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 [email protected] -----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of [email protected] Sent: Saturday, March 22, 2014 9:38 PM To: [email protected] 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? ----- Original Message ----- From: [email protected] To: [email protected] Subject: Histonet Digest, Vol 124, Issue 24 Send Histonet mailing list submissions to [email protected] To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to [email protected] You can reach the person managing the list at [email protected] When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. specimen marking ink (Davis, Cassie) 2. Leica Reichert Jung Cryocut 1800 (King, Laurie J) 3. RE: specimen marking ink ([email protected]) 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" <[email protected]> Subject: [Histonet] specimen marking ink To: "[email protected]" <[email protected]> Message-ID: <[email protected]> Content-Type: text/plain; charset="iso-8859-1" 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 [email protected] 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. ------------------------------ Message: 2 Date: Fri, 21 Mar 2014 17:45:48 +0000 From: "King, Laurie J" <[email protected]> Subject: [Histonet] Leica Reichert Jung Cryocut 1800 To: "'[email protected]' ([email protected])" <[email protected]> Message-ID: <[email protected]> Content-Type: text/plain; charset="us-ascii" Hello all, Looking for a manual for a Leica Reichert Jung Cryocut 1800. Laurie ______________________________________________________________________ The contents of this message may contain private, protected and/or privileged information. If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within. Please contact the sender and advise of the erroneous delivery by return e-mail or telephone. Thank you for your cooperation. ------------------------------ Message: 3 Date: Fri, 21 Mar 2014 14:37:59 -0500 From: <[email protected]> Subject: [Histonet] RE: specimen marking ink To: <[email protected]>, <[email protected]> Message-ID: <9e2d36ce2d7cba4a94d9b22e8328a3ba27efddf...@nadcwpmsgcms03.hca.corpad.net> Content-Type: text/plain; charset="iso-8859-1" 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: [email protected] [mailto:[email protected]] On Behalf Of Davis, Cassie Sent: Friday, March 21, 2014 1:04 PM To: [email protected] 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 [email protected] 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. _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 21 Mar 2014 13:06:46 -0700 (PDT) From: Jennifer Leigh <[email protected]> Subject: [Histonet] IHC on paraffin embedded skin tissue! To: Histonet Netserver <[email protected]> Message-ID: <[email protected]> Content-Type: text/plain; charset=iso-8859-1 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 <[email protected]> Subject: Re: [Histonet] RE: specimen marking ink To: "[email protected]" <[email protected]>, "[email protected]" <[email protected]>, "[email protected]" <[email protected]> Message-ID: <[email protected]> Content-Type: text/plain; charset=iso-8859-1 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, "[email protected]" <[email protected]> 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: [email protected] [mailto:[email protected]] On Behalf Of Davis, Cassie Sent: Friday, March 21, 2014 1:04 PM To: [email protected] 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 [email protected] 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. _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 21 Mar 2014 18:26:05 -0700 From: Bryan Llewellyn <[email protected]> Subject: Re: [Histonet] RE: specimen marking ink To: [email protected] Message-ID: <[email protected]> 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, "[email protected]" > <[email protected]> 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: [email protected] > [mailto:[email protected]] On Behalf Of Davis, Cassie > Sent: Friday, March 21, 2014 1:04 PM > To: [email protected] > 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 > [email protected] > 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. > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 7 Date: Fri, 21 Mar 2014 23:44:20 -0300 From: "C.D.G." <[email protected]> Subject: Re: [Histonet] IHC on paraffin embedded skin tissue! To: [email protected] Cc: [email protected] Message-ID: <[email protected]> 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 ------------------------------ _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 124, Issue 24 ***************************************** _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. 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