From adesupo2002 at hotmail.com Tue Sep 23 12:39:52 2025 From: adesupo2002 at hotmail.com (ADESUPO ADESUYI) Date: Tue, 23 Sep 2025 17:39:52 +0000 Subject: [Histonet] Performing IHC stain on an H&E stained slides Message-ID: Hi, Does anyone has a protocol for performing IHC stain on an H&E stained slides to share? Sincerely, Ade From adesupo2002 at hotmail.com Tue Sep 23 12:48:52 2025 From: adesupo2002 at hotmail.com (ADESUPO ADESUYI) Date: Tue, 23 Sep 2025 17:48:52 +0000 Subject: [Histonet] Performing IHC stain on an H&E stained slides In-Reply-To: References: Message-ID: Hi, Does anyone have a protocol for performing IHC stain on an H&E stained slides to share? Sincerely, Ade From Jessica.Piche at wtbyhosp.org Wed Sep 24 08:29:03 2025 From: Jessica.Piche at wtbyhosp.org (Piche, Jessica) Date: Wed, 24 Sep 2025 13:29:03 +0000 Subject: [Histonet] Renal biopsy charges Message-ID: Good Morning, I have a question regarding billing for a renal biopsy. When we receive a renal biopsy we process, embedd, and cut the block. We make 16 slides. Two for H&E, one of which is sent with the 14 unstained slides to another facility with the EM and IF portion of the renal biopsy. Our pathologist gets one H&E. Our pathologist does not make the diagnosis. How should this be billed? We are currently billing 88305 for the technical component. I cannot find a direct answer on how this should be billed. Does anyone know? Thank you and have a great day. Jessica Jessica Pich?, HT(ASCP) Histology Team Leader, Laboratory Waterbury Health 64 Robbins Street Waterbury, Connecticut, 06708 Phone: 203-573-7167 FAX: 203-573-7242 Email:jessica.piche at wtbyhosp.org Waterbury HEALTH From histo at pathlab.us Wed Sep 24 09:45:05 2025 From: histo at pathlab.us (Histology) Date: Wed, 24 Sep 2025 14:45:05 +0000 Subject: [Histonet] Performing IHC stain on an H&E stained slides In-Reply-To: References: Message-ID: <2e3eff1307b94c6ba035be2d1967aea4@pathlab.us> We do this often. We take the coverslip off and put the slide in xylene for at least 5 minutes. Then, we run it down to water. From the water, we load it on our Ventana ultra with the same protocol as normal but without the deparaffination step. They have an option for "wet-load". I'm sure this can be done on other platforms and manually as well. Mehndi Helgren Lab Manager 757-664-7901 Dominion Pathology Labs. 733 Boush St. Suite 200 Norfolk, VA ?23510 -----Original Message----- From: ADESUPO ADESUYI via Histonet Sent: Tuesday, September 23, 2025 1:40 PM To: histonet at lists.utsouthwestern.edu Subject: [Histonet] Performing IHC stain on an H&E stained slides Hi, Does anyone has a protocol for performing IHC stain on an H&E stained slides to share? Sincerely, Ade _______________________________________________ Histonet mailing list Histonet at lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From suefaso at gmail.com Wed Sep 24 14:00:20 2025 From: suefaso at gmail.com (Sue Faso) Date: Wed, 24 Sep 2025 15:00:20 -0400 Subject: [Histonet] Clinical trials support Message-ID: Hello Histonetters, How are your organizations handling blocks/slides requested for clinical trials? Do you send blocks (usually preferred by sponsors), or sections? Can anyone share how much they charge for sectioning slides for clinical trials? Thanks, Sue Susan Faso Laboratory Coordinator Pathology Core Research Lab SUNY Upstate Syracuse, NY Fasos <@t> upstate.edu Office/Lab 315-464-4812 From amosbrooks at gmail.com Wed Sep 24 16:17:11 2025 From: amosbrooks at gmail.com (Amos Brooks) Date: Wed, 24 Sep 2025 17:17:11 -0400 Subject: [Histonet] Performing IHC stain on an H&E stained slides In-Reply-To: References: Message-ID: Hi, Soak the coverslip off and let it sit in xylene long enough to make absolutely sure all the mounting media is dissolved. Then starting from 100% ETOH rehydrate to water then buffer. And start the IHC as usual. There is no need to destain. The ethanol will remove the eosin and you are likely to counterstain with hematoxylin anyway. Make sure you do repeat the counterstain as well since the hematoxylin will probably bleach out if not entirely, certainly some. Amos > Message: 1 > Date: Tue, 23 Sep 2025 17:39:52 +0000 > From: ADESUPO ADESUYI > To: "histonet at lists.utsouthwestern.edu" > > Subject: [Histonet] Performing IHC stain on an H&E stained slides > Message-ID: > < > BY5PR17MB3923E870CDD791A61CF82A99B61DA at BY5PR17MB3923.namprd17.prod.outlook.com > > > > Content-Type: text/plain; charset="iso-8859-1" > > Hi, > > Does anyone has a protocol for performing IHC stain on an H&E stained > slides to share? > > Sincerely, > Ade From jdhannasch at gmail.com Wed Sep 24 17:31:11 2025 From: jdhannasch at gmail.com (jdhannasch at gmail.com) Date: Wed, 24 Sep 2025 15:31:11 -0700 Subject: [Histonet] Performing IHC stain on an H&E stained slides In-Reply-To: <2e3eff1307b94c6ba035be2d1967aea4@pathlab.us> References: <2e3eff1307b94c6ba035be2d1967aea4@pathlab.us> Message-ID: I believe the ventana stainer does the antigen retrieval step as part of their protocol. If someone is staining by hand or has a stainer that does not do the antigen retrieval step, they will need to make sure to do it before staining. > On Sep 24, 2025, at 7:45?AM, Histology wrote: > > ?We do this often. We take the coverslip off and put the slide in xylene for at least 5 minutes. Then, we run it down to water. From the water, we load it on our Ventana ultra with the same protocol as normal but without the deparaffination step. They have an option for "wet-load". I'm sure this can be done on other platforms and manually as well. > > > Mehndi Helgren > Lab Manager > 757-664-7901 > Dominion Pathology Labs. > 733 Boush St. Suite 200 > Norfolk, VA 23510 > > > > -----Original Message----- > From: ADESUPO ADESUYI via Histonet > Sent: Tuesday, September 23, 2025 1:40 PM > To: histonet at lists.utsouthwestern.edu > Subject: [Histonet] Performing IHC stain on an H&E stained slides > > Hi, > > Does anyone has a protocol for performing IHC stain on an H&E stained slides to share? > > Sincerely, > Ade > _______________________________________________ > Histonet mailing list > Histonet at lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From edmartin26 at gmail.com Thu Sep 25 18:12:39 2025 From: edmartin26 at gmail.com (Eddie Martin) Date: Thu, 25 Sep 2025 19:12:39 -0400 Subject: [Histonet] Help To: Handling Blocks/slides requested for Clinical Trials Histonet Digest, Vol 262, Issue 2 In-Reply-To: References: Message-ID: Hello Sue, I work in the Bone Marrow Service Laboratory at the NIH. Each facility should review its own laboratory policies to determine what materials may or may not be sent out for clinical trials. At our facility, all patients are either being evaluated for, or are already enrolled in, a clinical trial. Following their care at the NIH, patients may require tissue slides generated here to be forwarded to the facility responsible for their continued care. Regarding charges for preparing slides: I am not aware of any billing to patients or to the clinical research study group overseeing their trial. Below is a summary of the materials we typically request clinical coordinators to communicate to sending institutions. The final decision on what is shipped rests with the sending institution: - The original tissue block - For Bone Marrow Core & Aspirate Clot: Ten charged slides, each with one unstained section of FFPE tissue. - For bone marrow pathology: any remaining unstained aspirate, touch imprint, and peripheral blood smear referenced in the diagnostic report. - Our services works only with bone marrow tissue. However, in my past work experiences, other anatomic tissues also benefit in niche circumstances if a few unstained slides shipped have 2-3 sequential sections to rule out questionable morphology. *Notes:* 1. Depending on the institution receiving the materials, tissue blocks or stained slides may be retained for a defined period after a case has been electronically signed out. In such cases, particularly if the patient's materials are needed sooner for their continuation of care, timely communication with the clinical trial coordinator is needed to expedite the shipping back of original materials received. 2. Clinical trial testing requirements for materials needed will vary depending on the specific protocol the patient is being admitted to. 3. In rare situations, additional material may be requested, where either the slides shipped aren't charged, or the brand of charged slides isn't compatible with the IHC instrumentation where the clinical study is being performed. I am happy to respond to further questions if needed. My contact information is included in my email signature. Best regards, Eddie Martin Technical Pathology Specialist The National Institutes of Health 10 Center Drive Building 10, RM 2C301 Bethesda, MD 20816 (301) 594-2054 eddie.martin at nih.gov > ---------- Forwarded message ---------- > From: Sue Faso > To: histonet at lists.utsouthwestern.edu > Cc: > Bcc: > Date: Wed, 24 Sep 2025 15:00:20 -0400 > Subject: [Histonet] Clinical trials support > Hello Histonetters, > > How are your organizations handling blocks/slides requested for clinical > trials? > Do you send blocks (usually preferred by sponsors), or sections? > > Can anyone share how much they charge for sectioning slides for clinical > trials? > > Thanks, > Sue > > Susan Faso > Laboratory Coordinator > Pathology Core Research Lab > SUNY Upstate > Syracuse, NY > Fasos <@t> upstate.edu > Office/Lab 315-464-4812 > > > > > From edmartin26 at gmail.com Thu Sep 25 20:19:49 2025 From: edmartin26 at gmail.com (Eddie Martin) Date: Thu, 25 Sep 2025 21:19:49 -0400 Subject: [Histonet] Help to: Nuclear Fast Red Counterstain (Eddie Martin) Histonet Digest, Vol 261, Issue 5 In-Reply-To: References: Message-ID: > > Hello Mehndi, > > Our lab hasn?t experienced this in some time?generally its resolved with > adding extra online washes with surfactant post NFR counterstain. There are > easily 3-4 practices done in a high volume histolab that may easily > exacerbate the crystals you?re mentioning. Common practices such as: > > ? letting slides dry on the special stain instrumentation > > ? dehydrating too quickly after offline DI water rinses > > ? Oven drying above 45C > > ? The histolab?s humidity is greater than 60% > > Assuming the NFR commercially available reagent is properly made at an > appropriate concentration of NFR, and your laboratory?s humidity level is > between 40% - 60%, and the slides counterstained with NFR weren?t left to > dry on the special stain instrument or when performed manually, then the > following can be done to remove residual NFR crystals from embedding onto > the slide: > > Note: If your Special Stain instrumentation has a slide drying process at > the end of the run, disable it and perform the following offline steps > while the slides are still wet after the rinsing steps on the Special Stain > protocol: > > 1. Tween 20 rinse (0.05?1.0%): 1 min with gentle agitation. This will > remove loose precipitate left after NFR is rinsed off. > 2. Acetic dip (optional yet highly suggested) : A few dips in 1% > glacial acetic acid. An acetic acid dip protonates residual dye. Any > residual crystals, if present on the wet slide have another opportunity to > be removed by charging the precipitated crystals to enable them to be > rinsed off the slides. Rinses in DI water is necessary to stop the acidity > effect. > > ? If skipping acetic dip: go straight to DI water rinses after > Tween 20 rinse. > > 3. Graded alcohols ? Quick passes only: > > ? 70%: 15?30 sec > > ? 80%: 15 sec > > ? 95%: 15?30 sec > > ? 100%: 15?30 sec (?2) > > ? Caution: Excess time in alcohol bleaches NFR. > > ? Caution: Skipping graded alcohols to get to 100% alcohol sooner > also causes precipitate to form. > > > 4. Xylene: Extend time in the second xylene if the first xylene is not > changed frequently. (this avoids accidental bleaching in first xylene with > excess residual 100% alcohol). > 5. Oven drying: if choosing to oven dry instead of dehydrating through > graded alcohols; Slide drying with warm temps between 40 ?C & 60 ?C. > Slide oven temps >60 ?C may also cause uneven drying and crystal > precipitation. > 6. Other factors previously mentioned above: Over-concentrated NFR > either commercially available or when making a working solution, high > humidity >65%, and shortcuts taken in graded alcohols or elevating oven > temps above 40 ?C each individually contribute to crystals embedding onto > the slide. > > > I hope this helps! > > Eddie Martin > Technical Pathology Specialist > The National Institutes of Health > 10 Center Drive > Building 10, RM 2C301 > Bethesda, MD 20816 > (301) 594-2054 > eddie.martin at nih.gov > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > Today's Topics: > > 1. Nuclear fast red counterstain (Histology) > 2. Re: Nuclear fast red counterstain (John Kiernan) > > > > ---------- Forwarded message ---------- > From: Histology > To: Histonet > Cc: > Bcc: > Date: Tue, 19 Aug 2025 17:31:00 +0000 > Subject: [Histonet] Nuclear fast red counterstain > Hi all, > > Has anyone experienced commercially pre-made Nuclear Fast Red having > reddish crystals in it? We use this as a counterstain for Prussian Iron > and Melanin stains and we are seeing little crystals on the slide. We have > tried filtering the solution but we are still seeing this problem. Any > help would be much appreciated. > > Thanks as always Histonet!! > > Mehndi Helgren > Lab Manager > 757-664-7901 > Dominion Pathology Labs. > 733 Boush St. Suite 200 > Norfolk, VA 23510 > > > From edmartin26 at gmail.com Thu Sep 25 21:32:54 2025 From: edmartin26 at gmail.com (Eddie Martin) Date: Thu, 25 Sep 2025 22:32:54 -0400 Subject: [Histonet] Help to: Changing Alcohol Vendors. Histonet Digest, Vol 261, Issue 3 In-Reply-To: References: Message-ID: > > Good evening, > > > > At the end of the day, your SOP should spell out what to do in this > situation. Let that document, signed off by your technical manager or > director, guide the call. > > Here?s my take (strictly my opinion, not your lab?s): > > If you?re using a pure solvent like ethanol or isopropanol, or Xylene. > the CAS number doesn?t change by vendor. > > 1. If the reagent is part of the stain?s analytical method: (H&E > stain), then a small comparison study is usually all that?s needed since > the reagents being switched are exactly the same in their properties & > concentrations. The only change is the distributor/vendor the reagent is > being acquired from. > > 2. If the reagent is *Not* part of the stain?s analytical method: > If the alcohol and xylene aren?t method steps for performing the test > (such as being the dehydration and clearing steps only) then I wouldn?t > consider that revalidation-worthy. > > > > I hope this helps! > > > > Best regards, > Eddie Martin > Technical Pathology Specialist > The National Institutes of Health > 10 Center Drive > Building 10, RM 2C301 > Bethesda, MD 20816 > (301) 594-2054 > eddie.martin at nih.gov > > On Wed, Jul 23, 2025, 11:44?AM Pairan, Kelly via Histonet < > histonet at lists.utsouthwestern.edu> wrote: > > > Good Morning, > > Our hospital is looking at switching our 100% alcohol and 95% alcohol to > > another vendor. I have received mixed answers on whether or not we are > > going to need to a validation for this reagent swap when it comes to > > processing, staining and IHC. What is the practice at your institution? > > > > Thanks, > > Kelly > > > > Kelly Pairan, HT(ASCP)CM, HQIPCM > > Technical Scientist Anatomic Pathology and Cytology > > OhioHealth Laboratory Systems > > Suite 210 North Medical Building > > 3535 Olentangy River Rd > > Columbus, OH 43214 > > > > Email: kelly.pairan at ohiohealth.com > > Work: (614) 566-3575 > > Cell: (614) 312-0104 > > > > _______________________________________________ > > Histonet mailing list > > Histonet at lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet at lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet