other parts of mice,
xenografts and monolayers.  I'm assuming these are nude mice so the cells
aren't simply being destroyed by host (mouse) immune response.  I'd stay
away from cytokeratins.  CK20 can be downregulated in that line and there
are cytokeratins that endogenously mark in lymph nodes.  That line I believe
is presumed to be goblet cell associated.  I've looked at it with CEA and
villin, things other than CK20 or pan cytokeratin that might traditionally
mark colon.  Might try cdx2 besides CEA or villin.  I believe in ATTC that
line is known as a large producer of CEA and state of the art (back when I
did this) was to localize  LS174T's was radiolabelled CEA.  Never tried but
MUC antibody or even a PAS or mucin histology stain might do.

Raymond Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "jstaruk" <jstaruk <@t> masshistology.com> 

> OK, I've been working on this for over a week and finally decided I need 
> help. LS174T tumor cells (a human colon cancer cell line) were implanted 
> into mouse lymph nodes. I'm trying to immuno-stain these implanted cells. 
> I tried a pan cytokeratin antibody and a CK20 cytokeratin antibody and 
> neither worked. Has anyone here ever worked with this cell line? Any 
> suggestions on how to differentiate these cells from the normal mouse
cells? 
> 
> Thanks! 
> 
> Jim 
> 
> _____________________ 
> Jim Staruk 
> Mass Histology Service 
> www.masshistology.com 
> 
> _______________________________________________ 
> Histonet mailing list 
> Histonet <@t> lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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

Message: 21
Date: Thu, 15 May 2008 08:05:05 +0400
From: "Anne van Binsbergen" <annigyg <@t> gmail.com>
Subject: Re: [Histonet] Suggestions on Equipment??
To: "Rene J Buesa" <rjbuesa <@t> yahoo.com>
Cc: histonet <@t> lists.utsouthwestern.edu, MICHELLE SEAGLE
	<mrsseagle <@t> yahoo.com>
Message-ID:
	<f8332fbe0805142105p7cc0a96cw337af7f8f4aea99c <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

it seems like Rene and I were cut from a similar AP mould
Leica Cryotat - go for the new one with the on board decontamination
Sakura Embedder - its for left handers and right handers and the cold plate
is a separate unit - neat and compact
Sakura DRS2000 or the Prisma (with the attached coverslipper) - it has small
staining dishes (optional) which i just love
Sakura need to vamp up their Cryostat technology - then my whole lab will be
a showcase of their products
 - are you reading this guys -
LOL
Annie
2008/5/15 Rene J Buesa <rjbuesa <@t> yahoo.com>:

> Cryostat from Leica; H&E stainer and embedding center from Sakura, although
> it is very likely that the Leica people (that manufacture the 3 pieces of
> equipment) will give you a better price if you buy all from them. Otherwise
> follow the first choice (Leica + Sakura).
>  René J.
>
> MICHELLE SEAGLE <mrsseagle <@t> yahoo.com> wrote:
>
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>

-- 
Anne van Binsbergen (Hope)
Abu Dhabi
UAE

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

Message: 22
Date: Thu, 15 May 2008 05:43:21 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Standardized Microtomes
To: Amos Brooks <amosbrooks <@t> gmail.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <812281.7705.qm <@t> web65711.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Amos:
  You had pointed out to the "acceptable exception". I do not advocate to NEVER align a blade to a block when necessary. What I say that for new block to be cut ALL microtomes in the lab should use the same angle and allow, for instance, that any HT could cut a recur for any special procedure without having to adjust  the cutting angle to that specific block, because it was cut with an uniform angle.
  Archival blocks, or those received from another institution as a consult, of course that should be cut by adapting the cutting angle to the block, that is always better than reembedding the block.
  So, for NEW, everyday routine blocks: all microtomes with the same angle.
  For old or consult blocks: adapt the microtome to them.
  Besides, you don't have to be sorry to express your opinion!
  René J.

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

Message: 23
Date: Thu, 15 May 2008 09:00:32 -0400
From: "Whitaker, Bonnie" <Bonnie.Whitaker <@t> osumc.edu>
Subject: [Histonet] Looking for Melissa Barrett
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <3CE20ED86C4A114EBDF3BCE8DEFD8F60262493 <@t> msxc06.OSUMC.EDU>
Content-Type: text/plain; charset=us-ascii

Hi All,
We are trying to locate Melissa Barrett (at one time she was at the South
Bend Medical Foundation) regarding a revision and repackaging of an exercise
she did for TechSample a few years back.  
Melissa, if you are here please email me with contact info, or if anyone else
knows how to reach her, please let me know.
Thanks,
Bonnie Whitaker
Histology Manager
Ohio State University Medical Center
614.293.5048

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

Message: 24
Date: Thu, 15 May 2008 10:33:45 -0400
From: "Rutledge, Nancy" <nrutledge <@t> CapeCodHealth.org>
Subject: [Histonet] need knife guard
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<47AD3B259E920D449F580E6AE82C2B8F210256 <@t> FHEXSVR2.FHDOMAIN1.capecodhealth.org>

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

I'm looking for one of the old knife guards, metal clip, fits over blade
that extends beyond knife holder.  Happy to pay for it, only need one.

Thanks

Nancy Rutledge

Falmouth Hospital

100 Ter Heun Drive

Falmouth,

MA 02540

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

Message: 25
Date: Thu, 15 May 2008 09:59:48 -0500
From: "Dawson, Glen" <GDawson <@t> dynacaremilwaukee.com>
Subject: RE: [Histonet] Salary Scales
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<B3D65550856D0146B900D401EE313D4B016C2CAF <@t> dynams.dynacaremilwaukee.com>

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

$26.20 / hour is "decent"?  For the Milwaukee area, that is a phenominal payrate.  What regions see this as "run of the mill"?  

I wish I could offer techs this pay.  It would certainly solve all of our staffing issues in one fell swoop.

I also wonder where payscales like this are when the Histology pay scales are being compiled for publication.  You look at our printed ranges and you cannot help but think that they are more useful to those who want to keep histo-pay low.

Call me a coward but I don't feel comfortable quoting our average histotech pay.  I'll just say that it is SIGNIFICANTLY below $26.20.

Glen Dawson  BS, HT & QIHC (ASCP)
IHC Manager
Milwaukee, WI

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Rene J
Buesa
Sent: Wednesday, May 14, 2008 2:22 PM
To: JR R; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Salary Scales

Which is equivalent to $26.20 / hour (2080 hours / year) and could be considered as "decent".
  René J.

JR R <rosenfeldtek <@t> hotmail.com> wrote:

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

Message: 26
Date: Thu, 15 May 2008 09:18:00 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: RE: [Histonet] Salary Scales
To: "Dawson, Glen" <GDawson <@t> dynacaremilwaukee.com>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <355672.50007.qm <@t> web65701.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Glen:
  And it will keep that way until histotechs star demanding what is deserved!

  Have you realized that histotechs are the only specialists in the medical lab that have to make decisions all along the process?
  When to reject a too thick slice of tissue to assure proper processing?
  What part to embed to cut?
  Up to where trim the block discarding parts of the specimen FOR EVER?!
  Which section to take or which to discard FOR EVER?!
  When to stop differentiation in a special stain?

  There is no other area of the ML that has to take so many decisions, and they are better paid. And will be until the HTs decide to take action and demand what is deserved.
  Just my opinion (as usual!).
  René J.

"Dawson, Glen" <GDawson <@t> dynacaremilwaukee.com> wrote:

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

Message: 27
Date: Thu, 15 May 2008 17:39:02 +0100
From: "Rupert Street" <rhdes.street <@t> googlemail.com>
Subject: [Histonet] Alternative Preserving Medium for Histology
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<2e02b98e0805150939y2eca73dcld8413d92c742e9d6 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Dear All;

At the risk of running the gamut with your news group, and I have asked
permission. Myself and my colleagues are looking for people interested in
beta testing an alternative to formalin/formaldehyde for within the
histopathology industry.

Before you start cracking back with questions, this product is not a
fixative so therefore is no use at all for long term preservation. It is
purely focussed towards biopsy usage.

The product has been adapted from a successful product used within the
funeral industry which seeks to replace formalin based embalming wherever
people have concerns etc.

If you are interested in helping us please send your details to the attached
e-mail.

Regards

Rupert Street

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

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End of Histonet Digest, Vol 54, Issue 21
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