Re: [Histonet] Histonet Digest, Vol 208, Issue 14 Best Slides for Leica IP S Slide Printers

Be careful. Look at the slides before you commit. The least expensive may also be the dirtiest. So stack up 12 slides and look through them. If they look cloudy or dirty. Nix Do the drop test. A slide should be able to survive being dropped to the floor from waist high. Choose on price after

Re: [Histonet] Histonet Digest, Vol 207, Issue 3 Bouin's vs Vinegar

If the desire is to fix tissue marking dyes on tissue, 5 % acetic acid (vinegar) works well and does not stain or alter/fix the tissue like Bouin's fixative. Thanks, Steve Steve A. McClain, MD 631 361-4000 cell 631 926-3655 -Original Message- From: histonet-requ...@lists.utsouthwestern

Re: [Histonet] Histonet Digest, Vol 206, Issue 1 New Tissue Processor? (Sandra Cheasty)

While (conceptually) looking forward to the virtues of new and improved modern tissue processors w WiFi and SMS messaging capability, I fully expect to be operating the 1979 K-series for years to come. I reckon we may be behind the times, but with regular annual maintenance, our VIP5 and even

Re: [Histonet] Histonet Digest, Vol 201, Issue 3 Update info. on old topic?

Tim, Paula and Jeanine I have little experience w slide scanning, yet I have captured nearly 1.8M slide images, mostly on film coverslipped slides. One year ago we switched back to glass. 1) I suspect what Tim describes may be due to the fact that the refractive index of film differs from glass.

Re: [Histonet] Histonet Digest, Vol 197, Issue 23 recovering RNA from FFPE

1) Does anyone know of a method of recovering RNA from FFPE? 2) Does anyone know a lab that has done it? Thanks, Steve A. McClain, MD McClain Labs 45 Manor Road Smithtown, NY 11787 631 361-4000 cell 631 926-3655 ___ Histonet mailing list Histonet@lists.

Re: [Histonet] Histonet Digest, Vol 197, New Topic Coronavirus testing from tissue sample

The CDC website with specimen submission form does not seem to be working. https://www.cdc.gov/laboratory/specimen-submission/pdf/form-50-34.pdf Does anyone know who can else can test for Corona virus from FFPE tissue? I have an unusual skin biopsy from late December (earlier than the first known

Re: [Histonet] Histonet Digest, Vol 196, Issue 3 Incomplete sectioning

Two methods to avoid incomplete sectioning are: 1) ink the specimens well, using acetic acid (vinegar) to fix the ink, thereby making the ink easier to see in the block 2) have a microscope or 10 x lens adjacent to the microtome to allow for visual inspection. One of my histotechs used an old (r

Re: [Histonet] Histonet Digest, Vol 194, Issue 12btattoo removal

I don not know any method for tattoo pigment removal. Aside from carbon black, most of the red, yellow, blue and green pigments are metal salts and polarize brightly. The inflammatory response may be vigorous, especially w repeat or Re-do of a tattoo. In some cases a pseudo-carcinoma or KA may

Re: [Histonet] Histonet Digest, Vol 189, Issue 26 Tissue Contamination /Floaters

I wrote and posted on this subject some years ago. In our lab, we virtually eliminated floaters 10 years ago, by using a combination of behavioral and technical procedures: 1) photographing every specimen since 2004 (in order to achieve report -worthy gross images, one must keep the grossing bac

Re: [Histonet] Histonet Digest, Vol 189, Issue 13 PRAME antibody for melanoma

Would one of the LIST members kindly share their experience with PRAM antibody in paraffin/ Steve A. McClain, MD * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailm

Re: [Histonet] Histonet Digest, Vol 188, Glass plate for AO knife sharpener

Does anyone know where to find the glass plate for an ancient AO sharpener for microtome? Steve Steve A. McClain, MD 631-361-4000 Cell 631-926-3655 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/list

Re: [Histonet] Cassette labeler

Anyone care to recommend their favorite cassette labeler to replace the Microwriter? Thanks Steve Steve A. McClain, MD 631-361-4000 Cell 631-926-3655 On Jun 24, 2019, at 13:19, "histonet-requ...@lists.utsouthwestern.edu" mailto:histonet-requ.

Re: [Histonet] Histonet Digest, Vol 179, Issue 4 AFB control

I was taught it best to use separate controls on a second slide. But in practice, the only cases where a floater appeared it was obvious. In my own lab 14 year experience it has not happened. However, about 10 years ago we started an experiment and switched to 2 AFB slides with pos and neg cont

Re: [Histonet] Histonet Digest, Vol 170, Issue 13 histology hacks

I purchased the book and applaud the effort because there is some decent information. However, the term hack is a poor choice for histology and many of the fixes or secrets described are because some hack failed to do her/his job at an earlier step in the process. (Definition of hack. transitiv

Re: [Histonet] Histonet Digest, Vol 168, Issue 2 Needing a microwave processor

Why do you think you need a microwave processor? You can certainly use a conventional K3000 or VIP5 or RVG (and probably other) processors to accomplish short 2 hours-4 hours processing on small samples by 1. Raising the processing temperatures to 45-50C 2. Removing formalin from the processor (al

Re: [Histonet] Histonet Digest, Vol 164, Issue Olympus microscope camera DP70 works on Windows 10

Yes, to my amazed relief, Olympus microscope camera DP70 works on Windows 10. These old but reliable cameras can be upgraded from Windows 7 (Vista drivers) to Windows 10. We just upgraded one of my workstations and it works fine. Steve A. McClain, MD

Re: [Histonet] Histonet Digest, Vol 164, Issue 10 mycoplasma on agar

Too bad, Hoescht stain is rapid and sensitive (UV) for mycoplasma. They can try staining direct smears w difquik giemsa. Or fix agar plate in Carnoy's and slice out and process to paraffin for microtomy and staining. IHC can be done from sections if needed. These 1 micron bacteria are at the

Re: [Histonet] Histonet Digest, Vol 164, Issue 9 slide adhesion

Greg, It sounds like 2 or 3 problems and LEEP specimens are partially heat fixed, giving addition artifacts related to poor adhesion. 1. Short processing cycles w insufficient paraffin times or dirty xylenes can do this. Double your maintenance or try the radical by removing formalin from proc

Re: [Histonet] Original 1979 sales price for Miles Tissue Tek 3000

Does Dr. Richmond or anyone else recall the original sales price for the Miles TissueTek VIP 3000? Some of these vacuum processors are still in operation almost 40 years later! Steve A. McClain, MD ___ Histonet mailing list Histonet@lists.utsouthweste

Re: [Histonet] Histonet Digest, Vol 163, Issue 22 CPT Code for PGP9.5 free floating tissue stain (Eddie Martin)

Depends also on what you are doing w the pgp9.5 The only reason I know for free floating section technique is for intraepidermal nerve fiber analysis. Morphometric analysis and nerve fiber counting is an additional CPT. Steve A. McClain, MD > On Jun 24, 2017, at 13:09, "histonet-requ...@lists.u

Re: [Histonet] Histonet Digest, Vol 163, Issue 15 Research histology and animal IHC

Underfixation being the greater problem, for animal research I tell everyone 24 hours fixation. After 24 hours in clean formalin, Stop fixation by pouring off and transfer to 50% reagent alcohol. Using standard fixation and slow processing, repeatable near-perfect histology and IHC can be made w

[Histonet] (no subject)

If it is any help, A dollar bill weighs 1 gram and a quarter weighs 5.67 grams. It is simpler to write up some calibration procedure based on knowns. Steve A. McClain, MD > On Jun 8, 2017, at 13:20, "histonet-requ...@lists.utsouthwestern.edu" > wrote: > > Balance calibration > Message-ID: _

Re: [Histonet] Histonet Digest, Vol 162, Issue 26 butterflies

I have no experiences w butterflies, but the chitin in ticks, carpenter ants and spiders does not cut well. For insects and arthropods we post-fix in Carnoy's for 60 minutes Then soften with 10% KOH RT for 10-20 minutes then wash for 30-60minutes in water. Long processing cycle. Steve A. McCl

[Histonet] Blades

http://www.emsdiasum.com/microscopy/products/preparation/blades.aspx Shared via the Google app Try electron microscopy sciences. They have a variety of blades. Steve A. McClain, MD ___ Histonet mailing l

[Histonet] PCR for tick bite vasculitis

Does anyone have experience or suggestions for where to send FFPE biopsy sections from a patient with erythema migrans and known Amblyomma tickbite. Dieterle negative, anti-Borrelia burgdorferii negative Gram negative PAS negative. Thanks for your help. Steve A McClain, MD 631-361-4000 ___

Re: [Histonet] Histonet Digest, Vol 162, Issue 6

Olympus microscopes win in my experience. You can't go wrong with any of the 3 major manufacturers. Modern scopes all have have better optics than what we trained on. I place a high priority on durability and in my experience with Nikon and Olympus, Olympus has consistently shown better overa

[Histonet] Fwd: Histonet Digest, Vol 162, Issue 6 micrcroscopes

Olympus microscopes win in my experience. You can't go wrong with any of the 3 major manufacturers. Modern scopes all have have better optics than what we trained on. I place a high priority on durability and in my experience with Nikon and Olympus, Olympus has consistently shown better overal

Re: [Histonet] Histonet Digest, Vol 161, Issue 24 ICD10

1 Google it or look it up Or 2 use an allpurpose icd10 as a placeholder, e.g. For skin one can use r23.8 other skin changes, unless you can find an icd10 for "no icd10". 3 feeling bold- use V91.07XA (burn due to water skis) or V96.00XS, which outlines an unspecified balloon accident injuring occ

Re: [Histonet] Histonet Digest, Vol 161, Issue 5Subject: Re: Processing cystic tissue

Skin cysts are difficult for everyone. Cysts and toenails lead the top of our list of cases with catastrophic tissue loss - which we define as tissue loss w "holes" >1mm2 square. The cyst center generally does not fix readily, composed of unfixed cornified cells and waxy lipid. On the scalp, w

Re: [Histonet] Histonet Digest, Vol 160, Issue 30 Subject: disposable grossing pad for specimen imaging

We use squares of absorbent paper towels to first blot excess fixative and then place on a 2mm thickness piece of art foam. Art foam is used as a background for photography and slicing. By sliding or moving the foam we position the specimen under the camera during specimen photography. We have p

Re: [Histonet] Histonet Digest, Vol 159, Issue 20Subject: simple tests for xylene contamination and alcohol conc. in Recycled alcohol

at 13:22, Steve McClain wrote: > > We only use recycled reagents (except 100%) on processor. > We only use Fresh reagents on stainer and coverslipper. > Steve A. McClain, MD ___ Histonet mailing list Histonet@lists.utsouthw

Re: [Histonet] Histonet Digest, Vol 159, Issue 20Subject: Recycled reagents in VIP processor

We only use recycled reagents (except 100%) on processor. We only use Fresh reagents on stainer and coverslipper. Steve A. McClain, MD ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Histonet Digest, Vol 159, Issue 16 Colloidal iron leaching

After checking reagents and glassware, look for other sources of iron contamination, e.g. Metal forceps, steel staining racks, water bath. Inadequate rinsing after the colloidal iron step - increase wash times, using DI Check concentration of working reagent. Decrease the time in colloidal iron

Re: [Histonet] Histonet Digest, Vol 158, Issue 22 Breast Specimen

In my experience, rushing to process fatty or inadequately fixed specimens is a fool's game. In my opinion, this problem cannot be solved by the histotechs- it begins with the grossers and is one for the pathologists to solve at the grossing bench. Suggestion #1 Do nothing and let the medical

Re: [Histonet] Histonet Digest, Vol 157, Issue 21 the center

What paraffin are you using? I heard a rumor one manufacturer changed formulations recently. Same brand name different formula. Center problems often manifest where insufficient fixation is a primary event or where rapid processing meets the limits of solution maintainance and tissue size. R

Re: [Histonet] Rapid processing With Conventional Processors and conventional (old school) reagents

min Station 9 Xylene 10 min Station 10 Xylene 10 Min Station 11 Paraffin 5 min Station 12 Paraffin 10 min Station 13 Paraffin 10 Min (VIP5 Station 14 Paraffin 10min) Total Time to complete ~2:20hours Steve A. McClain, MD > On Dec 9, 2016, at 17:37, Steve McClain wrote: > > Station 1 5

[Histonet] Rapid processing With Conventional Processors and conventional (old school) reagents

I posted this previously. We have done this more than 2400 times with a number of processors. It can be done if you pay close attention to processing only FIXED tissues, use fresh reagents and lay out the cassettes not more than two high. Steve 631 361 4000 Station 1 50% 10 min Station 2 70% 10

Re: [Histonet] Histonet Digest, Vol 157, Issue 5 outdated gold

Cheers! We were dinged on the last inspection for having outdated reagents near the human testing. We have a gold standard in the lab, specifically a Queen Victoria 0.25 ounce gold coin minted in 1870. It appears to be past expiration, yet still attractive and untarnished. Any suggestions fo

Re: [Histonet] Histonet Digest, Vol 157, Issue 1Glypican-3

Victoria Verno You might try normal Placenta or fetal liver from POC as non-neoplastic stain controls. That strategy may allow you enough repetitions to develop the stain until you find a case of HCC to test. Steve A. McClain, MD * ___ His

Re: [Histonet] Histonet Digest, Vol 156, Issue 20a hematoxylin-eosin Y-Azure-II stain

hematoxylin-eosin Y-Azure-II stain Agree w Ren. But try it anyway. To bring out the azure, you may wish to post fix. Steve A. McClain, MD > On Nov 22, 2016, at 13:29, "histonet-requ...@lists.utsouthwestern.edu" > wrote: > > a hematoxylin-eosin Y-Azure-II stain _

Re: [Histonet] Histonet Digest, Vol 154, Issue 13 tissue loss

Wendy Develop your own data. Check your early steps first. Tissue loss can be simple -a function of poor fixation, dirty processing or rush processing or slightly large tissue w slightly dirty reagents. Airdry wet slides before annealing on hot plate. Check your baking/annealing. Some tissues r

Re: [Histonet] Histonet Digest, Vol 154, Issue 7ubject: fixing fatty skin specimens

Not sure I understand your question. But assume you are looking for margins on a re-excision of cancer or melanoma. Are you using a sufficient -20 times volume of clean 10% nbf? And changing it often? Switch to 20% nbf Or switch to 100% formaldehyde! True dat- Stanford's Dr. McNeal fixed prostat

Re: [Histonet] Histonet Digest, Vol 154, Issue 2 movats

Liz, please send SOP ASAP! The vvg is a hematoxylin-that's why it works so well. The problem w Movat is the photography. It is complex w 5 color signals, hard to show all 5 in one image. Liz, What control did you use? Steve A. McClain, MD > On Sep 2, 2016, at 13:13, "histonet-requ...@lists.utso

Re: [Histonet] Histonet Digest, Vol 153, Issue 24 davidsons fixative fish

Something sounds fishy Dr. Richmond can solve it. http://www.ihcworld.com/_protocols/histology/davidson_fixative.htm Steve A. McClain, MD On Aug 30, 2016, at 13:22, "histonet-requ...@lists.utsouthwestern.edu" mailto:histonet-requ...@lists.utsou

Re: [Histonet] Histonet Digest, Vol 153, Issue 10

For once I almost refuse to answer because there is No good answer. This is absolutely ludicrous. Heck no is my first response but that probably doesn't pass muster. My state inspectors are probably wanting to have me fill out more firms forms for monthly bleaching. Or adding a bleach step to

Re: [Histonet] Histonet Digest, Vol 153, Issue 3

> Today's Topics: > > 1. Wax clogs in VIP 6 processor (cynthia haynes) > 2. Re: Histonet Digest, Vol 153, Issue 2 microtomes (Steve McClain) > 3. slides on the Benchmark Ultra (Amy Johnson) > > > -

Re: [Histonet] Histonet Digest, Vol 153, Issue 2 microtomes

Depends on how long you plan to be in business and whether you are using other people's money. But buy all the same model and make if you can. I've owned the same Leica 2125 and 2135 microtomes for 12 years. We usually cut mainly small biopsies but have pushed those to occasionally cut large

Re: [Histonet] Histonet Digest, Vol 153, Issue 1

I agree w Dr. Richmond. > > Message: 2 > Date: Sun, 31 Jul 2016 17:43:20 -0400 > From: Bob Richmond > To: "Histonet@lists.utsouthwestern.edu" > > Subject: Re: [Histonet] B-5 Fixative > Message-ID: > > Content-Type: text/plain; charset=UTF-8 > > Jillian A. Russell, HT (ASCP)CM, QIHCCM, S

Re: [Histonet] Histonet Digest, Vol 152, Issue 18

PAS crystals. The problem may not be the stain at all. occurring due to conditions to leaching of biochemical substances, in the tissue. Dirty processors, or use of KOH or acids applied to the block just before sectioning. Those may not be washed out. These crystals are especially common in

[Histonet] 360 video of specimens

The next 3 weeks we are turning our lab cameras and our specimens toward making a 360 photographic orbits around specimens to create a 360 stitched image or video. If you know any pathologists who have tried 360 imaging around a specimen, kindly send me their name. I have not seen any prior art

Re: [Histonet] Histonet Digest, Vol 152, Issue 9 lab startup

This is a challenging topic. We did it in 2004. You must have deep pockets and committed clients. Even then, it is a hunter-gatherer lifestyle. Run your traps, and live off what you kill. It can be done, but the wind is always in your face and the first 7 years are mostly uphill. Good luck S

Re: [Histonet] Histonet Digest, Vol 150, Issue 31 5. Re: those slippery lab floors (Normington Lacy)

I did see many slip and fall injuries in histolabs over the years. When we built this lab we accounted for that and deliberately left the bare concrete rough, and professionally painted polyurethane of the type used in firehouse floors. Has held up reasonably well. No slips or falls in 12 year

Re: [Histonet] Histonet Digest, Vol 150, Issue 21 floaters

Never seen that happen in 34 years of training and practice. Sounds like an inadvertent transfer by the grosser. But Floaters are so frequent that it behooves labs processing small biopsies to wrap each specimen to minimize or eliminate transfers. We have the embeddets clean the forceps w th

Re: [Histonet] Histonet Digest, Vol 149, Issue 7 : Re: specimen submission pads ordinary lens paper

There is much commendable about ordinary lens paper cut from 9x12 sheets to ~2x2 inch and folded 4 times like an origami to enclose small tissue fragments. Keeps floaters out. Translucent so you can count the fragments before and after opening. Unfolds reliably at embedding wo flinging pieces. C

Re: [Histonet] Histonet Digest, Vol 148, Issue 20 processing

I don't really like wasting valuable rapid-processing time on fixation, when I cab icontrol that at the bench Your schedule is made more difficult because of the length of time in formalin. See if your director can ensure only fixed tissue on processor. If not increase temp to 45C station #1 an

Re: [Histonet] Histonet Digest, Vol 146, Issue 10 tissue processors

Sakura VIP you can run for decades if maintained. Extremely reliable. The parts are available. I have 3 K series and a VIP5. The newest is 17 years old, the oldest maybe 35 years. All acquired used and refurbished. Our newest processor, 7 years, a demo acquired almost new, failed miserably t

Re: [Histonet] Histonet Digest, Vol 146, Issue 4 validation suggestions for short cycle processing with conventional equipment

I posted our 7 year experience on this topic previously on histonet. Remember, this machine (K3000, VIP5) was not designed specifically for this purpose, but it can be made to work admirably. IOW Short processing on VIP is unreliable UNLESS you are willing to accept or control certain conditi

Re: [Histonet] Histonet Digest, Vol 145, Issue 15. 6. Solvent recycler (Rachel)

BR for 11 years. We recycle alcohol and xylene for reuse on processors. Extremely reliable. Steve A. McClain, MD > On Dec 16, 2015, at 13:11, "histonet-requ...@lists.utsouthwestern.edu" > wrote: > > 6. Solvent recycler (Rachel) ___ Histonet mailing

[Histonet] Miniature arrays for control

We have used miniature arrays for 9 years. We place both positive and negative controls on every IHC. Minimal design is one positive and one negative control. To start make a simple one with positive and negative using a 2mm skin punch biopsy with plunger to cut out known tissue and mount toget

Re: [Histonet] Histonet Digest, Vol 145, Issue 6 Oct separating from epidermis

Skin is often difficult to cut. Not sure I have seen that artifact, but here is what I use when having difficulties. Gently blot the tissue/epidermis dry before immersing in liquid OCT/Neg50. Then freeze in cryostat. Avoid refrigerant spray. Sometimes we allow tissue to sit in oCT for 5 min and

Re: [Histonet] Histonet Digest, Vol 144, Issue 21: Rapid tissue Programs

Since processing is not the issue, first standardize fixation, eg 4-6 hours before processing or in the first reagent on processor. Change that first formalin every 200 blocks or so. When well-fixed, small tissues will tolerate rapid processing. Finally, if your pathologists can read the slides

Re: [Histonet] Histonet Digest, Vol 141, Issue 5 unexpected AP findings

We have pathologists call nearly all unexpected findings. Experience has shown that some findings we initially viewed as "insignificant" later proved to be problematic. We document calls to and from physicians (slide reviews also) in the reporting database record and usually in the report with

Re: [Histonet] Histonet Digest, Vol 141, Issue 4Surgical Pathologist Benchmark

The best benchmark is the one you derive with your personnel and your pathologists in your system. The benchmark number you seek is not readily available because work is primarily dependent on the efficiency of your system of reporting, whether the pathologist has to spend time proofreading th

[Histonet] Tissue processing validation (Ann Specian)

This seems like overkill. Just moving a processor previously validated should not require re-validation, just verify the programming and make certain the paraffins are melted completely. This may not apply to all processors, but for all Sakura machines manufactured since 1980,I would not revali

[Histonet] Decalcification and Neutralization (Huynh,Thomas)

I have not found the need for neutralization. With sufficient washing, common histochemical stains including H&E Gram PAS-fungus and trichrome are spectacular. Please explain your protocol for neutralization. Steve A. McClain, MD ___ Histonet mailing l

[Histonet] Decalcification

One frequently overlooked point is washing after decal. Especially important with acid decal, we wash for an hour in running water. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Manual photography and Picky Pathologistsl

Picky pathologists only want the best. They are a necessary evil. The problem is defining objective problems as opposed the perceived colors. To simplify the discussion, assume for the moment your pathologist has been evaluated and is NOT colorblind. To take the emotions and subjectivity out o

Re: [Histonet] Histonet Digest, Vol 140, Issue 6 Specimen loss

ecialists.com] Sent: Wednesday, July 08, 2015 2:40 PM To: Steve McClain; histonet@lists.utsouthwestern.edu Subject: RE: Histonet Digest, Vol 140, Issue 6 Specimen loss Dear Mr. Cantankerous, :) So how would you gross a colon biopsy that was <0.1 cm and probably mucus? -Original Message--

Re: [Histonet] Histonet Digest, Vol 140, Issue 6 Specimen loss

My goodness 7% tissue loss is ridiculous. What fool published that? A 0.8% "loss of tissue" is beyond me. Would any one advertise in the local paper that your center of excellence lab loses nearly 1% of your specimens? "possibly too small to survive processing" is in my opinion, a self-fulfillin

Re: [Histonet] Histonet Digest, Vol 140, Issue 5 specimens lost

Steven, The problems associated with tissues lost are quite similar to those with floaters. Here is a newly edited copy of what I posted a few years ago on floaters and you may find it useful. We still wrap nearly all specimens. At times we cannot see minute flakes of tissue, but retrieve them

[Histonet] RE: Histonet Digest, Vol 128, Issue 29

I requested 15 levels at 200 microns, that gives us 3mm of detail 5 slices per mm. Steve 631 361 4000 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Tues

[Histonet] Grossing without gloves

If you look back 100 years to when gloves were first used in medicine, you will find that gloves were initially used in surgery to protect hands of surgeons and nurse-assistants from caustic effects of carbolic acid and not directly to prevent patient infection. Grossing without gloves is but

[Histonet] RE: Histonet Digest, Vol 112, Issue 34 genta stain

May well work-Genta stain has AlcianBlue. Please note HL Steedman's original paper describes "formaldehyde fixative is not good" for stabilizing/fixing mucin. Steve A. McClain, MD 631 361 4000 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu

[Histonet] VIP6 issue

I may be oversimplifying, but I see that as one of many reasons for controlling fixation on grossing bench, Then rinsing in alcohol and starting in alcohol on the processor. 50% Alcohol has less surface tension than water and fewer air bubbles. Blue sponges have a number of annoying artifacts inc

[Histonet] eosin on processor

Eosin on processor can be done with dirty eosin waste saved from the stainer; adding eosin powder sounds like a mess. Especially helpful for orienting small samples with a sidedness, skin GI GU oral, cornea No common down sides for routine HE, PAS, Trichrome, or IHC even with Red chromagens. We

[Histonet] 88305

Looks like rough seas ahead. Raising my research fees 33%. Investing to start a new patient center to do more procedures. After that we may have to get creative. Steve Steve A. McClain, MD McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000

[Histonet] holes in derm tissue sections

Generalized Holes commonly present with one of two scenarios. Scenario One-a Histotech may be sloppy about allowing sections to dry thoroughly before warming or baking. Scenario Two is generalized, showing up with slides from every Histotech, and representing unstabilized foci where paraffin di

[Histonet] RE: Histonet Digest, Vol 108, Issue 19 H&E staining

Eosin tissue affinity vs hema is far different with staining times measured in seconds for Eosin vs minutes for Hema. Overstained Dark eosin makes H appear too light by contrast. If one considers an analogy of a balance or seesaw with H on one end and E on the other, your solution is obvious. Whe

[Histonet] Inking skins

Many apologies for being cantankerous, BUT NEVER INK TISSUE WITH A COTTON TIPPED APPLICATOR Just to be clear- NEVER Cotton fibers stick to tissue and remain in block. Fibers hang on microtome blade during cutting, causing tears and artifacts in final section. I've heard it doesn't matter, we do

[Histonet] Counter stain for PAS for fungus (Steve McClain)

Change all your reagents. Change periodic acid every week at least. PAS unlike many other stains is a chemical reaction and is sensitive to oxidizer Personally I see little advantage in fast green, and a common downside is overstaining in many labs, hiding fungus and basement membrane. Faint He

[Histonet] counter stain for PAS for fungus

Change all your reagents. Change periodic acid every week at least. Personally I see little advantage in fast green, and a common downside is overstaining in many labs, hiding fungus and basement membrane. Faint Hematoxylin is acceptable counterstain. For certain applications, e.g., dual staining

[Histonet] processor solution rotation

May I suggest? Define a standard that works for you, combining both elements, blocks and runs e.g., Every week or 200 blocks then check the first reagent volume and quality. Again at 400 And again at 600 blocks. We change the first reagent whenever it appears dirty. Consider adding a few ccs. of

[Histonet] Avidin Red for Mast Cells fading

Avidin Red or Avidin bound to Texas Red produces a beautiful specific stain for mast cell granule heparin. Triple stain (RGB) IF can be done by combining with fluoroscein labeled ab and a Hoescht stain for nuclei. However, the Avidin staining dissipates within days to weeks. Any suggestions?

[Histonet] RE: Histonet Digest, Vol 97, Issue 5 Cassette Marking Slide labeling

Leica sells the Surgipath brand cat. #3801880 a decent felt-type solvent resistant marker my techs like. (still have and like hand written slides and techs initials). Those black Tissue-Tek histo pencils are good too if you write tall, but for slides can carry some graphite dust onto the tissue s

[Histonet] RE: Histonet Digest, Vol 96, Issue 33 xylene and ethanol recycling in lab

Rene, I am interested to learn more of your experience with isopropanol as a clearing agent. Does it take longer to clear than xylene? For processing how many stations and how long are the times you use for clearing? How often do you maintain or change isopropanol - how many blocks per liter? W

[Histonet] RE: Histonet Digest, Vol 96, Issue 24 reprocessing

I just talked to Joel who reminded me. Mouse livers are often hard/ brittle, even under the best of circumstances. Avoid heat when processing, Room temp only Gradual dehydration 50/50/70/95% alc. 1 Hour per station. Steve 631 361 4000 ___ Histonet mail

[Histonet] RE: Histonet Digest, Vol 96, Issue 24 reprocessing Other methods

Not appropriate for your current mamafufu, But here are two 're-processing' methods used in our lab for small fixed tissues. Method 1 where the tissue was bit too large for a short processing cycle and the center of the block is sub-optimally infiltrated or soft and needs more paraffin time. C

[Histonet] RE: Histonet Digest, Vol 96, Issue 24 reprocessing

This sounds like a complicated problem- Generally fixed tissues left in the processor overnight can just be re-hydrated in formalin for a few hours and then run (we've made that mistake) Exceptions abound in histology methods, but Unless the tissue was well-fixed to begin with, Re-processing ge

[Histonet] RE: Histonet Digest, Vol 95, Issue 16 Drying Oven temps

It is important to clarify the terminology-what you mean by 'drying' oven? The remarks below refer to the manual step of annealing unstained sections on slides prior to de-paraffinization and staining. The point or objection to the terminology of 'drying' oven relates to a far too common practi

RE: Histonet Digest, Vol 95, Issue 19 [Histonet] Tissue processor

Mike, If your budget is tight, consider used Sakura VIP processor and Tissue-Tek embedding center. One superb vendor is Avantik , formerly Bellair in NJ. 800-783-9424 973-912-8900 They have earned my complete trust-I am reluctant to have equipment in my lab unless Bellair can maintain it. Every p

[Histonet] Histonet Digest, Vol 94, Issue 40 Nothing down the drain

Yes nothing down the drain (except maybe the lithium carbonate bluing). Hematoxylin goes into medical waste service. Used Eosin gets reused a 10-20 ml at a time into the first absolute on the processors as a marker of carryover (as the later alcohols turn red, we have a visual indicator of the ne

[Histonet] RE: Histonet Digest, Vol 94, Issue 38 Xylene safe disposal vs. aliphatic hydrocarbons e.g., Slide Bright, down the drain?

The argument against using toxic, yet recyclable xylene in favor of a more expensive, less efficacious xylene-substitute like Slide Bright in larger labs is not compelling. Aside from low volatility of Slide-Bright, what is gained in using a more expensive substitute whose toxicities are not well-

[Histonet] RE: Histonet Digest, Vol 94, Issue 35 XYLENE

It seems there is a debate on three or more issues, one being the validation of a test where xylene was the processing standard another being the safety of xylene where an employee is allergic or sensitized to xylene, a third on poor lab design and poor ventilation, another on the safety of xyl

[Histonet] Tissue tracking at embedding RE: Histonet Digest, Vol 94, Issue 24

Tim You may consider asking Dr. Phil LeBoit at UCSF Dermpath and see what he says. I wrote about floaters a few weeks ago and this subject is related in my view. McClain Labs uses a combination of 1) manual line drawings on the papers reqs to indicate # of pieces and approximate size; a.

[Histonet] RE: Histonet Digest, Vol 94, Issue 20 Question of combining immunofluorescence and more

Dual Fluorescence may give you a start. You can try Hoescht stain for nuclei and use another wavelength for the second signal. Eosin fluoresces , except eosin does not stain nuclei and being related to fluoroscein eosin works on the green wavelength. Steve Steve A. McClain, MD McClain Labs, LLC 4

[Histonet] Almost Eliminating Floaters

MY APOLOGIES FOR NOT DELETING THE REST OF THE HISTONET EMAIL FROM MY PRIOR RESPONSE. I HAVE ADDED ADDITIONAL NOTES AT THE END.- STEVE Floaters on the water bath are generally not the problem, are usually identifiable, unlike the serious issue posed by floaters in the block. I would suggest counti

[Histonet] RE: Almost Eliminating Floaters

Floaters on the water bath are generally not the problem, are usually identifiable, unlike the serious issue posed by floaters in the block. I would suggest counting to get a benchmark. In order to change the outcome, you (the institutional you) must change what you are doing. SO change your gro

[Histonet] Recycler (Behnaz Sohrab) xylene test for recycled alcohols product or alcohols to be recycled

7 years with the same B/R Procycler Would definitely buy again. It has paid for itself, even in a small lab. Good company. They will analyze your product and suggest revisions in the recycling programs as needed. How you use it and what you put into it to distill generally determines your r

[Histonet] gout procedure

Sandra, Gout is readily identified from Direct smears by examining either unstained, mounted slides or Dif Quik-stained slides mounted under polarized light. If processed, start in 70% alcohol and process to paraffin. Do not let slides sit in water during staining. Some uric acid gene

[Histonet] processing times for skin specimens

90 minutes for up to 2mm thickness using K3000 or VIP5. I do not have a VIP6 Steve A. McClain, MD McClain Laboratories, LLC 45 Manor Road Smithtown, NY 11787 631 361-4000 631 361-4037fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@list

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