Re: Fluorescent double label
Melissa
The thickness of your section really depends on what you need to see and,
if you are making images, the depth of field of the lenses you're using.
For example, if you're wanting to follow processes, you want thicker
sections. If you want sharp images at mags above 20x over as large an
area as possible, you'd want to cut your sections thinner.
If your primary antibodies are from different species, then apply them as
a cocktail and the secondary antibodies as well.
Fading of signal depends on the fluor you're using, how long your sections
are exposed to light, especially on the microscope, and how you store the
sections.
I've switched over to alexa fluors and am a happy camper (me and those
folks down in Pago Pago). They don't bleach as rapidly and at least the
488 is very bright. After you coverslip the slides, paint the edges with
finger nail polish. Store the slides at -20 deg C and you'll be a happy
camper, too.
John Carroll Dennis
Anatomy, Physiology, and Pharmacology
109 Greene Hall
Auburn University, AL 36849
On Fri, 27 Dec 2002, Melissa Gonzalez wrote:
> Hello,
> I have a few questions concerning fluorescent procedures on frozen tissue...
> 1. Do people recommend cutting tissue thicker for better visualization (10 microns instead of 5?)
> 2. For double labeling, should both antibodies be applied at once or separately? Does it make a difference?
> 3. How do you keep the signal from fading after one day?
>
> I am specifically trying to label CD4 and CD8 so if anyone has any previous experience with products and procedures that work I would really appreciate it.
>
> Thanks again
> Melissa
>
> -----Original Message-----
> From: HistoNet Server [mailto:histonet@pathology.swmed.edu]
> Sent: Thursday, December 26, 2002 9:58 PM
> To: HistoNet Server
> Subject: Daily Digest
>
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 02:47:18 -0600
> From: nyilmaz@mersin.edu.tr
> Subject: Acrilic Resin
>
> Hi Histonetters...
> We are planning to work with acrilic resin for immunoelectron microscopy. Is
> there anybody to help us about technical equipment and preparation protocol
> for acrilic resin? And if you give the web addresses related with acrilic
> resin procedure we will be so happy.
> Merry christmas.
>
> ___________________________________
> Mersin #220#niversitesi, http://www.mersin.edu.tr
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 08:03:02 -0600
> From: JHoffpa464@aol.com
> Subject: Re: The Reason for the Season
>
> heres wishing everyone a happy safe and prosperouse new years. for all those
> who over endulge, i have a sure fire hang over cure.
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 12:55:02 -0600
> From: Melissa Gonzalez
> Subject: Factor 8
>
> Hello, happy holidays everyone!
> I am still looking for a reliable source (and protocol if possible) for
> staining for Factor 8 expression in RAT hepatocytes. (We are looking for
> cytoplasmic staining, not endothelial) If anyone could help me out with this I
> would greatly appreciate it!!
>
> Thanks,
> Melissa
>
> - -----Original Message-----
> From: HistoNet Server [mailto:histonet@pathology.swmed.edu]
> Sent: Tuesday, December 24, 2002 9:57 PM
> To: HistoNet Server
> Subject: Daily Digest
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 00:08:19 -0600
> From: "Brothers, Julie {CNS~Palo Alto}"
> Subject: Out of Office AutoReply: Daily Digest
>
> Hello - I am currently out of the office and will be back on Monday, January
> 6th. I will not be checking my email.
>
> Thanks, Julie
>
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 02:59:06 -0600
> From: ALLISON@CARDIFF.AC.UK
> Subject: Individual characteristics
>
> Tolerance is a virtue
> Russ Allison,
> Dental School
> Cardiff
> Wales
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 07:25:50 -0600
> From: Vicki Gauch
> Subject: Happy Holidays
>
> I just wanted to wish everyone a very Merry Christmas, Happy Holidays, and a
> Happy, Healthy New Year !!! Thank you for all of your help over the past
> year...Hope you all know how much you are appreciated !!!
>
> Vicki Gauch
> AMCH
> Albany, NY
>
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 08:42:06 -0600
> From: AliNeumann@aol.com
> Subject: Variability of histology staining-request for input
>
>
> Hi everyone, I am the director of a new small outpatient laboratory, and we
> have had a very persistent and recalcitrant problem with variability of
> staining (some sections light, others dark, especially on eosin but to a
> lesser degree hematoxylin). We are soaking slides in xylene for 3 minutes in
> each of 3 bins with agitation, then staining them on a Varistain 24-4 with a
> regressive stain. Our staining protocols have been:
>
> 1. 100% Alcohol 20 seconds
> 2. 100% Alcohol 10 seconds
> 3. 95% Alcohol 10 seconds
> 4. Tap Water 10 seconds
> 5. Hematoxylin 7 minutes
> (Protocol Harris mercury free non-acidified type)
> 6. Tap Water 20 seconds
> 7. Acid Alcohol 2 seconds
> 8. Tap Water 20 seconds
> 9. Ammonia Water 2 minutes
> 10. Tap Water 10 seconds
> 11. 95% Alcohol 10 seconds
> 12. Eosin 15 seconds
> 13. 95% Alcohol 10 seconds
> 14. 95% Alcohol 10 seconds
> 15. 100% Alcohol 20 seconds
> 16. 100% Alcohol 20 seconds
>
>
> 1. 100% Ethanol 40 seconds
> 2. 100% Ethanol 40 seconds
> 3. 95% Ethanol 40 seconds
> 4. Tap water 60 seconds
> 5. Hematoxylin 10 minutes
> ( Protocol Harris mercury free non-acidified type)
> 6. Tap water (sink) 60 seconds
> 7. Acid Alcohol 2 seconds
> 8. Tap water (sink) 60 seconds
> 9. Ammonia water 2.5 minutes
> 10. Tap water 5 minutes
> 11. 95% EtOH 1 minute
> 12. Eosin 10 seconds
> 13. 95% EtOH 10 seconds
> 14. 95% EtOH 10 seconds
> 15. 100% EtOH 40 seconds
> 16. 100% EtOH 40 seconds
>
> We have had the same problem with both protocols, and process mostly small
> skin biopsies.
>
> Our acid and ammonia solutions are as follows:
>
> 1% acid alcohol
> 2000 ml of 70% EtOH
> 5 ml of HCl (hydrochloric acid)
>
> 1% Ammonia water
> 1980 cc tap water
> 40 ml of ammonium hydroxide
>
> We have tried longer xylene pre-treatment with more agitation, and wiping off
> alcohol drops before removing the bin from the last alcohol, as well as
> various staining adjustments. We process most of our tissue on a short 3
> hour cycle however believe this problem has persisted even on
> overnight-processed tissue.
> I would very much appreciate any advise any of you can give me on this
> problem. Thanks very much and have a happy holiday! Alice Neumann M.D.
> Precision Pathology PC, Arvada CO alineumann@aol.com 303-432-7855
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - - ------------------ MIME Information follows ------------------
>
>
> - - --Boundary_(ID_bhUjtUw8uDbvuUrCeMTk9Q)
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> LANG="0">Hi everyone, I am the director of a new small outpatient laboratory,
> and we have had a very persistent and recalcitrant problem with variability of
> staining (some sections light, others dark, especially on eosin but to a
> lesser degree hematoxylin). We are soaking slides in xylene for 3
> minutes in each of 3 bins with agitation, then staining them on a Varistain
> 24-4 with a regressive stain. Our staining protocols have been:
>
> 1. 100% Alcohol 20 seconds
> 2. 100% Alcohol 10 seconds
> 3. 95% Alcohol 10 seconds
> 4. Tap Water 10 seconds
> 5. Hematoxylin 7 minutes
> (Protocol Harris mercury free non-acidified
> type)
> 6. Tap Water 20 seconds
> 7. Acid Alcohol 2 seconds
> 8. Tap Water 20 seconds
> 9. Ammonia Water 2 minutes
> 10. Tap Water 10 seconds
> 11. 95% Alcohol 10 seconds
> 12. Eosin
> 15
> seconds
> 13. 95% Alcohol 10 seconds
> 14. 95% Alcohol 10 seconds
> 15. 100% Alcohol 20 seconds
> 16. 100% Alcohol 20 seconds
>
>
> 1. 100% Ethanol 40 seconds
> 2. 100% Ethanol 40 seconds
> 3. 95% Ethanol 40 seconds
> 4. Tap water
>
> 60 seconds
> 5. Hematoxylin 10 minutes
> ( Protocol Harris mercury free
> non-acidified type)
> 6. Tap water (sink) 60 seconds
> 7. Acid Alcohol 2 seconds
> 8. Tap water (sink) 60 seconds
> 9. Ammonia water 2.5 minutes
> 10. Tap water 5 minutes
> 11. 95% EtOH 1 minute
> 12. Eosin 10 seconds
> 13. 95% EtOH 10 seconds
> 14. 95% EtOH 10 seconds
> 15. 100% EtOH 40 seconds
> 16. 100% EtOH 40 seconds
>
> We have had the same problem with both protocols, and process mostly small
> skin biopsies.
>
> Our acid and ammonia solutions are as follows:
>
> 1% acid alcohol
> 2000 ml of 70% EtOH
> 5 ml of HCl (hydrochloric acid)
>
> 1% Ammonia water
> 1980 cc tap water
> 40 ml of ammonium hydroxide
>
> We have tried longer xylene pre-treatment with more agitation, and wiping off
> alcohol drops before removing the bin from the last alcohol, as well as
> various staining adjustments. We process most of our tissue on a short 3
> hour cycle however believe this problem has persisted even on
> overnight-processed tissue.
> I would very much appreciate any advise any of you can give me on this
> problem. Thanks very much and have a happy holiday! Alice
> Neumann M.D. Precision Pathology PC, Arvada CO alineumann@aol.com
> 303-432-7855
>
> - - --Boundary_(ID_bhUjtUw8uDbvuUrCeMTk9Q)--
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 08:56:25 -0600
> From: Kim Merriam
> Subject: Re: Variability of histology staining-request for input
>
>
> I would try longer rinses in water after the hematoxylin,
> acid alcohol and ammonia water (running water would probably
> be best). Also I would go longer in the 95% alcohols that
> are after the eosin.
>
> Kim Merriam
> TKT
> Cambridge, MA
>
>
>
> AliNeumann@aol.com wrote:
>
> > Hi everyone, I am the director of a new small outpatient
> > laboratory, and we have had a very persistent and
> > recalcitrant problem with variability of staining (some
> > sections light, others dark, especially on eosin but to a
> > lesser degree hematoxylin). We are soaking slides in
> > xylene for 3 minutes in each of 3 bins with agitation,
> > then staining them on a Varistain 24-4 with a regressive
> > stain. Our staining protocols have been:
> >
> > 1. 100% Alcohol 20 seconds
> > 2. 100% Alcohol 10 seconds
> > 3. 95% Alcohol 10 seconds
> > 4. Tap Water 10 seconds
> > 5. Hematoxylin 7 minutes
> > (Protocol Harris mercury free non-acidified type)
> > 6. Tap Water 20 seconds
> > 7. Acid Alcohol 2 seconds
> > 8. Tap Water 20 seconds
> > 9. Ammonia Water 2 minutes
> > 10. Tap Water 10 seconds
> > 11. 95% Alcohol 10 seconds
> > 12. Eosin 15 seconds
> > 13. 95% Alcohol 10 seconds
> > 14. 95% Alcohol 10 seconds
> > 15. 100% Alcohol 20 seconds
> > 16. 100% Alcohol 20 seconds
> >
> >
> > 1. 100% Ethanol 40 seconds
> > 2. 100% Ethanol 40 seconds
> > 3. 95% Ethanol 40 seconds
> > 4. Tap water 60 seconds
> > 5. Hematoxylin 10 minutes
> > ( Protocol Harris mercury free non-acidified type)
> > 6. Tap water (sink) 60 seconds
> > 7. Acid Alcohol 2 seconds
> > 8. Tap water (sink) 60 seconds
> > 9. Ammonia water 2.5 minutes
> > 10. Tap water 5 minutes
> > 11. 95% EtOH 1 minute
> > 12. Eosin 10 seconds
> > 13. 95% EtOH 10 seconds
> > 14. 95% EtOH 10 seconds
> > 15. 100% EtOH 40 seconds
> > 16. 100% EtOH 40 seconds
> >
> > We have had the same problem with both protocols, and
> > process mostly small skin biopsies.
> >
> > Our acid and ammonia solutions are as follows:
> >
> > 1% acid alcohol
> > 2000 ml of 70% EtOH
> > 5 ml of HCl (hydrochloric acid)
> >
> > 1% Ammonia water
> > 1980 cc tap water
> > 40 ml of ammonium hydroxide
> >
> > We have tried longer xylene pre-treatment with more
> > agitation, and wiping off alcohol drops before removing
> > the bin from the last alcohol, as well as various staining
> > adjustments. We process most of our tissue on a short 3
> > hour cycle however believe this problem has persisted even
> > on overnight-processed tissue.
> > I would very much appreciate any advise any of you can
> > give me on this problem. Thanks very much and have a
> > happy holiday! Alice Neumann M.D. Precision Pathology PC,
> > Arvada CO alineumann@aol.com 303-432-7855
>
>
>
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - - ------------------ MIME Information follows ------------------
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> <<<<<< See above "Message Body" >>>>>>
>
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>
> begin:vcard
> n:Merriam;Kim
> tel;fax:617-613-4012
> tel;work:617-503-0374
> x-mozilla-html:FALSE
> org:Transkaryotic Therapies;Animal Physiology
> adr:;;195 Albany St;Cambridge;MA;02139;
> version:2.1
> email;internet:kmerriam@tktx.com
> title:Research Specialist IV, Histology
> x-mozilla-cpt:;3
> fn:Kim Merriam, M.A., HT(ASCP)
> end:vcard
>
> - - --Boundary_(ID_x9OdrCGngQsHfHJ7Gj6amQ)--
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 09:32:44 -0600
> From: "Dawson, Glen"
> Subject: Christmas Eve Question
>
> Just out of curiosity, does anyone out there in histo-land still get
> Christmas Eve off as a holiday? We got it off up until 5 years ago when we
> became a reference lab. Now, all of our outside clients feel obliged to cut
> their staff loose for Christmas Eve and hammer my lab with all of their work
> with no feelings of guilt that I am working on a skeleton crew as well. I
> was lucky enough to put in an 11 hour day yesterday and have no idea when
> I'll get out of here today. I think that the Christmas spirit is something
> that reference lab histotechs can only hear about because we have no time to
> enjoy it ourselves. Ahhh, Christmas venting feels so good.
>
> Bah Humbug,
>
> Glen Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 09:44:02 -0600
> From: HACKERLAB@aol.com
> Subject: Happy Holidays
>
>
> Very Best Wishes for happy holidays, merry Christmas and a happy new year to
> everyone out there in histoland from all of us at hacker instruments &
> industries inc.
>
> may this time be peaceful and bright!
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - - ------------------ MIME Information follows ------------------
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>
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>
> <<<<<< See above "Message Body" >>>>>>
>
> - - --Boundary_(ID_cEnzMjvRYsdwRG03gELoNA)
> Content-type: text/html; charset=US-ASCII
> Content-transfer-encoding: 7BIT
>
> FACE="Balloon Bd BT" LANG="0">Very Best Wishes for happy holidays, merry
> Christmas and a happy new year to everyone out there in histoland from all of
> us at hacker instruments & industries inc.
>
> may this time be peaceful and bright!
>
> - - --Boundary_(ID_cEnzMjvRYsdwRG03gELoNA)--
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:06:29 -0600
> From: Kim Merriam
> Subject: [Fwd: Variability of histology staining-request for input]
>
>
>
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
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>
> Return-path:
> Delivery-receipt-to: Jacquie.Mack@CLS.ab.ca
> Received: from bimini.th.net (bimini.th.net [204.213.247.202])
> by mailbos1.tktx.com
> (iPlanet Messaging Server 5.2 HotFix 0.9 (built Jul 29 2002))
> with SMTP id <0H7M001PLRF7JG@mailbos1.tktx.com> for kmerriam@ims-ms-daemon
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> id ; Tue, 24 Dec 2002 08:42:33 -0700
> Date: Tue, 24 Dec 2002 08:42:32 -0700
> From: "Mack, Jacquie"
> Subject: RE: Variability of histology staining-request for input
> To: 'Kim Merriam'
> Message-id: <30C050525B881C4AAFF41E6D16543E6801762E99@mail3.mail.cls.ab.ca>
> MIME-version: 1.0
> X-Mailer: Internet Mail Service (5.5.2653.19)
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> Content-transfer-encoding: 7BIT
>
> I suspect that your alcohol times both going to water and coming back up to
> xylene are too short. Try increasing them to at least a minute or two each
> with agitation.
>
> Best of luck
> Jacquie
>
> - - -----Original Message-----
> From: Kim Merriam [mailto:kmerriam@tktx.com]
> Sent: December 24, 2002 7:53 AM
> To: AliNeumann@aol.com
> Cc: histonet@pathology.swmed.edu
> Subject: Re: Variability of histology staining-request for input
>
>
> I would try longer rinses in water after the hematoxylin,
> acid alcohol and ammonia water (running water would probably
> be best). Also I would go longer in the 95% alcohols that
> are after the eosin.
>
> Kim Merriam
> TKT
> Cambridge, MA
>
>
>
> AliNeumann@aol.com wrote:
>
> > Hi everyone, I am the director of a new small outpatient
> > laboratory, and we have had a very persistent and
> > recalcitrant problem with variability of staining (some
> > sections light, others dark, especially on eosin but to a
> > lesser degree hematoxylin). We are soaking slides in
> > xylene for 3 minutes in each of 3 bins with agitation,
> > then staining them on a Varistain 24-4 with a regressive
> > stain. Our staining protocols have been:
> >
> > 1. 100% Alcohol 20 seconds
> > 2. 100% Alcohol 10 seconds
> > 3. 95% Alcohol 10 seconds
> > 4. Tap Water 10 seconds
> > 5. Hematoxylin 7 minutes
> > (Protocol Harris mercury free non-acidified type)
> > 6. Tap Water 20 seconds
> > 7. Acid Alcohol 2 seconds
> > 8. Tap Water 20 seconds
> > 9. Ammonia Water 2 minutes
> > 10. Tap Water 10 seconds
> > 11. 95% Alcohol 10 seconds
> > 12. Eosin 15 seconds
> > 13. 95% Alcohol 10 seconds
> > 14. 95% Alcohol 10 seconds
> > 15. 100% Alcohol 20 seconds
> > 16. 100% Alcohol 20 seconds
> >
> >
> > 1. 100% Ethanol 40 seconds
> > 2. 100% Ethanol 40 seconds
> > 3. 95% Ethanol 40 seconds
> > 4. Tap water 60 seconds
> > 5. Hematoxylin 10 minutes
> > ( Protocol Harris mercury free non-acidified type)
> > 6. Tap water (sink) 60 seconds
> > 7. Acid Alcohol 2 seconds
> > 8. Tap water (sink) 60 seconds
> > 9. Ammonia water 2.5 minutes
> > 10. Tap water 5 minutes
> > 11. 95% EtOH 1 minute
> > 12. Eosin 10 seconds
> > 13. 95% EtOH 10 seconds
> > 14. 95% EtOH 10 seconds
> > 15. 100% EtOH 40 seconds
> > 16. 100% EtOH 40 seconds
> >
> > We have had the same problem with both protocols, and
> > process mostly small skin biopsies.
> >
> > Our acid and ammonia solutions are as follows:
> >
> > 1% acid alcohol
> > 2000 ml of 70% EtOH
> > 5 ml of HCl (hydrochloric acid)
> >
> > 1% Ammonia water
> > 1980 cc tap water
> > 40 ml of ammonium hydroxide
> >
> > We have tried longer xylene pre-treatment with more
> > agitation, and wiping off alcohol drops before removing
> > the bin from the last alcohol, as well as various staining
> > adjustments. We process most of our tissue on a short 3
> > hour cycle however believe this problem has persisted even
> > on overnight-processed tissue.
> > I would very much appreciate any advise any of you can
> > give me on this problem. Thanks very much and have a
> > happy holiday! Alice Neumann M.D. Precision Pathology PC,
> > Arvada CO alineumann@aol.com 303-432-7855
>
>
>
>
> - - --Boundary_(ID_rAfeoYsSdfgjOYiaLx7+Eg)
> Content-type: text/x-vcard; charset=us-ascii; name=kmerriam.vcf
> Content-transfer-encoding: 7BIT
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>
> begin:vcard
> n:Merriam;Kim
> tel;fax:617-613-4012
> tel;work:617-503-0374
> x-mozilla-html:FALSE
> org:Transkaryotic Therapies;Animal Physiology
> adr:;;195 Albany St;Cambridge;MA;02139;
> version:2.1
> email;internet:kmerriam@tktx.com
> title:Research Specialist IV, Histology
> x-mozilla-cpt:;3
> fn:Kim Merriam, M.A., HT(ASCP)
> end:vcard
>
> - - --Boundary_(ID_rAfeoYsSdfgjOYiaLx7+Eg)--
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:09:47 -0600
> From: "Morken, Tim"
> Subject: Half day on Chistmas eve.RE: Christmas Eve Question
>
> We get a half day off on Christmas eve, but only due to a Presidential
> directive.
>
> Tim Morken
> CDC, Atlanta
>
> - - -----Original Message-----
> From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com]
> Sent: Tuesday, December 24, 2002 10:21 AM
> To: histonet@pathology.swmed.edu
> Subject: Christmas Eve Question
>
>
> Just out of curiosity, does anyone out there in histo-land still get
> Christmas Eve off as a holiday? We got it off up until 5 years ago when we
> became a reference lab. Now, all of our outside clients feel obliged to cut
> their staff loose for Christmas Eve and hammer my lab with all of their work
> with no feelings of guilt that I am working on a skeleton crew as well. I
> was lucky enough to put in an 11 hour day yesterday and have no idea when
> I'll get out of here today. I think that the Christmas spirit is something
> that reference lab histotechs can only hear about because we have no time to
> enjoy it ourselves. Ahhh, Christmas venting feels so good.
>
> Bah Humbug,
>
> Glen Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:25:03 -0600
> From: Kari Bradshaw
> Subject: RE: Christmas Eve Question
>
> Christmas Eve is just like any other day here, except more people than usual
> are on vacation. Actually don't feel too bad...our lab runs with a
> pathologists and gross assistant on all Holidays that fall on a Monday;
> President's Day, Memorial Day, and Labor Day. Now that's tough!
>
> - - -----Original Message-----
> From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com]
> Sent: Tuesday, December 24, 2002 7:21 AM
> To: histonet@pathology.swmed.edu
> Subject: Christmas Eve Question
>
>
> Just out of curiosity, does anyone out there in histo-land still get
> Christmas Eve off as a holiday? We got it off up until 5 years ago when we
> became a reference lab. Now, all of our outside clients feel obliged to cut
> their staff loose for Christmas Eve and hammer my lab with all of their work
> with no feelings of guilt that I am working on a skeleton crew as well. I
> was lucky enough to put in an 11 hour day yesterday and have no idea when
> I'll get out of here today. I think that the Christmas spirit is something
> that reference lab histotechs can only hear about because we have no time to
> enjoy it ourselves. Ahhh, Christmas venting feels so good.
>
> Bah Humbug,
>
> Glen Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:25:21 -0600
> From: "Charles.Embrey"
> Subject: FW: The Reason for the Season
>
>
>
> - - -----Original Message-----
> From: Charles.Embrey
> Sent: Tuesday, December 24, 2002 8:24 AM
> To: 'sloanaroni@netscape.net'
> Subject: RE: The Reason for the Season
>
>
> A little cranky are we? Sorry but when I logged onto histonet I don't
> remember anywhere in the welcome e-mail that I was to strictly limit my
> correspondence to histology only. We are a community of people not
> histology robots. Use your delete key if the subject line doesn't interest
> you and quit your whining. smile a little and enjoy life.
> "You will be visited be three spirits, the first when the clock strikes
> one......."
> Charles R. Embrey Jr. PA(AAPA), HT(ASCP)
> Histology Manager
> Carle Clinic, Urbana Illinois
>
> - - -----Original Message-----
> From: sloanaroni@netscape.net [mailto:sloanaroni@netscape.net]
> Sent: Monday, December 23, 2002 3:12 PM
> To: CTague@ahs.llumc.edu; lesley@vancouverbc.net;
> bill501@mindspring.com; histonet@pathology.swmed.edu
> Subject: RE: The Reason for the Season
>
>
> How have we gone from Histology to Theology? This is not what this list
> server is for. I don't know about you guys but I don't have time for
> religious philisophical banter when I check my email-
>
>
>
> "Tague, Curtis" wrote:
>
> >Actually, the reason for the season is the BIRTH (not rebirth) of the SON
> (not sun).
> >
> >God bless you, my friend.
> >
> > -----Original Message-----
> >From: Lesley Weston [mailto:lesley@vancouverbc.net]
> >Sent: Sunday, December 22, 2002 09:24
> >To: Bill Blank; histonet@pathology.swmed.edu
> >Subject: Re: The Reason for the Season
> >
> >on 22/12/2002 8:43 AM, Bill Blank at bill501@mindspring.com wrote:
> >
> >>> I agree my sister. JESUS IS THE REASON FOR THE SEASON NOT JUST PRESENTS
> OR
> >>> SANTA CLAUS
> >>
> >> Actually the reason for the season is the rebirth of the Sun on the
> >> Winter Solstice.
> >>
> >
> >Mostly, the reason for the season is to cheer us all up because we've all
> >got SAD. This has tremendous relevance to histology.
> >
> >Lesley Weston.
> >
> >
> >
> >
> >
> >
>
>
> - - --
> "Great spirits have often encountered violent opposition from mediocre
> minds"
> A. Einstein
>
>
> __________________________________________________________________
> The NEW Netscape 7.0 browser is now available. Upgrade now!
> http://channels.netscape.com/ns/browsers/download.jsp
>
> Get your own FREE, personal Netscape Mail account today at
> http://webmail.netscape.com/
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:37:42 -0600
> From: JHoffpa464@aol.com
> Subject: Re: Half day on Chistmas eve.RE: Christmas Eve Question
>
> thats probly the only good thing the pres has done.
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:40:59 -0600
> From: "Steve Machin UK"
> Subject: Re: Christmas Eve Question
>
> I am in an English NHS lab and we don't get Christmas eve off.
> It's 4:30pm now; all the other staff left shortly after lunch so I
> am just "minding the ship&#untiltill 5:15.
> The place is very quiet although I understand that our out-patient
> clinics are likely to run until 6pm.
> So I will get two days off and then we will be back on Friday for
> work as usual.
> --- "Dawson, Glen" wrote: > Just out of
> curiosity, does anyone out there in histo-land still
> > get
> > Christmas Eve off as a holiday? We got it off up until 5 years ago
> > when we
> > became a reference lab. Now, all of our outside clients feel
> > obliged to cut
> > their staff loose for Christmas Eve and hammer my lab with all of
> > their work
> > with no feelings of guilt that I am working on a skeleton crew as
> > well. I
> > was lucky enough to put in an 11 hour day yesterday and have no
> > idea when
> > I'll get out of here today. I think that the Christmas spirit is
> > something
> > that reference lab histotechs can only hear about because we have
> > no time to
> > enjoy it ourselves. Ahhh, Christmas venting feels so good.
> >
> > Bah Humbug,
> >
> > Glen Dawson BS, HT & QIHC (ASCP)
> > Lead IHC Technologist
> > Milwaukee, WI
> >
>
> __________________________________________________
> Do You Yahoo!?
> Everything you'll ever need on one web page
> from News and Sport to Email and Music Charts
> http://uk.my.yahoo.com
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:47:24 -0600
> From: "Steve Machin UK"
> Subject: Merry Christmas?
>
> Merry Christmas to everyone.
> Lets hope its not the last one, which it could be if there is a big
> war in the Gulf.
>
> Steve Machin UK
>
> __________________________________________________
> Do You Yahoo!?
> Everything you'll ever need on one web page
> from News and Sport to Email and Music Charts
> http://uk.my.yahoo.com
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:50:36 -0600
> From: Instrumedics
> Subject: Re: Daily Digest
>
> To all our best wishes for a very good New Year!
>
> Bernice
> schiller@instrumedics.com
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 10:59:22 -0600
> From: "Weems, Joyce"
> Subject: RE: Christmas Eve Question
>
> We're here, a couple of staff short. Yesterday was a record workload day for
> us. It must be worse for the patients who we are working on - as we have
> some very seriously ill folk here.
>
> We have a lighter surgery day, but nevertheless full. We'll be back on
> Thurs.
>
> Blessings to all. j
>
>
> Joyce Weems
> Pathology Manager
> Saint Joseph's Hospital of Atlanta
> 404-851-7376
> 404-851-7831 - fax
>
> -----Original Message-----
> From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com]
> Sent: Tuesday, December 24, 2002 10:21 AM
> To: histonet@pathology.swmed.edu
> Subject: Christmas Eve Question
>
> Just out of curiosity, does anyone out there in histo-land still get
> Christmas Eve off as a holiday? We got it off up until 5 years ago when we
> became a reference lab. Now, all of our outside clients feel obliged to cut
> their staff loose for Christmas Eve and hammer my lab with all of their work
> with no feelings of guilt that I am working on a skeleton crew as well. I
> was lucky enough to put in an 11 hour day yesterday and have no idea when
> I'll get out of here today. I think that the Christmas spirit is something
> that reference lab histotechs can only hear about because we have no time to
> enjoy it ourselves. Ahhh, Christmas venting feels so good.
>
> Bah Humbug,
>
> Glen Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 11:15:44 -0600
> From: Kemp Watson
> Subject: RE: The Reason for the Season
>
> "WHAT ARE THE RULES?
> You may post any questions you wish pertaining to histology, pathology,
> in-situ hybridization, immunohistochemistry etc. Equipment and reagent
> evaluations, laboratory management issues, government regulations, and job
> opportunities are all appropriate topics. The University asks that we
> restrict
> the use of its hardware and software to business purposes only..."
>
> That was from my subscription. The reply from Curtis below may have been a
> bit terse indeed, but he's not wrong.
>
> I celebrate Christmas, but I'd hate to see the response from many on this
> list when it was deluged with Happy Hanukkah/Chanukah, Happy Diwali, Happy
> Ramadan, Happy Winter Solstice, Happy Eid al-Adha, etc. Non-Christians
> generally have a lot more awareness of religious diversity, it seems, at
> least regarding mailing list protocol.
>
> Happy New Year doesn't work, either...
>
> I'm not meaning to knock people's good wishes, mind you - I know they are
> well intended. Perhaps we could keep the good wishes to the first 3-4
> postings?
>
> Kemp Watson
>
> > -----Original Message-----
> > From: Charles.Embrey [mailto:Charles.Embrey@carle.com]
> > Sent: December 24, 2002 11:10 AM
> > To: 'histonet@pathology.swmed.edu'
> > Subject: FW: The Reason for the Season
> >
> >
> >
> >
> > -----Original Message-----
> > From: Charles.Embrey
> > Sent: Tuesday, December 24, 2002 8:24 AM
> > To: 'sloanaroni@netscape.net'
> > Subject: RE: The Reason for the Season
> >
> >
> > A little cranky are we? Sorry but when I logged onto histonet I don't
> > remember anywhere in the welcome e-mail that I was to strictly limit my
> > correspondence to histology only. We are a community of people not
> > histology robots. Use your delete key if the subject line
> > doesn't interest
> > you and quit your whining. smile a little and enjoy life.
> > "You will be visited be three spirits, the first when the clock strikes
> > one......."
> > Charles R. Embrey Jr. PA(AAPA), HT(ASCP)
> > Histology Manager
> > Carle Clinic, Urbana Illinois
> >
> > -----Original Message-----
> > From: sloanaroni@netscape.net [mailto:sloanaroni@netscape.net]
> > Sent: Monday, December 23, 2002 3:12 PM
> > To: CTague@ahs.llumc.edu; lesley@vancouverbc.net;
> > bill501@mindspring.com; histonet@pathology.swmed.edu
> > Subject: RE: The Reason for the Season
> >
> >
> > How have we gone from Histology to Theology? This is not what this list
> > server is for. I don't know about you guys but I don't have time for
> > religious philisophical banter when I check my email-
> >
> >
> >
> > "Tague, Curtis" wrote:
> >
> > >Actually, the reason for the season is the BIRTH (not rebirth) of the SON
> > (not sun).
> > >
> > >God bless you, my friend.
> > >
> > > -----Original Message-----
> > >From: Lesley Weston [mailto:lesley@vancouverbc.net]
> > >Sent: Sunday, December 22, 2002 09:24
> > >To: Bill Blank; histonet@pathology.swmed.edu
> > >Subject: Re: The Reason for the Season
> > >
> > >on 22/12/2002 8:43 AM, Bill Blank at bill501@mindspring.com wrote:
> > >
> > >>> I agree my sister. JESUS IS THE REASON FOR THE SEASON NOT
> > JUST PRESENTS
> > OR
> > >>> SANTA CLAUS
> > >>
> > >> Actually the reason for the season is the rebirth of the Sun on the
> > >> Winter Solstice.
> > >>
> > >
> > >Mostly, the reason for the season is to cheer us all up because we've all
> > >got SAD. This has tremendous relevance to histology.
> > >
> > >Lesley Weston.
> > >
> > >
> > >
> > >
> > >
> > >
> >
> >
> > --
> > "Great spirits have often encountered violent opposition from mediocre
> > minds"
> > A. Einstein
> >
> >
> > __________________________________________________________________
> > The NEW Netscape 7.0 browser is now available. Upgrade now!
> > http://channels.netscape.com/ns/browsers/download.jsp
> >
> > Get your own FREE, personal Netscape Mail account today at
> > http://webmail.netscape.com/
> >
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 11:51:01 -0600
> From: Bruce Gapinski
> Subject: RE: Christmas Eve Question
>
> Same here. I'm the supervisor and I sent everyone home by 9:00 AM. They
> went... kicking and screaming (You think I joke? No, I have the best
> staff)I'm doing glassware and waiting for the IHC to come out and I'm GONE.
> One World is Enough for All of Us!
> Bruce (I am not a rep) Gapinski HT(ASCP)
>
> - - -----Original Message-----
> From: Weems, Joyce [mailto:JWEEMS@sjha.org]
> Sent: Tuesday, December 24, 2002 8:48 AM
> To: 'Dawson, Glen'; histonet@pathology.swmed.edu
> Subject: RE: Christmas Eve Question
>
>
> We're here, a couple of staff short. Yesterday was a record workload day for
> us. It must be worse for the patients who we are working on - as we have
> some very seriously ill folk here.
>
> We have a lighter surgery day, but nevertheless full. We'll be back on
> Thurs.
>
> Blessings to all. j
>
>
> Joyce Weems
> Pathology Manager
> Saint Joseph's Hospital of Atlanta
> 404-851-7376
> 404-851-7831 - fax
>
> -----Original Message-----
> From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com]
> Sent: Tuesday, December 24, 2002 10:21 AM
> To: histonet@pathology.swmed.edu
> Subject: Christmas Eve Question
>
> Just out of curiosity, does anyone out there in histo-land still get
> Christmas Eve off as a holiday? We got it off up until 5 years ago when we
> became a reference lab. Now, all of our outside clients feel obliged to cut
> their staff loose for Christmas Eve and hammer my lab with all of their work
> with no feelings of guilt that I am working on a skeleton crew as well. I
> was lucky enough to put in an 11 hour day yesterday and have no idea when
> I'll get out of here today. I think that the Christmas spirit is something
> that reference lab histotechs can only hear about because we have no time to
> enjoy it ourselves. Ahhh, Christmas venting feels so good.
>
> Bah Humbug,
>
> Glen Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 11:58:23 -0600
> From: Valleygal@aol.com
> Subject: RE: The Reason for the Season
>
>
> You just have to love the personalities on this listserver!
> This is indeed a diverse group of folks - whatever your belief, be it
> religious, commercial, astrological or even if you are Scrooge...this is your
> time to celebrate and enjoy. So...let the celebration and enjoyment commence!
> Andi Grantham
> **********************************************************************
> =^..^=
>
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - - ------------------ MIME Information follows ------------------
>
>
> - - --Boundary_(ID_cHVE4uECYGEQ+6VIxduuwQ)
> Content-type: text/plain; charset=US-ASCII
> Content-transfer-encoding: 7BIT
>
> <<<<<< See above "Message Body" >>>>>>
>
> - - --Boundary_(ID_cHVE4uECYGEQ+6VIxduuwQ)
> Content-type: text/html; charset=US-ASCII
> Content-transfer-encoding: 7BIT
>
> FAMILY="SANSSERIF" FACE="Arial" LANG="0">
>
> - - --Boundary_(ID_cHVE4uECYGEQ+6VIxduuwQ)--
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 12:01:37 -0600
> From: Cynthia Favara
> Subject: RE: Individual characteristics
>
> Short, sweet and WISE!!!
>
> Peace to ALL
>
> Cynthia Favara
>
> - - -----Original Message-----
> From: RUSS ALLISON [mailto:Allison@Cardiff.ac.uk]
> Sent: Tuesday, December 24, 2002 1:51 AM
> To: histonet@pathology.swmed.edu
> Subject: Individual characteristics
>
>
> Tolerance is a virtue
> Russ Allison,
> Dental School
> Cardiff
> Wales
>
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 13:22:58 -0600
> From: "Dawson, Glen"
> Subject: RE: Christmas Eve
>
> All,
>
> Misery truly loves company. Thanx to all who let me know that I am not
> alone in my boat.
>
> Cheers,
>
> Glen Dawson
>
>
> - ----------------------------------------------------------------------
>
> Date: 24 Dec 2002 17:13:30 -0600
> From: Carlos Defeo
> Subject: Re:variability on staining
>
> Dear Ali:
> Can you tell the formula of the eosin solution you are using? For the
> changes in ethanol 95 I suppose you are employing an alcoholic solution.
> I recommend an aquous solution of eosin Y 0,75-1%
> and ommit then the wash in water,passing trough graded alcoholic baths. On
> this way you will have to change the first 95 ethanol dehydrating bath
> frequently but overdifferenciation of eosin will be avoided.
> Carlos,Montevideo,Uruguay.
>
>
> Here are the messages received yesterday!
>
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 13:41:38 -0600
> From: carl hobbs
> Subject: Variability of histology staining-request for input
>
>
>
> Tolerance is a virtue
> Russ Allison,
> Dental School
> Cardiff
> Wales
> So , when you get staining variability........hey! relax, no big deal. Be
> tolerant- as Russ says. Right, Russ? lol ....not quite the message you gave
> to your students a few years ago. As an Honourary member of The Musketeers,
> he was a Hard Man. ;-)
> Only kidding...it's Christmas.
> As already suggested, times are rather short. Relax, take longer ( at least
> double) in each step.....Dewax in a longer soak in xylene....ahhhhh, feels
> better already. Harris' is a HEAVY Hx, so take longer afterwards. Overstain
> in Eosin, then give the sections a relaxing, gentle dunk in each subsequent
> reagent ( no slacking, mind)
>
> Best wishes
>
>
> - ---
> Outgoing mail is certified Virus Free.
> Checked by AVG anti-virus system (http://www.grisoft.com).
> Version: 6.0.431 / Virus Database: 242 - Release Date: 17/12/02
>
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 14:48:57 -0600
> From: Mary Reeves
> Subject: Zinc Formalin and H&E Staining
>
> I have a pathologist who insists that fixing tissue in Zinc Formalin is
> causing poor nuclear detail and an overall cloudy appearance. Has anyone out
> there in histoland dealt with the issues caused by zinc formalin in there lab?
>
> Thanks for all the responses I am sure to receive!
>
> Mary Reeves
> Technical Specialist
> Histology
> 1-352-265-0680 ext 7-2113
>
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 15:33:12 -0600
> From: Bill Sinai
> Subject: Re: Zinc Formalin and H&E Staining
>
>
>
> Mary,
>
> It is more likely that the quality of the Formaldehyde is the problem rather
> then the zinc component. If you search the Histonet archives I am sure you
> will find many responses to this ongoing (world wide) problem.
> Bill Sinai
> Laboratory Manager
> Tissue Pathology
> ICPMR
> P.O. Box 533
> Wentworthville NSW 2145
> Ph 02 9845 7774
> > I have a pathologist who insists that fixing tissue in Zinc Formalin is
> causing poor nuclear detail and an overall cloudy appearance. Has anyone
> out there in histoland dealt with the issues caused by zinc formalin in
> there lab?
> > Thanks for all the responses I am sure to receive!
> >
> > Mary Reeves
> > Technical Specialist
> > Histology
> > 1-352-265-0680 ext 7-2113
> >
> >
>
>
> __________________________________________________________________
>
> This electronic message and any attachments may be confidential. If you
> are not the intended recipient of this message could you please delete the
> message and any attachments and advise the sender. Western Sydney
> Area Health Services (WSAHS) uses virus scanning software but excludes
> any liability for viruses contained in any email or attachment.
>
> This email may contain privileged and confidential information intended
> only for the use of the addressees named above. If you are not the
> intended recipient of this email, you are hereby notified that any use,
> dissemination, distribution, reproduction of this email is prohibited. If
> you have received this email in error, please notify WSAHS
> immediately.
>
> Any views expressed in this email are those of the individual sender
> except where the sender expressly and with authority states them
> to be the views of WSAHS.
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 17:52:38 -0600
> From: "Luck, Greg D."
> Subject: RE: slide and block labelers
>
> To all,
>
> I couldn't agree more on Dave's comment below. We have had our Surgipath
> cassette imprinter now for about 1 & 1/2 years. It has performed flawlessly
> thus far. It totally changed the gross room environment (ie. in a positive
> way). We used to discard approximately 10% of our cassettes and now I go
> days before I'll have to discard even one. No other single enhancement I
> have introduced into this department in the last 16 years as supervisor has
> been met with more unabashed and universal endorsement from histotechs,
> aides and pathologists alike. It's difficult for me to imagine how I could
> have spent that $7000 more effectively. Best wishes, Greg
>
> Greg Luck
> Anatomic Pathology Supervisor
> Deaconess Medical Center
> Spokane, WA 99204
> 509.473.7394
> luckg@empirehealth.org
>
>
> - -----Original Message-----
> From: Dave Low [mailto:lowman034@yahoo.com]
> Sent: Monday, December 16, 2002 5:55 PM
> To: Histology, Metro Lab; histonet posting address
> Subject: Re: slide and block labelers
>
>
> Hi Deb,
>
> We have Surgipath's slide and cassette printer. It
> only prints on block/slide at a time, but I like it.
> It is very simple and easy to use. Take care!
>
> Dave Low HT(ASCP)QIHC
> Elmendorf Medical Center/Path
>
> - --- "Histology, Metro Lab" wrote:
> > We are interested in getting the slide and block
> > labelers.
> > Does anyone out there use them and what do you think
> > of them? Please reply to all.
> > Thanks,
> > Deb
> > Metro Lab
> > Davenport, IA
> >
>
>
> __________________________________________________
> Do you Yahoo!?
> Yahoo! Mail Plus - Powerful. Affordable. Sign up now.
> http://mailplus.yahoo.com
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 18:31:53 -0600
> From: Bill
> Subject: Re: Zinc Formalin and H&E Staining
>
> At 3:40 PM -0500 12/26/02, Mary Reeves wrote:
> >I have a pathologist who insists that fixing tissue in Zinc Formalin
> >is causing poor nuclear detail and an overall cloudy appearance.
> >Has anyone out there in histoland dealt with the issues caused by
> >zinc formalin in there lab?
>
> Hmmmmm. We just switched to zinc formalin because we were getting the
> cloudy artifact and it has gone away completely. Nuclear detail has
> never been better and micro-chatter on colon polyps has disappeared.
>
> I am convinced that the phase of the moon during which a decision to
> make a change in a histology procedure is critical to the results
> achieved...
>
> BB
> - --
> ____________________
> Wm F Blank MD
> Heartland Laboratory, Inc
> Chaffee, MO 63740
>
>
> ----------------------------------------------------------------------
>
> Date: 26 Dec 2002 21:12:23 -0600
> From: Marysia33@aol.com
> Subject: Looking for Chris Venable
>
>
> Chris, your email doesn't go through to you. Please email me.
> Mary Mullinax
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - ------------------ MIME Information follows ------------------
>
>
> - --Boundary_(ID_HiNldVjQPh9t/PG78iTaug)
> Content-type: text/plain; charset=US-ASCII
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>
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> Content-transfer-encoding: 7BIT
>
> style="BACKGROUND-COLOR: #ffffff" SIZE=2 FAMILY="SANSSERIF" FACE="Arial"
> LANG="0">Chris, your email doesn't go through to you. Please email me.
> Mary Mullinax
>
> - --Boundary_(ID_HiNldVjQPh9t/PG78iTaug)--
>
>
> Here are the messages received yesterday!
>
>
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