[Histonet] RE: Histonet Digest, Vol 75, Issue 26

Sun Feb 21 19:19:04 CST 2010

 DRG's = Dorsal Root Ganglion... To clarify....healthcare is safe from these.

-----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: Saturday, February 20, 2010 1:01 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 75, Issue 26

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Today's Topics:

   1. IF staining on peritoneal macrophages (Lecorchick, William)
   2. EDTA (Webb, Dorothy L)
   3. Re: DRGs (Robert Richmond)
   4. Re: EDTA (Rene J Buesa)
   5. Tissue repository (Joao Pessoa)
   6. Re: Tissue repository (Rene J Buesa)
   7. seeking histology position (Pathrm35 <@t> comcast.net)
   8. Re: EDTA (gayle callis)
   9. RE: Re: EDTA (Rittman, Barry R)
  10. RE: Re: EDTA (gayle callis)
  11. Re: Distance learning? (Joao Pessoa)
  12. Re: Re: DRGs (John Kiernan)
  13. shandon's formal fixx and cytoblock kit (shehnaz khan)

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

Message: 1
Date: Fri, 19 Feb 2010 13:17:51 -0500
From: "Lecorchick, William" <wlecorch <@t> rwjuhh.edu>
Subject: [Histonet] IF staining on peritoneal macrophages
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<09411E0112A96A459D8D5FBDAB9C15C71A4AB78B43 <@t> HAMEXMBA.rwjham.local>
Content-Type: text/plain; charset="us-ascii"

You may want to try using the "NuView" Cyto prep technique from QC sciences it is a good alternative to a Cytospin

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

Message: 2
Date: Fri, 19 Feb 2010 12:29:01 -0600
From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] EDTA
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<65365F35C0F2EF4D846EC3CA73E49C43C57696D718 <@t> HPEMX3.HealthPartners.int>
Content-Type: text/plain; charset="us-ascii"

I am looking into the various decals on the market and have found one that in addition to formic acid, has EDTA in the mix.  I have never worked with EDTA so would appre ciate any help in your comments on the use of EDTA in decalcification methods for bone marrow and routine specimens.

Thank you ahead of time for your advice!

Dorothy Webb, HT (ASCP)
Regions Histology Technical Supervisor
651-254-2962

  ________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.

If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0

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

Message: 3
Date: Fri, 19 Feb 2010 14:07:37 -0500
From: Robert Richmond <rsrichmond <@t> gmail.com>
Subject: [Histonet] Re: DRGs
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<abea52a61002191107v5663557s1fa87396e2c538fd <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

DRG's for mice? Do mice have Medicare now? I hope the pending health care reform legislation addresses this issue - it's no wonder costs are spiraling out of sight!

I have the feeling that a mouse DRG might be something other than "Diagnosis Related Group", the coding scheme that has determined Medicare (and many other insurers') payments to hospitals for close to thirty years now.

Histonet includes many people from diverse disciplines. It's better not to assume that any but the commonest abbreviations (H & E and not too many more) will be understood by every Histonetter.

Bob Richmond
Samurai Pathologist
Knoxville TN (I mean Tennessee)

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

Message: 4
Date: Fri, 19 Feb 2010 13:03:27 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] EDTA
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>, 	Dorothy LWebb
	<Dorothy.L.Webb <@t> HealthPartners.Com>
Message-ID: <796184.91241.qm <@t> web65714.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

EDTA (which stands for Ethylene-diamino-tratacetic-acid) is a chelating agent and the method of choice to decalcify BM biopsies.
The thing is that it should be used alone, and not combined with any acid, as you state (not even formic acid).
By itself will produce an extremely gentle decalcification that will be completely suitable for IHC studies.
René J.

--- On Fri, 2/19/10, Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com> wrote:

From: Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] EDTA
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Date: Friday, February 19, 2010, 1:29 PM

I am looking into the various decals on the market and have found one that in addition to formic acid, has EDTA in the mix.  I have never worked with EDTA so would appre ciate any help in your comments on the use of EDTA in decalcification methods for bone marrow and routine specimens.

Thank you ahead of time for your advice!

Dorothy Webb, HT (ASCP)
Regions Histology Technical Supervisor
651-254-2962

  ________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.

If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 _______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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

Message: 5
Date: Fri, 19 Feb 2010 13:21:09 -0800
From: Joao Pessoa <jphistology <@t> gmail.com>
Subject: [Histonet] Tissue repository
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<9e4d12d1002191321o6a98e4f6o481f75b6c603c393 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Dear histonet,

Does anyone know where I can find a tissue repository to exchange tissue blocks?  For example, I have some extra melanoma FFPE tissue blocks but would like some normal FFPE tonsil tissue blocks.  I do not wish to buy blocks, but rather trade.  Please let me know if there is such a place.

Thanks,

Joao
Histo Tech

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

Message: 6
Date: Fri, 19 Feb 2010 13:25:07 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Tissue repository
To: histonet <@t> lists.utsouthwestern.edu, Joao Pessoa
	<jphistology <@t> gmail.com>
Message-ID: <464358.67154.qm <@t> web65702.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Contact the NSH that I think offers such a service.
René J.

--- On Fri, 2/19/10, Joao Pessoa <jphistology <@t> gmail.com> wrote:

From: Joao Pessoa <jphistology <@t> gmail.com>
Subject: [Histonet] Tissue repository
To: histonet <@t> lists.utsouthwestern.edu
Date: Friday, February 19, 2010, 4:21 PM

Dear histonet,

Does anyone know where I can find a tissue repository to exchange tissue blocks?  For example, I have some extra melanoma FFPE tissue blocks but would like some normal FFPE tonsil tissue blocks.  I do not wish to buy blocks, but rather trade.  Please let me know if there is such a place.

Thanks,

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

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

Message: 7
Date: Fri, 19 Feb 2010 22:03:50 +0000 (UTC)
From: Pathrm35 <@t> comcast.net
Subject: [Histonet] seeking histology position
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<114145463.6366871266617030451.JavaMail.root <@t> sz0123a.westchester.pa.mail.comcast.net>

Content-Type: text/plain; charset=utf-8

Fellow techs, 

I am currently seekingÂ a F/T position as a bench tech. I am ASCP certified and Florida licensed w/ a BS degree.Â Relocation and temp. positions are a possibility. Please feel free to pass this e mail along if you know of anyone looking for a tech. 

Thanks in advance 

Ron Martin 

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

Message: 8
Date: Fri, 19 Feb 2010 15:03:46 -0700
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] Re: EDTA
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000101cab1af$698cf8c0$3ca6ea40$@callis <@t> bresnan.net>
Content-Type: text/plain;	charset="us-ascii"

You wrote: 

I am looking into the various decals on the market and have found one that in addition to formic acid, has EDTA in the mix.  I have never worked with EDTA so would appre ciate any help in your comments on the use of EDTA in decalcification methods for bone marrow and routine specimens.

****************************************************************************
****************************************************************************
**

The purpose of EDTA in an acid decalcifier is probably NOT performing the actual removal of calcium (or very little)  from the bone but chelating calcium ionized from bone by the acid that then (the calcium) settles to the bottom of the container.  I doubt very much that you will be dealing with
EDTA but with formic acid decalcification.   People who have developed acid
decalcifying solutions, with either formic acid or hydrochloric acid tout that the bone section looks very good, better that if EDTA is not present in the solution.  I never tested this claim in our bone studies, but it might be something to try, an acid solution containing EDTA and one without EDTA (keeping the same kind of acid in the study).  

 The acid is going to ionize the calcium from bone at a faster rate than the EDTA can chelate the calcium.  But most importantly, EDTA does not work in a low pH environment.  The chemistry of how EDTA acts at different pHs is well documented by chemists, and found in book chapters on EDTA.  My physical chemist spouse supplied me with such a chapter.  The pH of formic acid is what, pH 3 or so, and if so, EDTA only begins to chelate calcium around pH 4, and when fully protonated at pH 8, decalcifies faster at that pH than at pH 4, or below pH7.  The working pH for most EDTA solutions is commonly 7 to 7.6, but when the pH becomes more alkaline ( and 8 is going into alkaline
range)  then alkaline sensitive protein linkages can be damaged.  

EDTA, correctly written out by Rene Buesa e.g. ethylene diaminatetraacetiic acid comes in several molecular weights depending on whether is it EDTA without attached sodium, EDTA disodium and EDTA tetrasodium.  Tetrasodium is very soluble in water or PBS, but has a very alkaline pH that requires adjustment down to pH 7 to 7.6 or that high pH will damage alkaline sensitive protein linkages.  EDTA and disodium EDTA are not as soluble, usually no more than a 10% solution, requiring heat or addition of sodium hydroxide in order to dissolve.  EDTA is also expensive,  not affected by heat up to 60C but only if the bone is totally fixed with NBF.  I would not advise using 60C during lengthy EDTA decalcification, and possible damage to heat labile antigens.  There are publications on using EDTA to decalcify fresh bone samples, then snap freeze, cut frozen sections, then acetone fix the sections for murine CD markers compromised by both NBF and acid decalcification.  

EDTA mixtures are found in Histonet Archives,  histotechnology books and on websites e.g. IHCworld, as a chelating agent, it is a very slow decalcifier.
More advantages of EDTA are - no damage to antigens, nucleic acid staining, nor other tissue components.  It does affect the enzyme histochemical stain for alkaline phosphatase, and magnesium ions (chelated by EDTA) must be replaced in the staining solution.  Most clinical laboratories prefer to use formic acid or hydrochloric acid decalcifying solutions instead of EDTA when rapid diagnosis is needed.  

If you try EDTA alone, rinse the bone well (after decalcification) with running tap water since residual, excess EDTA in tissue precipitates in presence of alcohol, and the ppt makes the tissue difficult to section.
Endpoint determinations for complete removal of calcium with EDTA cannot be done chemically, although there is a weight gain/weight loss method that we use routinely that works or use an Xray machine, Faxitron.  You can buy EDTA mixtures from Poly Scientific and some other suppliers, or make it up yourself.  

Otherwise, try the formic acid with EDTA, it may work very well for you while not sacrificing speed for diagnosis. Formic acid will be less damaging to antigens than hydrochloric acid, and is a popular decalcifier because it is gentler than HCl. 

Good luck

Gayle M. Callis

HTL/HT/MT(ASCP)

Bozeman MT 59715

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

Message: 9
Date: Fri, 19 Feb 2010 16:20:25 -0600
From: "Rittman, Barry R" <Barry.R.Rittman <@t> uth.tmc.edu>
Subject: RE: [Histonet] Re: EDTA
To: 'Histonet' <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<75A0543E23D3A7458012D9E02EDBEC00098CEE3112 <@t> UTHCMS1.uthouston.edu>
Content-Type: text/plain; charset="us-ascii"

I agree with Gayle and Rene
I would not even use a mixture of the two.
Formic acid demineralization will work fine, the theory of using EDTA with it makes no sense.
EDTA demineralization usually uses the disodium salt. EDTA by itself is not very soluble but its sodium salts are.
It can be present as mono-, di-, tri- or tetra-sodium salts depending on the pH. At pH around 11 there are 4 groups that can be replaced with for example calcium.
Usually the disodium salt of EDTA is used as the tri and tetrasodium salts are only present in very alkaline solutions that tend to cause tissue maceration.
At acidic pH the EDTA will only work slowly and have one group replaceable as the monosodium EDTA.
There are several good formulae for using EDTA and formic acid in separate formulae, depending on your specific needs..
Barry

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of gayle callis [gayle.callis <@t> bresnan.net]
Sent: Friday, February 19, 2010 4:03 PM
To: 'Histonet'
Subject: [Histonet] Re: EDTA

You wrote:

I am looking into the various decals on the market and have found one that in addition to formic acid, has EDTA in the mix.  I have never worked with EDTA so would appre ciate any help in your comments on the use of EDTA in decalcification methods for bone marrow and routine specimens.

****************************************************************************
****************************************************************************
**

The purpose of EDTA in an acid decalcifier is probably NOT performing the actual removal of calcium (or very little)  from the bone but chelating calcium ionized from bone by the acid that then (the calcium) settles to the bottom of the container.  I doubt very much that you will be dealing with
EDTA but with formic acid decalcification.   People who have developed acid
decalcifying solutions, with either formic acid or hydrochloric acid tout that the bone section looks very good, better that if EDTA is not present in the solution.  I never tested this claim in our bone studies, but it might be something to try, an acid solution containing EDTA and one without EDTA (keeping the same kind of acid in the study).

 The acid is going to ionize the calcium from bone at a faster rate than the EDTA can chelate the calcium.  But most importantly, EDTA does not work in a low pH environment.  The chemistry of how EDTA acts at different pHs is well documented by chemists, and found in book chapters on EDTA.  My physical chemist spouse supplied me with such a chapter.  The pH of formic acid is what, pH 3 or so, and if so, EDTA only begins to chelate calcium around pH 4, and when fully protonated at pH 8, decalcifies faster at that pH than at pH 4, or below pH7.  The working pH for most EDTA solutions is commonly 7 to 7.6, but when the pH becomes more alkaline ( and 8 is going into alkaline
range)  then alkaline sensitive protein linkages can be damaged.

EDTA, correctly written out by Rene Buesa e.g. ethylene diaminatetraacetiic acid comes in several molecular weights depending on whether is it EDTA without attached sodium, EDTA disodium and EDTA tetrasodium.  Tetrasodium is very soluble in water or PBS, but has a very alkaline pH that requires adjustment down to pH 7 to 7.6 or that high pH will damage alkaline sensitive protein linkages.  EDTA and disodium EDTA are not as soluble, usually no more than a 10% solution, requiring heat or addition of sodium hydroxide in order to dissolve.  EDTA is also expensive,  not affected by heat up to 60C but only if the bone is totally fixed with NBF.  I would not advise using 60C during lengthy EDTA decalcification, and possible damage to heat labile antigens.  There are publications on using EDTA to decalcify fresh bone samples, then snap freeze, cut frozen sections, then acetone fix the sections for murine CD markers compromised by both NBF and acid decalcification.

EDTA mixtures are found in Histonet Archives,  histotechnology books and on websites e.g. IHCworld, as a chelating agent, it is a very slow decalcifier.
More advantages of EDTA are - no damage to antigens, nucleic acid staining, nor other tissue components.  It does affect the enzyme histochemical stain for alkaline phosphatase, and magnesium ions (chelated by EDTA) must be replaced in the staining solution.  Most clinical laboratories prefer to use formic acid or hydrochloric acid decalcifying solutions instead of EDTA when rapid diagnosis is needed.

If you try EDTA alone, rinse the bone well (after decalcification) with running tap water since residual, excess EDTA in tissue precipitates in presence of alcohol, and the ppt makes the tissue difficult to section.
Endpoint determinations for complete removal of calcium with EDTA cannot be done chemically, although there is a weight gain/weight loss method that we use routinely that works or use an Xray machine, Faxitron.  You can buy EDTA mixtures from Poly Scientific and some other suppliers, or make it up yourself.

Otherwise, try the formic acid with EDTA, it may work very well for you while not sacrificing speed for diagnosis. Formic acid will be less damaging to antigens than hydrochloric acid, and is a popular decalcifier because it is gentler than HCl.

Good luck

Gayle M. Callis

HTL/HT/MT(ASCP)

Bozeman MT 59715

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

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

Message: 10
Date: Fri, 19 Feb 2010 16:32:13 -0700
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: RE: [Histonet] Re: EDTA
To: "'Rittman, Barry R'" <Barry.R.Rittman <@t> uth.tmc.edu>,	"'Histonet'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000301cab1bb$c4e91210$4ebb3630$@callis <@t> bresnan.net>
Content-Type: text/plain;	charset="us-ascii"

We use EDTA tetra-sodium salt, pH 7.6 but the pH is adjusted down with glacial acetic acid using continuous stirring and a pH electrode in the solution (titration with the acid). This is a recipe from a Dr. Webster
(Webb) S. S. Jee publication. 

We like this since this 14% EDTA dissolves immediately in water or PBS compared disodium EDTA, and at a higher concentration = more molecules of EDTA available.  

We are very careful to NOT use this 14% tetrasodium EDTA at it original pH of around 10 or more.  

Decalcification is still very slow, but a recent bone project required EDTA decalcification (so we thought!) to protect antigens never stained before in our lab.  After trying both the EDTA and 10% formic acid, we found the antigen to be very robust after formic acid decalcification too. Lesson learned, don't put off a pilot study to test IHC after acid decalcification.
The formic acid would have been faster and cheaper.    

Gayle Callis 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R
Sent: Friday, February 19, 2010 3:20 PM
To: 'Histonet'
Subject: RE: [Histonet] Re: EDTA

I agree with Gayle and Rene
I would not even use a mixture of the two.
Formic acid demineralization will work fine, the theory of using EDTA with it makes no sense.
EDTA demineralization usually uses the disodium salt. EDTA by itself is not very soluble but its sodium salts are.
It can be present as mono-, di-, tri- or tetra-sodium salts depending on the pH. At pH around 11 there are 4 groups that can be replaced with for example calcium.
Usually the disodium salt of EDTA is used as the tri and tetrasodium salts are only present in very alkaline solutions that tend to cause tissue maceration.
At acidic pH the EDTA will only work slowly and have one group replaceable as the monosodium EDTA.
There are several good formulae for using EDTA and formic acid in separate formulae, depending on your specific needs..
Barry

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu
[histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of gayle callis [gayle.callis <@t> bresnan.net]
Sent: Friday, February 19, 2010 4:03 PM
To: 'Histonet'
Subject: [Histonet] Re: EDTA

You wrote:

I am looking into the various decals on the market and have found one that in addition to formic acid, has EDTA in the mix.  I have never worked with EDTA so would appre ciate any help in your comments on the use of EDTA in decalcification methods for bone marrow and routine specimens.

****************************************************************************
****************************************************************************
**

The purpose of EDTA in an acid decalcifier is probably NOT performing the actual removal of calcium (or very little)  from the bone but chelating calcium ionized from bone by the acid that then (the calcium) settles to the bottom of the container.  I doubt very much that you will be dealing with
EDTA but with formic acid decalcification.   People who have developed acid
decalcifying solutions, with either formic acid or hydrochloric acid tout that the bone section looks very good, better that if EDTA is not present in the solution.  I never tested this claim in our bone studies, but it might be something to try, an acid solution containing EDTA and one without EDTA (keeping the same kind of acid in the study).

 The acid is going to ionize the calcium from bone at a faster rate than the EDTA can chelate the calcium.  But most importantly, EDTA does not work in a low pH environment.  The chemistry of how EDTA acts at different pHs is well documented by chemists, and found in book chapters on EDTA.  My physical chemist spouse supplied me with such a chapter.  The pH of formic acid is what, pH 3 or so, and if so, EDTA only begins to chelate calcium around pH 4, and when fully protonated at pH 8, decalcifies faster at that pH than at pH 4, or below pH7.  The working pH for most EDTA solutions is commonly 7 to 7.6, but when the pH becomes more alkaline ( and 8 is going into alkaline
range)  then alkaline sensitive protein linkages can be damaged.

EDTA, correctly written out by Rene Buesa e.g. ethylene diaminatetraacetiic acid comes in several molecular weights depending on whether is it EDTA without attached sodium, EDTA disodium and EDTA tetrasodium.  Tetrasodium is very soluble in water or PBS, but has a very alkaline pH that requires adjustment down to pH 7 to 7.6 or that high pH will damage alkaline sensitive protein linkages.  EDTA and disodium EDTA are not as soluble, usually no more than a 10% solution, requiring heat or addition of sodium hydroxide in order to dissolve.  EDTA is also expensive,  not affected by heat up to 60C but only if the bone is totally fixed with NBF.  I would not advise using 60C during lengthy EDTA decalcification, and possible damage to heat labile antigens.  There are publications on using EDTA to decalcify fresh bone samples, then snap freeze, cut frozen sections, then acetone fix the sections for murine CD markers compromised by both NBF and acid decalcification.

EDTA mixtures are found in Histonet Archives,  histotechnology books and on websites e.g. IHCworld, as a chelating agent, it is a very slow decalcifier.
More advantages of EDTA are - no damage to antigens, nucleic acid staining, nor other tissue components.  It does affect the enzyme histochemical stain for alkaline phosphatase, and magnesium ions (chelated by EDTA) must be replaced in the staining solution.  Most clinical laboratories prefer to use formic acid or hydrochloric acid decalcifying solutions instead of EDTA when rapid diagnosis is needed.

If you try EDTA alone, rinse the bone well (after decalcification) with running tap water since residual, excess EDTA in tissue precipitates in presence of alcohol, and the ppt makes the tissue difficult to section.
Endpoint determinations for complete removal of calcium with EDTA cannot be done chemically, although there is a weight gain/weight loss method that we use routinely that works or use an Xray machine, Faxitron.  You can buy EDTA mixtures from Poly Scientific and some other suppliers, or make it up yourself.

Otherwise, try the formic acid with EDTA, it may work very well for you while not sacrificing speed for diagnosis. Formic acid will be less damaging to antigens than hydrochloric acid, and is a popular decalcifier because it is gentler than HCl.

Good luck

Gayle M. Callis

HTL/HT/MT(ASCP)

Bozeman MT 59715

_______________________________________________
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

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

Message: 11
Date: Fri, 19 Feb 2010 20:46:28 -0800
From: Joao Pessoa <jphistology <@t> gmail.com>
Subject: Re: [Histonet] Distance learning?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<9e4d12d1002192046h29bc5cfbo48367b74d9a36bf9 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

You should try Harford Community College.  I know several histo techs who have completed histo tech training through the Harford on-line program, and the practical or technical portions in their own lab (employer must approve).

http://www.harford.edu/cet/histotech/default.asp

Thanks,

Joao
Histo Tech

On Fri, Feb 12, 2010 at 7:25 AM, Jay Lundgren <jaylundgren <@t> gmail.com> wrote:

>  A few years ago I remember hearing about a NAACLS approved, distance 
> learning Histotechnology program.  They were using the Internet and 
> teleconferencing to train students.  Is anyone still doing this, and 
> could you tell me where?
>
>                                                     Thanks for your 
> time,
>
>                                                                   Jay A.
> Lundgren, M.S., HTL (ASCP)
>  _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>

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

Message: 12
Date: Sat, 20 Feb 2010 02:40:54 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] Re: DRGs
To: Robert Richmond <rsrichmond <@t> gmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <fc12d9025ac4.4b7f4bb6 <@t> uwo.ca>
Content-Type: text/plain; CHARSET=US-ASCII

Good on yer, Bob! 
There are indeed far too many abbreviations. Anything  not in the Abbreviations appendix of an ordinary dictionary (volume = one litre or less, as on the average family's shelf) should be explained.

In this case my interpretation was DRG = dorsal root ganglia. Dissecting them out of rats is a difficult job even for the easiest one (C2), and a rat is 10 times as big as a mouse! The only easy-to-remove rat or mouse sensory ganglion is the trigeminal. 

Cheers,   John

John Kiernan
Anatomy, UWO
London, Canada
= = =
----- Original Message -----
From: Robert Richmond <rsrichmond <@t> gmail.com>
Date: Friday, February 19, 2010 14:08
Subject: [Histonet] Re: DRGs
To: histonet <@t> lists.utsouthwestern.edu

> DRG's for mice? Do mice have Medicare now? I hope the pending health 
> care reform legislation addresses this issue - it's no wonder costs 
> are spiraling out of sight!
> 
> I have the feeling that a mouse DRG might be something other than 
> "Diagnosis Related Group", the coding scheme that has determined 
> Medicare (and many other insurers') payments to hospitals for close to 
> thirty years now.
> 
> Histonet includes many people from diverse disciplines. It's better 
> not to assume that any but the commonest abbreviations (H & E and not 
> too many more) will be understood by every Histonetter.
> 
> Bob Richmond
> Samurai Pathologist
> Knoxville TN (I mean Tennessee)
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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

Message: 13
Date: Sat, 20 Feb 2010 15:54:32 +0200
From: shehnaz khan <shehnazster <@t> gmail.com>
Subject: [Histonet] shandon's formal fixx and cytoblock kit
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<4fe9f16e1002200554r1ff45e23q3a1ba567c24cb8e7 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi Histonetters

Could someone kindly share their views on the  shandon's formal fixx
(concentrate) and cytoblock kit for cell block preparation in cytology?  Has it been effective for cellular preservation and cell capture from inadeduate samples?

Thanking you in advance.

S .Khan
Dept of Cytology
University of Witwatersrand
Johannesburg
South Africa

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

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End of Histonet Digest, Vol 75, Issue 26
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