Fluorescent double label

From:Melissa Gonzalez

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



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contained in the following MIME Information.
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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 ------------------ This is a multi-part message in MIME format. - - --Boundary_(ID_x9OdrCGngQsHfHJ7Gj6amQ) Content-type: text/plain; charset=us-ascii; x-mac-type=54455854; x-mac-creator=4D4F5353 Content-transfer-encoding: 7BIT <<<<<< See above "Message Body" >>>>>> - - --Boundary_(ID_x9OdrCGngQsHfHJ7Gj6amQ) Content-type: text/x-vcard; charset=us-ascii; name=kmerriam.vcf Content-transfer-encoding: 7BIT Content-disposition: attachment; filename=kmerriam.vcf Content-description: Card for Kim Merriam 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 ------------------ - - --Boundary_(ID_cEnzMjvRYsdwRG03gELoNA) Content-type: text/plain; charset=US-ASCII Content-transfer-encoding: 7BIT <<<<<< See above "Message Body" >>>>>> - - --Boundary_(ID_cEnzMjvRYsdwRG03gELoNA) Content-type: text/html; charset=US-ASCII Content-transfer-encoding: 7BIT 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. ******************************************** - - ------------------ MIME Information follows ------------------ This is a multi-part message in MIME format. - - --Boundary_(ID_rAfeoYsSdfgjOYiaLx7+Eg) Content-type: text/plain; charset=us-ascii; x-mac-type=54455854; x-mac-creator=4D4F5353 Content-transfer-encoding: 7BIT <<<<<< See above "Message Body (Could be empty)" >>>>>> - - --Boundary_(ID_rAfeoYsSdfgjOYiaLx7+Eg) Content-type: message/rfc822 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 (ORCPT kmerriam@tktx.com); Tue, 24 Dec 2002 10:37:55 -0500 (EST) Received: (qmail 7929 invoked from network); Tue, 24 Dec 2002 15:39:14 +0000 Received: from unknown (HELO hobbes.ab.tac.net) (205.233.108.95) by bimini.th.net with SMTP; Tue, 24 Dec 2002 15:39:14 +0000 Received: (qmail 20408 invoked from network); Tue, 24 Dec 2002 15:39:13 +0000 Received: from unknown (HELO mail3.mail.cls.ab.ca) (208.38.55.67) by mail.ab.tac.net with SMTP; Tue, 24 Dec 2002 15:39:13 +0000 Received: by mail3.mail.cls.ab.ca with Internet Mail Service (5.5.2653.19) 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) Content-type: text/plain; charset=iso-8859-1 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 Content-disposition: attachment; filename=kmerriam.vcf Content-description: Card for Kim Merriam 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 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
**********************************************************************
          =^..^=
  - - --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 Content-transfer-encoding: 7BIT <<<<<< See above "Message Body" >>>>>> - --Boundary_(ID_HiNldVjQPh9t/PG78iTaug) Content-type: text/html; charset=US-ASCII Content-transfer-encoding: 7BIT 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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