RE: [Histonet] Eosin Leaching

[Michael LaFriniere]

I have seen this happen with high levels of Isopropanol in alcohols after the 
eosins, I switch to regular reagent alcohols  and seemed to eliminate the 
bleeding of the eosin.

Michael
Michael R. LaFriniere, HT (ASCP) 
Executive Director
 

Capital Choice Pathology Laboratory
12041 Bournefield Way, Suite A * Silver Spring, MD 20904  
P: 240.471.3427 * F: 240.471.3401 * Cell 410-940-8844
michael.lafrini...@ccplab.com
 
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
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[Histonet] Eosin leaching

[Debra Siena]

Hi Histonet,


Eosin bleaching is usually due to the alcohols that are used after the eosin, 
if you are in a humid area, the alcohols can actually absorb water from the 
humidity in the air. This may mean a change of how often you rotate or change 
the alcohols, possibly more often than you did during the winter months when 
usually less humid.  Also if you use a graded alcohol after the eosin, maybe 
worth looking into going straight to 100%, I usually always recommend at least 
3 changes 100% alcohol for 1 minute and 3 changes of xylene for 1 minute.  If 
the last alcohol in your staining line is pink, it means that there is eosin 
and that means that there is water.  Eosin loves water and loves to come out of 
solution into the one containing the water.  I have even seen pink xylene in 
some staining lines due to water in xylene, however, not enough that it has 
turned cloudy yet but still have a bit of water.  Xylene will tolerate about 
.5% water before becoming too milky or cloudy to use.  Also when you rotate 
your solutions make sure that you dry the containers well before  refilling and 
by graduating the levels of the alcohols and xylenes to make sure that your 
last container completely covers the slides. Lastly, adding a 95% alcohol 
before your eosin, if you don't have one may help to keep excess water out of 
your eosin which will cause the eosin to break down faster, again eosin loves 
water.  Just spouting off the top of my head but hope something helps out.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

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RE: [Histonet] Eosin Leaching

[Tony Reilly]

Hi Adrienne

Also make sure the wash after your bluing agent is higher than the blueing 
solution or alkalinity of the solution will leach out your eosin.

Regards
Tony


Tony Reilly B.App.Sc,  M.Sc
Chief Scientist
Anatomical Pathology
Pathology Queensland PAH
_
Health Services Support Agency| Department of Health

Building 15, Level 1, 
199 Ipswich Road 
WOOLLOONGABBA  Queensland 4102
Ph: 07 3176 2412
Mob: 0402139411
Fax: 07 3176 2930
Email: tony.reil...@health.qld.gov.au | www.health.qld.gov.au 
     





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Friday, 20 June 2014 10:11 PM
To: Sanders, Jeanine (CDC/OID/NCEZID); 'Adrienne Anderson'; 
'histo...@listsutsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Is your alcohol level as high as or higher than the level of eosin in the 
containers? If there is residual eosin left at the upper portion of the slide, 
that could be causing the problem.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
Jeanine (CDC/OID/NCEZID)
Sent: Thursday, June 19, 2014 5:26 PM
To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
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RE: [Histonet] Eosin Leaching

[Vanessa Avalos]

We had the same problem. Increase your Alcohol/Reagents (following Eosin) times 
as well as using fresh ones. I use 80% for about 20 sec and (2)100% for a 
minute each and (3) Xylene Subs for 2 min each. I also use a fresh 80% and 
rotate the 100% each day. If using a Xylene Sub it could also be a possibility 
that your mounting medium is not compatible. 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 2:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
or may contain health information protected by state and federal law.
Information and opinions expressed in this message and/or attachments are those 
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RE: [Histonet] Eosin Leaching

[Anita Buchiane]

I'd advise checking your dehydration alcohols with a hydrometer.  This 
"bleeding" happened to me once and it turned out the alcohols after the Eosin 
were not graded.  My lab aide accidently used 100% only instead of 70%, 95% and 
then 100%.  
Anita

Anita

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Friday, June 20, 2014 8:15 AM
To: Rathborne, Toni
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Eosin Leaching

We haven't changed our mounting medium, but I'll have to check on the alcohol 
levels. When I stain (we hand stain only - very small lab), I always make sure 
the alcohol is higher than the eosin, but I have a lab assistant now, and it's 
very possible I didn't make that a point. I'll try that.

Any more tips are welcome still - thank you all!

Adrienne


On Jun 20, 2014, at 8:11 AM, Rathborne, Toni  wrote:

> Is your alcohol level as high as or higher than the level of eosin in the 
> containers? If there is residual eosin left at the upper portion of the 
> slide, that could be causing the problem.
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
> Jeanine (CDC/OID/NCEZID)
> Sent: Thursday, June 19, 2014 5:26 PM
> To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
> Subject: RE: [Histonet] Eosin Leaching
> 
> Have you changed your coverslipping mountant?
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
> Anderson
> Sent: Thursday, June 19, 2014 5:24 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Eosin Leaching
> 
> Hello all,
> 
> We're having a problem with our eosin bleeding out of the sections. I've read 
> that this can be caused by not dehydrating adequately after eosin, but we're 
> still having an issue when using fresh alcohols. Does anyone have any other 
> ideas as to the cause of this problem?
> 
> Thanks in advance for your help!
> 
> Adrienne
> 
> 
> 
> Adrienne Anderson, BS, HTL(ASCP)
> Histotechnologist
> Phylogeny, Inc. 
> 1476 Manning Pkwy, Powell, Ohio 43605
> Phone: (614) 846-6161 
> Fax:  (877) 591-1815
> 
> This message, including any attachments, is confidential and may be 
> privileged or may contain health information protected by state and federal 
> law.
> Information and opinions expressed in this message and/or attachments are 
> those of the author and are not necessarily those of the company. 
> If you are not the intended recipient, please notify the sender and delete 
> this message from your system. 
> Any use of this information by individuals other than the intended recipient 
> is strictly prohibited. ___
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Re: [Histonet] Eosin Leaching

[Adrienne Anderson]

We haven’t changed our mounting medium, but I’ll have to check on the alcohol 
levels. When I stain (we hand stain only - very small lab), I always make sure 
the alcohol is higher than the eosin, but I have a lab assistant now, and it’s 
very possible I didn’t make that a point. I’ll try that.

Any more tips are welcome still - thank you all!

Adrienne


On Jun 20, 2014, at 8:11 AM, Rathborne, Toni  wrote:

> Is your alcohol level as high as or higher than the level of eosin in the 
> containers? If there is residual eosin left at the upper portion of the 
> slide, that could be causing the problem.
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
> Jeanine (CDC/OID/NCEZID)
> Sent: Thursday, June 19, 2014 5:26 PM
> To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
> Subject: RE: [Histonet] Eosin Leaching
> 
> Have you changed your coverslipping mountant?
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
> Anderson
> Sent: Thursday, June 19, 2014 5:24 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Eosin Leaching
> 
> Hello all,
> 
> We're having a problem with our eosin bleeding out of the sections. I've read 
> that this can be caused by not dehydrating adequately after eosin, but we're 
> still having an issue when using fresh alcohols. Does anyone have any other 
> ideas as to the cause of this problem?
> 
> Thanks in advance for your help!
> 
> Adrienne
> 
> 
> 
> Adrienne Anderson, BS, HTL(ASCP)
> Histotechnologist
> Phylogeny, Inc. 
> 1476 Manning Pkwy, Powell, Ohio 43605
> Phone: (614) 846-6161 
> Fax:  (877) 591-1815
> 
> This message, including any attachments, is confidential and may be 
> privileged or may contain health information protected by state and federal 
> law.
> Information and opinions expressed in this message and/or attachments are 
> those of the author and are not necessarily those of the company. 
> If you are not the intended recipient, please notify the sender and delete 
> this message from your system. 
> Any use of this information by individuals other than the intended recipient 
> is strictly prohibited. ___
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> Histonet@lists.utsouthwestern.edu
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> 
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RE: [Histonet] Eosin Leaching

[Rathborne, Toni]

Is your alcohol level as high as or higher than the level of eosin in the 
containers? If there is residual eosin left at the upper portion of the slide, 
that could be causing the problem.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
Jeanine (CDC/OID/NCEZID)
Sent: Thursday, June 19, 2014 5:26 PM
To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
or may contain health information protected by state and federal law.
Information and opinions expressed in this message and/or attachments are those 
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[Histonet] Eosin Leaching

[Adrienne Anderson]

Hello all,

We’re having a problem with our eosin bleeding out of the sections. I’ve read 
that this can be caused by not dehydrating adequately after eosin, but we’re 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
or may contain health information protected by state and federal law.
Information and opinions expressed in this message and/or attachments are those 
of the author and are not necessarily those of the company. 
If you are not the intended recipient, please notify the sender and delete this 
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RE: [Histonet] Eosin Leaching

[Sanders, Jeanine (CDC/OID/NCEZID)]

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
or may contain health information protected by state and federal law.
Information and opinions expressed in this message and/or attachments are those 
of the author and are not necessarily those of the company. 
If you are not the intended recipient, please notify the sender and delete this 
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RE: [Histonet] Eosin leaching out of sections

[Rathborne, Toni]

Where do you get the beads from?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Joyce
Cline
Sent: Wednesday, February 03, 2010 2:02 PM
To: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Eosin leaching out of sections 


Humidity has played a large part in our slides leaching Eosin. We have a 
dehumidifier in our staining room and we also use H2Blue Beads in our Xylene 
substitute. The beads absorb water from the substitute and help prevent 
leaching of the stain.

The beads are ordered from Amercian Mastertech.

Joyce Cline, Technical Specialist
Hagerstown Medical Laboratory
301-665-4980
fax 301-665-4941

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections

Hi again,

Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide?

Thanks,

Scott Hendricksen HT(ASCP)
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RE: [Histonet] Eosin leaching out of sections

[Joyce Cline]

Humidity has played a large part in our slides leaching Eosin. We have a 
dehumidifier in our staining room and we also use H2Blue Beads in our Xylene 
substitute. The beads absorb water from the substitute and help prevent 
leaching of the stain.

The beads are ordered from Amercian Mastertech.

Joyce Cline, Technical Specialist
Hagerstown Medical Laboratory
301-665-4980
fax 301-665-4941

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections

Hi again,

Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide?

Thanks,

Scott Hendricksen HT(ASCP)
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FW: [Histonet] Eosin leaching out of sections

[Ian Montgomery]

Scott,
Did you mount from water or a clearing agent?

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections 

Hi again,

Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide? 

Thanks,

Scott Hendricksen HT(ASCP)
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RE: [Histonet] Eosin leaching out of sections

[Bonner, Janet]

Your xylene and/or alcohols may need changing.
 
Janet 



From: histonet-boun...@lists.utsouthwestern.edu on behalf of Scott Hendricksen
Sent: Wed 2/3/2010 9:02 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections 



> Hi again,
>
> Does anyone have an issue with eosin leaching out of sections a few days 
> after they have been coverslipped on an H&E stained slide?
>
> Thanks,
>
> Scott Hendricksen HT(ASCP)


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[Histonet] Eosin leaching out of sections

[Scott Hendricksen]

> Hi again,
> 
> Does anyone have an issue with eosin leaching out of sections a few days 
> after they have been coverslipped on an H&E stained slide? 
> 
> Thanks,
> 
> Scott Hendricksen HT(ASCP)


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[Histonet] Eosin leaching out of sections

[Scott Hendricksen]

Hi again,

Does anyone have an issue with eosin leaching out of sections a few days after 
they have been coverslipped on an H&E stained slide? 

Thanks,

Scott Hendricksen HT(ASCP)
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[Histonet] Eosin leaching out of sections

[Scott Hendricksen]

Hi again,

Does anyone have an issue with eosin leaching out of sections a few days after 
they have been coverslipped on an H&E stained slide? 

Thanks,

Scott Hendricksen HT(ASCP)
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