Re: [Histonet] processing artifact - delayed start
I am wondering if you are getting air bubbles/pockets in the cassettes themselves. Are you using mesh cassettes? Are you loading the trays with cassettes into an empty retort and then starting the delayed program? If so, air pockets could be forming in the cassette and around the tissue pieces. Since there is no p/v cycle running yet, those air pockets may be just sitting there and the exposed tissue pieces may be getting dried out. I suggest that once the retort is full and in ambient mode, open retort and move the cassette trays around a bit and see if air bubbles are rising from the cassettes. When the trays of cassettes sit on the bench in formalin waiting to go on the VIP, there may be enough movement when moving the container, etc, that any air pockets are disrupted and the tissues are not exposed to air pockets. good luck, amysue ruppert Marshfield Labs Histology From: histonet-requ...@lists.utsouthwestern.edu Sent: Monday, February 5, 2024 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [EXTERNAL] Histonet Digest, Vol 243, Issue 4 CAUTION: This email originated from outside of the Marshfield Clinic Health System. Do not click links or open attachments unless you recognize the sender and know the content is safe. Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit https://urldefense.com/v3/__http://lists.utsouthwestern.edu/mailman/listinfo/histonet__;!!J7HzeEKFbK9hUUY!PGATEy43_xJX9a3K3Na-c7VKlFfxZ5567gwny4UONDtiVRyrbBI5aHBtxx6V0DTFWbIgFABz11BpPHqIM6HdmZ9BO2ad6vok37eGCo6_gU_9gOLs_lTLCQ$ or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Processing artifact - delayed start (Bacon, Charles) -- Message: 1 Date: Mon, 5 Feb 2024 13:07:41 + From: "Bacon, Charles" To: Verizon wireless , "histonet@lists.utsouthwestern.edu" Subject: Re: [Histonet] Processing artifact - delayed start Message-ID: Content-Type: text/plain; charset="utf-8" We have had 2 reasons why we saw processing issues like this: 1. I recently found out on our VIP 5 they did not turn the level sensors on during install. These sensors are known to error so often, Sakura tells technicians to set the default to off. All the processor can sense is pressure and time. So it may be that the formalin is not filling all the way. You are noticing this on delayed runs, but on those runs are you often stacking trays? If so, isolate the cases in the upper trays and review. 2. We found a clogged line. This happens with the formalin lines if you don?t do the hot water flush often enough. This can be an even bigger issue if you recycle and re-buffer your formalin (we do not). Good luck! Chuck Bacon, HTL(ASCP)CM Supervisor Histology Baystate Medical Center -Original Message- From: Verizon wireless Sent: Friday, February 2, 2024 10:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Processing artifact - delayed start Dear Histonet Members, We have terrible processing artifact if tissue sits in the formalin-filled retort (at ambient temperature) for too long (more than 10-12 hours) before a delayed process starts. The longer the wait, the worse it looks. We have Tissue Tek VIP 5 processors, and we process luminal gastrointestinal biopsies exclusively. I've attached some photomicrographs of problem cases on the Histonet Images website (with the same topic title). This artifact typically affects a few specimens per day (~2% or less), even though everything is done on the same processor; it may affect all tissue portions in a cassette or only some of the tissue in that cassette. Some tissue portions may only have the artifact on one half with the other half looking perfectly fine. The techs sometimes note the tissue feeling "crunchy" at the time of embedding and / or at the time of microtomy. These tissues tend to suffer greater chatter artifact and have trouble sticking to the slides. The sections look just as bad on recuts as the originals. Re-processing does not seem to help at all. If the cassettes sit in formalin in a container outside of the processor for days before the processor is loaded (with subsequent immediate start), things look perfectly fine. When we have staff around to start the processor immediately upon loading cassettes and empty immediately upon completion, the tissue looks perfectly fine. Our current processing program is as follows: 1. 10% Formalin, 30 minutes, ambient temp, p/v on 2. 10% Formalin, 30 minutes, ambient temp,
[Histonet] Teaching color blind Histo student
Hello, we currently have a color blind Histology student. Does anyone have any helpful hints to share about learning in the Histology setting with colorblindness? Note, we had previous student with colorblindness, many years ago, and he did fine. But most of the teachers that helped him are now retired. Thank you amysue ruppert __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryostats decontaminationIn-Reply-To=
We use an Isopropanol based spray called DisCide Ultra Disinfecting Spray, ColePalmer catolog # EW-86306-12. Or direct from the company that makes it, www.palmerohealth.com. The spray bottle that it comes in puts out a very fine, wide spay of the disinfectant which gets it into all those hard to reach spots of a cryostat. We do this weekly along with the UV light decontamination that one of our cryotstat is equiped with. We do a more extensive/take down decontamination monthly. amysue ruppert __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] processing program for biopsiesIn-Reply-To=
I agree that the time may be too short. I would suggest removing the 80% alcohol station and adding another 100%, with the time of 25 minutes. If you have a VIP tissue processor, you may also want to change the mix to fast, with such short times in the stations, the processor does not have any time to do a pump in and pump out when set on slow. If set on fast, you should get at least one mix per station. This is helpful because it breaks any surface tension created around the tissue pieces and will allow for better penetration of the solutions. I would also suggest to put the heat on the paraffins to 58 C. Too much exposure to heat can cause poor, hazy staining. If you have had this program set for awhile, and had good luck before, but now are having issues, than you need to look into the quality of the solutions on the processor. And make sure the processor is set up correctly..meaning the correct solution is in the correct station. It can happen that someone set up the solut ions incorrectly, since we are all only human and mistakes happen. good luck AmySue Ruppert,HT(ASCP) MB(ASCP) Marshfield Labs, Histology Lab __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Drying oven tempsIn-Reply-To=
For HE slides, we use the Leica ST5020 autostainer. We have two ovens. The first oven is set for 40 C, 20 min. Then the slide rack is transferred to the second oven at 60 C, 15 min. WE have recently incorporated placing all of our slide racks for HE in front of a small table top fan for 10-15 minutes prior to going onto the autostainer. This combination works well for us and does a great job of cutting down/ eliminating on the amount of nuclear bubbling artifact. We are not so stringent with our special stain slides. And IHC slides also have their own protocol. amysue __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MethenamineIn-Reply-To=
It is also available through Fisher. Hexamethylenetetramine H290-500 for a 500 gram bottle. amysue __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thanks to Leica serviceIn-Reply-To=
One point to bring up, and I am sure that Leica service will do this, is the brand of knives you are using. Different brands have slightly different thickness. This is not related to the height of the blade. We have found that we need to use the blades that are used by our service company to calibrate the microtomes to have the correct tension in the holder and not any problems with the quality of the sections. good luck, seems you are on the right track though. amysue ruppert __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Leica Service TechnicianIn-Reply-To=
We moved into a new lab a few years ago, and due to the incoming air current with the ventilation here, it caused our cyrostats to ice up considerably. We have the type of ventilation that has air vents on the ceiling and are constantly blowing large amounts of air into our rooms. We had to get a piece of plastic put into the vent to divert some of the air away from our cryostat. It helped, but still ices more than when we were in our old lab. Anyway, point is to check out the air flow above the cyrostat. You also need to have enough clearance behind the cryostat and on each side. The clearance amount would be listed in the manual. good luck amysue __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] autoclave water
Do these jars go through your lab dishwashing dept? Could it be soap residue or some other residue from the dishwashing? Have you tried to rinse the jars well prior to putting the water for autoclaving? The dishwashing dept should be able to test for any residue. amysue __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE%3A Verhoeff/Masson%27s StainIn-Reply-To=9BF995BC0E47744E9673A4148
Hello John, Just reading through some histonet archives and found your Elastic- Masson issue. Some notes on the elastic masson stain The fibers should darken when the slides go into running water after the 2% ferric chloride. So the color before the ferric chloride will not be the same intense color as after the running water. But one very important point is that with this stain, you should actually UNDER differentiate the slides in the 2% ferric chloride because the solutions in the masson part of the procedure will also differeniate out the fibers. And so you could actually wipe out the fiber staining after going through the entire procedure. In our lab, the technique is to do only 2-3 dips in the 2% ferric chloride (yes, this will seem like not enough time in the ferric chloride, but remember, the masson solutions will also differentiate the slides) and then procede with the rest of the staining and then you should be seeing the fibers. Also, do not be afraid to try to contact PolyScientific, they are a very helpful company. Good luck. amysue ruppert __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Light hematoxylin staining in H%26EIn-Reply-To=
We seem to be constantly battling this as well. We have found that the quality of the ethanol used in processing and staining can be an issue. We now purchase ours from a company at www.UltraPure.com. The quality of the DI or distilled water being used can also be an issue. We are currently in the process of getting off of our lab building DI water, and purchasing a Elix Advantage Purification system from Millipore. We also found that the type of Eosin can make a big difference. We have recently changed to Eosin from Anatech, and this gives us a very good end product with our Hematoxylin that we still make from scratch--yes we still make our own:) However, the best results we get are when we do not use the DI water that comes from our buildings water system. That is why we are working hard to get our dept our own purification system. It is a very long story how we have come to this decision. But checking into your water supply and the quality of it would be very worthwhile. Very few people realize or understand the importance of water quality for the histology lab. Good luck amysue ruppert __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAS for basement membraneIn-Reply-To=
We have found out that we need to use freshly prepared 0.5% Periodic Acid. Your times seem appropriate. amysue ruppert marshfield labs, Histology. __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAS for basement membraneIn-Reply-To=
Yes, it is good to use the metabisulfite rinses to remove any excess schiff reagent after the schiff step. But after the metabisulfite rinses, you still need to go into TAP water for several minutes to have the full color of the PAS reaction develop. It will get much brighter after the final tap water rinse. amysue __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Poor nuclear artifactIn-Reply-To=
Are the individual pieces that have the staining problem larger than the rest of the pieces in the same block? What type of tissue is this happening with? Is it happening with tissues that are coming from one particular account? It seems that if the staining issue is not across the board with all your tissues or even with tissue pieces in the same block, than you need to look at some of the above questions. Tissue such as a skin tag may seem small enough for a bx run, but in reality, tissue like this is sort of like a basketball The tough outer skin does not allow for the processing solutions to pass through easily and penetrate as it should. Even a large skin tag or colon polyp that is bisected may have this happen on those pieces that are like half a basketball...solutions get trapped inside, but cannot penetrate pass the outer shell. And so these pieces end up improperly processed. Or, if this is happening with a particular account, could something have changed there..new pe rsonnel that may not know the importance of proper fixation, and are individual tissue pieces drying out before being placed into the fixative? You mentioned Alaska, could any of these possibly have become frozen or partially frozen in transport? good luck AmySue R. __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Amylase Digestion for glycogen
Hello, We are looking to switch from malt diastase digestion for glycogen to Amylase digestion. I have the new protocol worked up, but one of the Pathologists I work with would like to have an idea of how many labs out there are using Amylase instead of malt diastase for their PAS/D method. If you use amylase for the PAS/D method, could you please let me know who you are and the institiution? Much appreciated. amysue ruppert Histology lab Marshfield Labs Marshfield WI __ 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
