[Histonet] RE: Help kappa and lambda IHC on bone marrow bxs
Cartun, Richard
Richard.Cartun <@t> hhchealth.org
Mon Mar 24 14:50:43 CDT 2014
I would recommend using higher dilutions of your primary antibodies (try 1:20,000). As you might expect, it's not unusual to see a lot of background immunoglobulin immunoreactivity in bone marrow specimens. We actually have 2 antibody dilutions for both Kappa and Lambda that we run on the Leica Bond Max; a routine dilution and then a higher one for these types of situations.
Richard
Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 972-1596 Office
(860) 545-2204 Fax
richard.cartun <@t> hhchealth.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of drmoses111 <@t> comcast.net
Sent: Saturday, March 22, 2014 9:38 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Help kappa and lambda IHC on bone marrow bxs
Our lab has started using Immunocal decal solution on our bone marrows. Most of our antibodies have improved except kappa and lambda. We do not do ISH. Kappa and lambda staining in the tonsil controls is good, The bone marrows are now very overstrained. We use DAKO polyclonals at 1:10,000 with protease1 on the Ventana Ultra. Does anyone have a procedure?
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Subject: Histonet Digest, Vol 124, Issue 24
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Today's Topics:
1. specimen marking ink (Davis, Cassie)
2. Leica Reichert Jung Cryocut 1800 (King, Laurie J)
3. RE: specimen marking ink (Wanda.Smith <@t> HCAhealthcare.com)
4. IHC on paraffin embedded skin tissue! (Jennifer Leigh)
5. Re: RE: specimen marking ink (David Kemler)
6. Re: RE: specimen marking ink (Bryan Llewellyn)
7. Re: IHC on paraffin embedded skin tissue! (C.D.G.)
----------------------------------------------------------------------
Message: 1
Date: Fri, 21 Mar 2014 13:03:35 -0400
From: "Davis, Cassie" <CDavis <@t> che-east.org>
Subject: [Histonet] specimen marking ink
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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<BE7AF02009C3A74F9B8344E70F3655CBFB9B2B <@t> CHEXCMS01.one.ads.che.org>
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Hi Histo World, as I was cutting to day I was thinking why don't we see if we could get specimen marking ink directly from a tattoo vendor? When I first started in histo I was told the ink we use was actually tattoo ink. As we know as soon as somebody labels something as a "medical supply" the price is increased. Just a cost saving thought, I mentioned it to my immediate supervisor but she think it would be a liability issue. I thought we could "test/validate it on skin tissue left over from a mastectomy or extremity. Any thoughts?
Cassandra Davis
CDavis <@t> che-east.org
302-575-8095
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Message: 2
Date: Fri, 21 Mar 2014 17:45:48 +0000
From: "King, Laurie J" <king.laurie <@t> marshfieldclinic.org>
Subject: [Histonet] Leica Reichert Jung Cryocut 1800
To: "'histonet <@t> lists.utsouthwestern.edu'
(histonet <@t> lists.utsouthwestern.edu)"
<histonet <@t> lists.utsouthwestern.edu>
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Hello all,
Looking for a manual for a Leica Reichert Jung Cryocut 1800.
Laurie
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Message: 3
Date: Fri, 21 Mar 2014 14:37:59 -0500
From: <Wanda.Smith <@t> HCAhealthcare.com>
Subject: [Histonet] RE: specimen marking ink
To: <CDavis <@t> che-east.org>, <histonet <@t> lists.utsouthwestern.edu>
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Back in the 70's (when I was six, but still a histotech!!!) my Pathologist had gotten tattoo pigment powder in two colors and that's what we used to mark tissue. Don't know where it came from and don't know where it went!!!
Happy Friday and Happy Weekend to everyone!!!
Wanda
WANDA G. SMITH, HTL(ASCP)HT
Pathology Supervisor
TRIDENT MEDICAL CENTER
9330 Medical Plaza Drive
Charleston, SC 29406
843-847-4586
843-847-4296 fax
This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Davis, Cassie
Sent: Friday, March 21, 2014 1:04 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] specimen marking ink
Hi Histo World, as I was cutting to day I was thinking why don't we see if we could get specimen marking ink directly from a tattoo vendor? When I first started in histo I was told the ink we use was actually tattoo ink. As we know as soon as somebody labels something as a "medical supply" the price is increased. Just a cost saving thought, I mentioned it to my immediate supervisor but she think it would be a liability issue. I thought we could "test/validate it on skin tissue left over from a mastectomy or extremity. Any thoughts?
Cassandra Davis
CDavis <@t> che-east.org
302-575-8095
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Message: 4
Date: Fri, 21 Mar 2014 13:06:46 -0700 (PDT)
From: Jennifer Leigh <immunoqueen <@t> yahoo.com>
Subject: [Histonet] IHC on paraffin embedded skin tissue!
To: Histonet Netserver <histonet <@t> lists.utsouthwestern.edu>
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Histonetters-
I have a question about mouse skin samples----an IHC experiment has gone wrong and I am not sure why. I have a set of paraffin embedded mouse skin samples from an inflammatory model that were stained for F4/80. The end result is no color development after the addition of DAB. I use the DAB tablets from Sigma using the TBS buffer to dissolve the tablet. There is no color development, not even non-specific staining. Any ideas what the problem might be????? I also performed a Proteinase K antigen retrevial step (using the ready made solution from Dako) for 15 minutest at 37 C as I was instructed to do. Thank you in advance for any insight!
Jennifer Oskins
Jennifer L. Oskins
"Until one has loved an animal, part of their soul remains unawakened....."
------------------------------
Message: 5
Date: Fri, 21 Mar 2014 16:02:29 -0700 (PDT)
From: David Kemler <histotalk <@t> yahoo.com>
Subject: Re: [Histonet] RE: specimen marking ink
To: "Wanda.Smith <@t> HCAhealthcare.com" <Wanda.Smith <@t> HCAhealthcare.com>,
"CDavis <@t> che-east.org" <CDavis <@t> che-east.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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Actually, I have a tattoo supplier four miles away from me. Several years ago, I went to their warehouse to take a look an possibly make a purchase. As it turned out, I had to buy a gallon of each color and it worked out to be more than the standard tissue inks sold by the histology vendors.
Next stop - tattoo shop. Same story. The artist would sell a small 8 oz. bottle at cost, but it was still just a bit more than our vendors.
You can use Pelican drawing ink. It comes in several colors. It's around five something for an ounce+, much more than Davidson or one of the others.
Maybe someone will also chime in.
Dave
On Friday, March 21, 2014 3:39 PM, "Wanda.Smith <@t> HCAhealthcare.com" <Wanda.Smith <@t> HCAhealthcare.com> wrote:
Back in the 70's (when I was six, but still a histotech!!!) my Pathologist had gotten tattoo pigment powder in two colors and that's what we used to mark tissue. Don't know where it came from and don't know where it went!!!
Happy Friday and Happy Weekend to everyone!!!
Wanda
WANDA G. SMITH, HTL(ASCP)HT
Pathology Supervisor
TRIDENT MEDICAL CENTER
9330 Medical Plaza Drive
Charleston, SC 29406
843-847-4586
843-847-4296 fax
This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Davis, Cassie
Sent: Friday, March 21, 2014 1:04 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] specimen marking ink
Hi Histo World, as I was cutting to day I was thinking why don't we see if we could get specimen marking ink directly from a tattoo vendor? When I first started in histo I was told the ink we use was actually tattoo ink. As we know as soon as somebody labels something as a "medical supply" the price is increased. Just a cost saving thought, I mentioned it to my immediate supervisor but she think it would be a liability issue. I thought we could "test/validate it on skin tissue left over from a mastectomy or extremity. Any thoughts?
Cassandra Davis
CDavis <@t> che-east.org
302-575-8095
Confidentiality Notice:
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------------------------------
Message: 6
Date: Fri, 21 Mar 2014 18:26:05 -0700
From: Bryan Llewellyn <llewllew <@t> shaw.ca>
Subject: Re: [Histonet] RE: specimen marking ink
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <532CE6AD.8050806 <@t> shaw.ca>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
You can buy tattoo ink in a bewildering array of colours from ebay quite
cheaply.
Many years ago I tried to buy tattoo ink from a tattoo parlour, but they
refused to sell it to me as they were concerned that it would be used
for rubbing into scratches and cuts for home made tattoos. I was in my
mid-50's at the time. Perhaps I looked strange with my beard and long
hair! I decided to buy the commercial tattoo ink kits from lab suppliers
for mixed colours, but used laundry blueing for most margins and kept
the tattoo inks for those occasions when it was important to know which
margin was which.
Bryan Llewellyn
David Kemler wrote:
> Actually, I have a tattoo supplier four miles away from me. Several years ago, I went to their warehouse to take a look an possibly make a purchase. As it turned out, I had to buy a gallon of each color and it worked out to be more than the standard tissue inks sold by the histology vendors.
>
> Next stop - tattoo shop. Same story. The artist would sell a small 8 oz. bottle at cost, but it was still just a bit more than our vendors.
>
> You can use Pelican drawing ink. It comes in several colors. It's around five something for an ounce+, much more than Davidson or one of the others.
>
> Maybe someone will also chime in.
>
> Dave
>
>
>
> On Friday, March 21, 2014 3:39 PM, "Wanda.Smith <@t> HCAhealthcare.com" <Wanda.Smith <@t> HCAhealthcare.com> wrote:
>
> Back in the 70's (when I was six, but still a histotech!!!) my Pathologist had gotten tattoo pigment powder in two colors and that's what we used to mark tissue. Don't know where it came from and don't know where it went!!!
> Happy Friday and Happy Weekend to everyone!!!
> Wanda
>
> WANDA G. SMITH, HTL(ASCP)HT
> Pathology Supervisor
> TRIDENT MEDICAL CENTER
> 9330 Medical Plaza Drive
> Charleston, SC 29406
> 843-847-4586
> 843-847-4296 fax
>
> This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed.
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Davis, Cassie
> Sent: Friday, March 21, 2014 1:04 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] specimen marking ink
>
> Hi Histo World, as I was cutting to day I was thinking why don't we see if we could get specimen marking ink directly from a tattoo vendor? When I first started in histo I was told the ink we use was actually tattoo ink. As we know as soon as somebody labels something as a "medical supply" the price is increased. Just a cost saving thought, I mentioned it to my immediate supervisor but she think it would be a liability issue. I thought we could "test/validate it on skin tissue left over from a mastectomy or extremity. Any thoughts?
>
> Cassandra Davis
> CDavis <@t> che-east.org
> 302-575-8095
>
>
>
> Confidentiality Notice:
> This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s).
> It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email.
>
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------------------------------
Message: 7
Date: Fri, 21 Mar 2014 23:44:20 -0300
From: "C.D.G." <latecor <@t> montevideo.com.uy>
Subject: Re: [Histonet] IHC on paraffin embedded skin tissue!
To: Histonet <@t> lists.utsouthwestern.edu
Cc: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <201403212344200234.000D0792 <@t> smtp.montevideo.com.uy>
Content-Type: text/plain; charset="ISO-8859-1"
Jennifer be sure to add three or four drops of H2O2 to the solution. Maybe the lack
of oxidation of the chromogen is the cause of no color development.
Best luck,
Carlos.-
> Histonetters-
I have a question about mouse skin
>samples----an IHC experiment has gone wrong and I am not sure why. I have
>a set of paraffin embedded mouse skin samples from an inflammatory model
>that were stained for F4/80. The end result is no color development after
>the addition of DAB. I use the DAB tablets from Sigma using the TBS buffer
>to dissolve the tablet. There is no color development, not even
>non-specific staining. Any ideas what the problem might be????? I also
>performed a Proteinase K antigen retrevial step (using the ready made
>solution from Dako) for 15 minutest at 37 C as I was instructed to do.
>Thank you in advance for any insight!
Jennifer Oskins
Jennifer L.
>Oskins
"Until one has loved an animal, part of their soul remains
>unawakened....."
>_______________________________________________
>Histonet mailing list
>
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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