Re: [Histonet] tissue cassettes

[patpxs via Histonet]

I'm old, old school too.  I prefer the paraffin pool myself, it helps prevent 
the bon-bon effect.  I had a vendor rep tell me the holding bins were designed 
for molten paraffin.Probably what happened over the years is that if there's 
too much molten wax in the bin, it over flows when the basket of cassettes is 
put in.Ah, displacement, just like when you get into an over full bathtub, 
water (or wax) everywhere. Now a days I think the dry holding tank is the rage. 
 As long as the tissue stays warm, it probably doesn't matter too much. Ciao 
bellas,PaulaSent from Samsung tablet
 Original message From: "Kurth, Virginia L via Histonet" 
 Date: 2/9/24  8:04 AM  (GMT-08:00) To: 
Thomas Podawiltz , "Brazie, Jeneanne E *HS" 
, histonet@lists.utsouthwestern.edu Subject: Re: 
[Histonet] tissue cassettes I am old school and prefer them dry, lol.  I agree 
with Thomas, that shouldn't have that affect.Ginny-Original 
Message-From: Thomas Podawiltz via Histonet 
Sent: Friday, February 9, 2024 8:34 AMTo: 
Brazie, Jeneanne E *HS ; 
histonet@lists.utsouthwestern.eduSubject: Re: [Histonet] tissue 
cassettesWARNING: This email appears to have originated outside of the UW 
Health email system.DO NOT CLICK on links or attachments unless you recognize 
the sender and know the content is safe.Without seeing the blocks, that sounds 
more like cold molds being used, more Then, whether or not the tissues are kept 
in a dry, hot, well, or a wet well.Sent from Yahoo Mail for iPadOn Friday, 
February 9, 2024, 6:00 AM, Brazie, Jeneanne E *HS via Histonet 
 wrote:Hello :) I am encountering push back 
in our lab when I fill the embedding units with melted paraffin in the 
embedding wells. The techs here like for the tissue cassettes  to sit dry (no 
wax) while in the  embedding units. I find that the tissue rolls out of the 
sections while cutting because of a layering effect between the tissue and the 
paraffin its embedded in. I have communicated this but they tell me I'm "old 
school". Does anyone have any thoughts or opinions on this 
topic??___Histonet mailing 
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Re: [Histonet] tissue cassettes

[patpxs via Histonet]

Hi Carl,The bon-bon effect.  This can happen if the wax used for processing is 
different from the wax used for infiltration or when the tissue gets cold 
before embedding.Either way, because the tissue is cold - the embedding wax 
doesn't blend with the tissue wax, creating the hard outer wax shell.  The 
simplest way to avoid the bon-bon effect is to make sure the tissue is as warm 
as the wax it's being embedded in.  Letting the cassettes go cold isn't the 
issue here, it's not letting the tissue warm up before embedding that is.  I 
call it "freezing" the tissue/cassette, since that's what the wax is doing, 
just to mess with people's heads.  That helps remind people that the wax gets 
cold quickly.Some folks may disagree with me about that.If the embedding wax is 
a different from the infiltration wax it's kind of the same thing.  The waxes 
need to co-mingle a little while so they can blend together before freezing the 
block.Just a few ideas...PaulaSent from Samsung tablet
 Original message From: Carl Hobbs via Histonet 
 Date: 2/9/24  11:12 AM  (GMT-08:00) To: 
histonet  Subject: Re: [Histonet] tissue 
cassettes I'm interested but don't understand the variation and it's + effectI 
take my cassettes out of the processor and immediately place into the molten 
wax bath of the embedder ( if I'm embedding immediately; if not I let the 
cassettes/tissues therein go cold until a later embedding)When embedding 
immediately, after 30 mins, I remove tissue from cassette into a heated metal 
mold filled with molten waxAll done quickly, to keep all wax moltenIf my people 
take too long, the wax around the tissue sets so when the tissue is placed into 
the molten wax, in mold, they sometimes get that effect of set wax-interface 
artefactthe section immediately pulls apart from the surrounding wax, on 
the waterbathThis can cause the section to foldDo enlighten meThanksCarlNever 
too old to learn!Carl Hobbs FIBMSHistology and Imaging ManagerWolfson SPaRCGuys 
Campus, London BridgeKings College LondonLondonSE1 1UL020 7848 
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Re: [Histonet] Automated Coverslippers

[Patpxs via Histonet]

Happy Friday Eve,

We use the Sakura glas2 cover slipper.  We run hundreds of slides through them 
every day with very few problems. 

Any automated coverslipper can have issues.  A word of warning- watching the g2 
do its thing is hypnotic.  

Paula

Sent from my iPhone

> On May 13, 2021, at 6:18 AM, Jennifer Phinney via Histonet 
>  wrote:
> 
> Hello Histonetters,
> My lab currently has a Leica CV5030 that is attached to our ST5010 
> Autostainer. We bought it way back in 2012 and it is starting to have 
> progressively more issues working reliably.  This is the only one that can be 
> integrated with our stainer, so we know we'll have to get a standalone unit 
> and transfer slides to a new rack.
> 
> What automated coverslipper is everyone using? I know in the past there have 
> been issues with tape coverslippers popping off of the slides when stored 
> long term, but has this been fixed with newer units? What is everyone's 
> experience with them?
> 
> I got a quote for the Leica Spectra coverslipper, but I am not familiar with 
> what other companies have available.
> 
> Thanks all,
> Jennifer Phinney  MS QIHC
> Histology Laboratory Administrator
> Kansas State Veterinary Diagnostic Lab
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Re: [Histonet] Tape Transfer Methods For Cryosectioned Brains

[Patpxs via Histonet]

Hi Heather,

I can see why you’re having trouble. 30 micron sections are inherently 
unstable.  Like paraffin, the thicker a section is the more difficult it is to 
cut.  Plus since your practice samples are old they tend to be more brittle. 

Try cutting at 5 microns and see what happens. Remember to let the samples warm 
up in the cryostat if they have been stored at -70 or -80 degrees.  If they are 
too cold they are brittle too.  

I have not tried the tape method.  We just place the sections directly on the 
slide.  

Paula

Sent from my iPhone

> On Apr 15, 2021, at 11:24 AM, Heather Deziel  wrote:
> 
> 
> Hi Paula,
> 
> I am cutting at -24, but have tried going as warm as -18.  I am currently 
> learning with 30uM sections with the ultimate goal of moving towards 5 or 
> 10uM.  Our lab standard has been collecting into millonigs solution and doing 
> free floating IHC.  We generally have no issue with this technique, but do 
> lose some of the peripheral damaged tissue near the infarct in our stroke 
> brains.  We're trying to work up painting the samples directly onto slides 
> and skipping free floating staining to get a better end product.
> 
> My current samples are very old, they were collected into 4%PFA in 2017 and 
> cryoprotected by freezing in OCT cryomatrix in a little dish floating on LN2 
> in 2019. They've been stored at -80 since then.  Usually we process the 
> brains within a few weeks of collecting them so this particular tissue isn't 
> our ideal situation. We're using old tissue to practice technique, so the 
> current samples aren't going to be used for any actual analysis.  When we 
> heard about the tape method of collecting we were very curious as the the 
> opinion other labs have about it.  Have you tried it?
> 
> Thanks for the answer!
> 
> Heather
> 
> Heather Deziel, MSc.
> Laboratory Technician, CNS|CRO
> 550 University Ave, Charlottetown, PE C1A 4P3
> 
>   (a subsidiary of Neurodyn Life Sciences Inc.)
>  
> 
> 
>> On 2021-04-15 10:29, Patpxs wrote:
>> 
>> Good Morning Heather,
>> 
>> I have some  questions about how you cut frozen brains.   
>> 
>> What temperature are you cutting at?
>> How thick are your sections?
>> How are your samples frozen?  Flash freezing, slow freezing, iso-pentane in 
>> LN2? 
>> 
>> Your answers may provide clues to help you get better cryosections. 
>> 
>> Paula
>> 
>> Sent from my iPhone
>> 
>>> On Apr 15, 2021, at 5:39 AM, Heather Deziel via Histonet 
>>>  wrote:
>>> 
>>> Hello Histonet, 
>>> 
>>> I'm looking into working up a tape transfer method of collecting
>>> cryosections of brain while preserving infarct to be used in IHC.  I
>>> find that when I try and section heavily damaged regions of the brain
>>> the tissue tears and and I lose it.  Has anyone got any recommendations
>>> about the the Section-lab transfer tape (Kawamoto method), using the
>>> circuit plating tape recommended here
>>> (https://www.future-science.com/doi/full/10.2144/btn-2018-0021) or the
>>> cryojane system from Leica?
>>> 
>>> Thank you, 
>>> 
>>> Heather
>>> 
>>> Heather Deziel, MSc.
>>> 
>>> Laboratory Technician, CNS|CRO 550 University Ave, Charlottetown, PE C1A
>>> 4P3 
>>> 
>>>  (a subsidiary of Neurodyn Life Sciences Inc.)
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Re: [Histonet] Filed slides sticking together

[Patpxs via Histonet]

We used to use a drying oven set at 37 degrees C.  We stopped using it because 
some people thought it smelled, plus our slide volume increased to the point 
where all the flats couldn’t fit.  

Now we dry them for two days, minimum, before filing.  I prefer to let them dry 
for at least 4 days.  

Paula

Sent from my iPhone

> On Apr 16, 2021, at 5:26 PM, John Garratt via Histonet 
>  wrote:
> 
> Use tape if you want to have a lean mean operation. Instant filing, no 
> sticking, no ovens.
> 
> John
> 
>> On Fri, Apr 16, 2021 at 2:55 PM, Jay Lundgren via Histonet 
>>  wrote:
>> 
>> I've seen labs that do this, like 30C for 24 hours, but if you use xylene,
>> make sure to put the oven under a hood.
>> 
>> It's a lot easier just to leave them out, in a well ventilated place, for
>> 48hours. Buy more slide folders/trays.
>> 
>> I worked one place where the slides were held for 24 hours before being
>> given to the paths, because they were concerned about being exposed to
>> xylene fumes. But that lab/hospital system had no competition within 100
>> miles and didn't care about turn around times.
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Re: [Histonet] Tape Transfer Methods For Cryosectioned Brains

[Patpxs via Histonet]

Good Morning Heather,

I have some  questions about how you cut frozen brains.   

What temperature are you cutting at?
How thick are your sections?
How are your samples frozen?  Flash freezing, slow freezing, iso-pentane in 
LN2? 

Your answers may provide clues to help you get better cryosections. 

Paula

Sent from my iPhone

> On Apr 15, 2021, at 5:39 AM, Heather Deziel via Histonet 
>  wrote:
> 
> Hello Histonet, 
> 
> I'm looking into working up a tape transfer method of collecting
> cryosections of brain while preserving infarct to be used in IHC.  I
> find that when I try and section heavily damaged regions of the brain
> the tissue tears and and I lose it.  Has anyone got any recommendations
> about the the Section-lab transfer tape (Kawamoto method), using the
> circuit plating tape recommended here
> (https://www.future-science.com/doi/full/10.2144/btn-2018-0021) or the
> cryojane system from Leica? 
> 
> Thank you, 
> 
> Heather
> 
> Heather Deziel, MSc.
> 
> Laboratory Technician, CNS|CRO 550 University Ave, Charlottetown, PE C1A
> 4P3 
> 
>  (a subsidiary of Neurodyn Life Sciences Inc.)
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Re: [Histonet] Antibody diluent

[Patpxs via Histonet]

I’m not completely sure but I think you can do either a mini validation or a 
comparison validation to show there’s no difference between the results.   

I think CAP has a requirement for what to do when reagents change.  

Paula

Sent from my iPhone

> On Mar 25, 2021, at 6:19 AM, Jeanine Ronkowski via Histonet 
>  wrote:
> 
> Scott:I’m pretty certain that you wouldn’t need to completely re-validate 
> after changing diluents, but you may have to change the dilution factor 
> slightly.  It’s my understanding that a change like this only requires 
> staining a few known positive samples, verifying that the results you obtain 
> are very similar if not identical to previous results.  While taking someone 
> else’s suggestion to download that paper on diluents from Biocare’s site, I 
> came across a paper on the subject of validation/verification that I found 
> interesting.Jeanine
> 
> 
> Sent from Yahoo Mail for iPhone
> 
> 
> On Thursday, March 25, 2021, 8:30 AM, Lindrud, Scott 
>  wrote:
> 
> What is everyone's take on what needs to be done from a 
> verification/validation standpoint if you switch diluents for your 
> concentrated antibodies?  The CAP summary of recommendations on the 
> validation IHC antibodies doesn't specifically mention the changing of 
> diluent as one of the variables.
> 
> If I change the diluent on my ER and Her2 concentrates, do I need to do a 
> complete re-validation or just a quick verification using a few positive and 
> negative cases?  I'm not really liking the idea of a complete re-validation!
> 
> Thanks for any info!
> 
> Scott
> 
> 
> Scott A. Lindrud, MLSCM(ASCP)CTCM  | Histopathology Technical 
> Specalist/Cytotechnologist
> 
> P: 320-231-4520
> F: 320-231-4503
> scott.lind...@carrishealth.com
> 
> Rice Memorial Hospital
> 301 Becker Ave SW
> Willmar, MN 56201
> 
> 
> 
> 
> -Original Message-
> From: Jeanine Ronkowski 
> Sent: Tuesday, March 23, 2021 2:21 PM
> To: Paula Keene Pierce ; Maria Cruz 
> ; Morken, Timothy 
> Cc: Histonet 
> Subject: Re: [Histonet] Antibody diluent
> 
> All:Our lab uses a ton of concentrated antibodies and we’ve found that 
> there’s really no such thing as a universal diluent like the ones offered by 
> Leica and Agilent.  We get ours from Biocare, they offer five different 
> diluents with different pHs and chemical makeups and we use three of these. I 
> checked with my Biocare rep and iit seems that the one that’s most similar to 
> Leicas is called Monet Blue.  I suggest you contact your Bocare rep and see 
> if they can send you a sample of one or more them. Good  luck,Jeanine
> 
> 
> Sent from Yahoo Mail for iPhone
> 
> 
> On Tuesday, March 23, 2021, 2:43 PM, Paula Keene Pierce via Histonet 
>  wrote:
> 
> I just purchased antibody diluent from Electron Microscopy Sciences.
> Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
> Blue Lake DriveNorman, OK 73069PH 
> 405-759-3953http://www.excaliburpathology.com
> 
> A sharp knife is nothing without a sharp eye. - Klingon Proverb
> 
> On Tuesday, March 23, 2021, 01:36:12 PM CDT, Morken, Timothy via Histonet 
>  wrote:
> 
> Maria, we went back to using Dako Diluent, which we had just switched away 
> from to save money!
> 
> Tim Morken
> Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
> Pathology UC San Francisco Medical Center
> 
> -Original Message-
> From: Maria Cruz via Histonet 
> Sent: Tuesday, March 23, 2021 10:55 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Antibody diluent
> 
> Like other folks in the histo community, I’ve recently experienced the 
> inability to buy diluent from Leica.  Now wondering what others are doing to 
> handle this problem.  TIA!
> Maria
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Re: [Histonet] Warm formalin

[Patpxs via Histonet]

Hi Erin,

Often heat is applied to formalin to speed up fixation.   That said there is 
probably a temperature point where it goes from fixing tissue to cooking it.  

Paula

Sent from my iPhone

> On Jul 21, 2020, at 6:14 PM, Martin, Erin via Histonet 
>  wrote:
> 
> Hello everyone!
> 
> We have a referring clinician that is concerned about leaving his specimens 
> in an outdoor lockbox in the summer because the formalin will get hot.  I 
> don't think that having some specimens in formalin in hot weather would cause 
> any problems but I can't find any references one way or another.  Does anyone 
> have any policies regarding this?
> 
> Thanks so much!
> 
> 
> Erin Martin, Histology Supervisor
> 
> UCSF Dermatopathology & Oral Pathology Service
> 
> Phone: 415-3537248 | Fax: 415-353-7543
> 
> 
> CONFIDENTIALITY NOTICE: This email message, including any attachments, is for 
> the sole use of the intended recipient(s) and may contain confidential, 
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> message in error, please contact the sender by reply email and destroy all 
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Re: [Histonet] Tissue processor errors, failures and what to do

[Patpxs via Histonet]

Hi Garrey,

The answer is “it depends”.   What you do when a processor fails depends on the 
failure point.  If the tissue is still in dehydrant it gets treated differently 
than if it fails in the intermediate solvent.  

Paula

Sent from my iPhone

> On Jul 4, 2020, at 10:08 AM, Garrey Faller via Histonet 
>  wrote:
> 
> Happy 4th to all.
> Does anyone have a procedure on what to do when a tissue processor fails or 
> alarms.  I want to learn more about the science behind tissue processing so I 
> know what to do when the machine fails. This happened to a friend recently 
> and I want to prevent my tissues/biopsies from being ruined.
> Thanks.
> Garrey
> 
> Sent from my iPhone
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Re: [Histonet] Tissue processing

[Patpxs via Histonet]

I favor the VIP too.  My lab has 4 VIPs and 1 Leica Peloris.   The VIPs can 
process more cassettes on a run and cost a lot less than the Peloris.  

Paula

Sent from my iPhone

> On Mar 6, 2020, at 12:34 AM, warda hassan via Histonet 
>  wrote:
> 
> Hello to all histonets
> 
> Advanced thanking to whoever will give me a feedback on their selection for
> tissue processing
> We are to purchase a new tissue processing
> List are
> Histo pro 200 & 300( histoline)
> Milstone
> Diapth
> Shandon - Thermo Revos
> Sakura VIP TEK6
> 
> Any feedback on your hand-on experiences with these system will be
> highly  appreciated
> 
> Manythanks
> 
> Rose
> Histo-Incharge
> Royal Hospital
> Oman
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Re: [Histonet] Changing to reagent alcohol

[Patpxs via Histonet]

Hi Carole,

Sorry but you need to validate the change completely.  Reagent alcohols are 
blends that sometimes don’t even contain ethanol.   You need to demonstrate 
that it doesn’t affect routine H, special stains, and IHC.   Plus you have 
to test the major tissue types you routinely work on.  

Others may disagree with me, but if you get an inspector who focuses on the 
minutia, you can get cited. 

Paula

Sent from my iPhone

> On Feb 6, 2020, at 12:13 PM, Johnson, Carole via Histonet 
>  wrote:
> 
> We have been using ethanol in our lab and would like to change to using 
> reagent alcohol for the obvious reasons of cost and regulatory headaches. I 
> have been tasked with creating a validation/verification process.  Does 
> anyone have any suggestions?  Or better yet, information proving that we can 
> make the change without going through an extensive project?  Thanks!
> 
> Carole L Johnson, HT(ASCP)cm, QIHC(ASCP)cm
> Histotechnologist
> UCH/Memorial Central Hospital
> Histology Laboratory
> 1400 Boulder Street
> Colorado Springs, CO
> O 719.365.5204
> carole.john...@uchealth.org
> 
> CONFIDENTIALITY NOTICE: This message and any attachments may contain 
> privileged and confidential peer review, risk management, and/or quality 
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Re: [Histonet] New York Qualifications for Histology Supervisor

[Patpxs via Histonet]

This is an issue that continues to pop up.   It all starts with the CLIA 
regulations that do not recognize histology professionals as equal to other 
laboratory professionals.  Until the feds change CLIA we will  keep having this 
problem.  

I do believe that NSH is petitioning on our behalf to change the CLIA 
regulations.   

In California the state does not recognize Histotechs, certified or not, so we 
can’t get licensed and we can’t become lab managers as a result.  All thanks to 
CLIA.  

That’s my rant for the day.  

Paula

Sent from my iPhone

> On Feb 21, 2019, at 8:58 AM, Colleen Forster via Histonet 
>  wrote:
> 
> Wow...this is just crazy~ The regulations leave histology in a real tough
> spot...
> 
> Feeling for all of you and yes, for the future histology techs coming up~
> 
> Colleen Forster
> U of MN
> 
> On Thu, Feb 21, 2019 at 10:52 AM Perl , Alison via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> 
>> Hi Melissa
>> This is true, and a source of my neverending frustration with NYS Office
>> of Professions. Stephanie Shulman (stephanie.shul...@health.ny.gov) is a
>> good person at NYS to get clarification, but she will reiterate the same.
>> 
>> NYS has explained to me that "histotechs don't exercise judgment" and thus
>> are not qualified to take on the role of supervisor, even for a
>> histology-only laboratory. The supervisors here in my lab and myself all
>> have Clinical Lab Technologist licenses, from when they were grandfathered
>> back in 2007. I have techs who went to school specifically for Histology,
>> but can never become supervisors. I don't know what anyone will do for the
>> next generation of supervisors - your situation is a real fear.
>> 
>> Anyone in NYS who would like to raise this issue with your
>> representatives, I encourage you to do so!!
>> 
>> Alison Perl, HTL(ASCP)CM
>> Anatomic Pathology Manager
>> CareMount Medical
>> (914) 302-8424
>> ap...@cmmedical.com
>> 
>> 
>> -Original Message-
>> From: Melissa Owens via Histonet [mailto:histonet@lists.utsouthwestern.edu]
>> 
>> Sent: Thursday, February 21, 2019 11:21 AM
>> To: histonet@lists.utsouthwestern.edu
>> Subject: [EXTERNAL] [Histonet] New York Qualifications for Histology
>> Supervisor
>> 
>> Hello,
>> 
>> I have a question about the requirements to be a supervisor in New York.
>> New York dept. of Ed states qualifications for a Medical
>> Technologist/Laboratory Supervisor and a Cytotechnologist/Cytology
>> Supervisor but no where does it mention qualifications for a
>> Histototech/Histology Supervisor. So therefore, I have heard that in New
>> York, a Histology Supervisor must qualify under the Medical
>> Technologist/Laboratory Supervisor qualifications statues. This seems
>> impossible to me as this would have to be an individual who becomes a
>> Medical Technologist and by random transfers into a histology role then
>> becomes a supervisor. Does anyone out there in New York legally qualify
>> your histology supervisors who have a bachelors degree but were not former
>> Medical Technologists? Thank you for any help as I have intensely scoped
>> the New York department of Education website in my frustrations.
>> 
>> By the way, I am asking this because I have a Day Shift 7:30am-4pm
>> Monday-Friday Histology Supervisor position in the New York Metropolitan
>> area with these requirements and I don't know if there is a way around
>> them. Thank you!
>> 
>> Melissa Owens, CHP (ASA)
>> President, Laboratory Staffing
>> Allied Search Partners
>> 
>> 
>> 
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>> part and whether electronically, written and/or orally) and/or taking of
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> 
> 
> -- 
> Colleen Forster HT(ASCP)QIHC
> BLS Histology and IHC Laboratory
> B173 PWB  612-626-1930
> 
> *If submitting histology request please also forward to Lori Holm at
> ho...@umn.edu *
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Re: [Histonet] p2y12 cryo rat problem high background

[Patpxs via Histonet]

It could be the concentration of the antibody is too high.  Have you tried a 
lower dilution?

What species made the antibody?  I know that sounds a bit basic but I know that 
I have used a mouse or rat antibody by mistake.  

Paula

Sent from my iPhone

> On Nov 15, 2018, at 4:44 AM, Kooijman, E.J.M. (Esther) via Histonet 
>  wrote:
> 
> Hello Bobbie,
> 
> But should I elimination endogenous peroxidase activity in brain/spinal cord 
> tissues?
> Brains and spinal cord was harvested after cervical dislocation, then the 
> tissue was snap frozen (isopentane..).
> 
> thanks for your help,
> Esther
> 
> -Oorspronkelijk bericht-
> Van: Boyce, Bobbie [mailto:bobbie.bo...@nemours.org] 
> Verzonden: donderdag 15 november 2018 12:21
> Aan: Kooijman, E.J.M. (Esther)
> Onderwerp: RE: p2y12 cryo rat problem high background
> 
> Hi Ester,
> Try Peroxoblock (Zymed) before your BSA block, but you have to be careful not 
> to leave it on too long or it will eat your tissue. It's been a while since 
> I've had to used it.
> 
> 
> Bobbie Boyce
> Histology Specialist III
> DuPont Experimental Station
> Nemours- Biomedical Research Department
> Histochemistry and Tissue Processing Core
> 200 Powder Mill Road, Bldg.400  Rm.5240
> Wilmington, DE 19803
> 
> (lab) 302-651-6771
> (fax) 302-651-5010
> 
> 
> 
> -Original Message-
> From: Kooijman, E.J.M. (Esther) via Histonet 
>  
> Sent: Thursday, November 15, 2018 5:53 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] p2y12 cryo rat problem high background
> 
> **This is an External Email -  Please DO NOT open attachments or click links 
> from unknown senders or unexpected email. **
> 
> Hello all,
> 
> 
> 
> I am trying to stain cryo brain sections from the rat 7um but having a lot of 
> background. What am I doing wrong. Below the protocol is used and tried to 
> adjust…
> 
> 
> 
> 1-  Fix the tissue with cold acetone (-20 ⁰C) for 10 minutes
> 
> 
> 
> 2-  Let the slide dry for 30 minutes at room temperature. Isolate the 
> sections with DAKO pen
> 
> 
> 
> 3-  Block sections with BSA 2% in PBS for 1 hour at room temperature
> 
> 
> 
> 4-  Add primary antibody in PBS/BSA 1%, overnight at 4⁰C or 1h at room 
> temperature
> 
> 
> 
> 5-  Wash 3x5 minutes with PBS/tween-20 0.05%
> 
> 
> 
> 6-  Add 100 µL Envision solution (goat anti-mouse/rabbit) and incubate 
> for 1hr at RT
> 
> 
> 
> 7-  Wash 3x 5 minutes with PBS/tween-20 0.05%
> 
> 
> 
> 8-  Incubate with DAB (1:50) for 10 min (between 5-10 min; check color 
> development)
> 
> (wear gloves, carcinogenic!)
> 
> 
> 
> 9-  Rinse thoroughly with miliQ water
> 
> 
> 
> 10-   Stain with haematoxylin for 1 min
> 
> 
> 
> 11-   Wash with running tap water for 5 min
> 
> 
> 
> 12-   Start the alcohol/xylene series (70% EtOH -> 80% EtOH -> 96% EtOH -> 
> 100% EtOH -> 100% EtOH/xylene -> xylene -> xylene)
> 
> 
> 
> 13-   Mount slides with entallan
> 
> Kind regards,
> 
> 
> 
> 
> 
> Esther Kooijman  |  Research Technician  |  Department of Radiology and 
> Nuclear medicine
> 
> The Netherlands
> 
> 
> 
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Re: [Histonet] Microscope objectives

[Patpxs via Histonet]

Also, the manufacturers make the threads different so what works on one brand 
won’t work on another.  They even change the threading between models so you 
have to buy new objectives.  

Paula

Sent from my iPhone

> On Jun 19, 2018, at 6:44 AM, Rene J Buesa via Histonet 
>  wrote:
> 
> Not all objectives "are created equally"."In the beginning" (as in Genesis) 
> the objective "was created" to be used on microscopes with essentially two 
> different focal distances, i.e. 160mm and 170mm. By the 1950's the ∞ 
> corrected objective appeared to simplify somewhat the situation BUT every 
> manufacturer tried that all their objectives were "par-focal" meaning that 
> you could switch from one objective to the next and the virtual image 
> remained "in focus".That is the crux of the issue: a virtual image 
> permanently in focus, and that cannot be obtained between objectives from 
> different manufacturers because their sizes (lengths) are different, and 
> their focal points may also differ.So, putting it simply, unless you are 
> willing to refocus with either your "macro" or "micro" knob (focusing wheel) 
> you will need to buy all your objectives from the same manufacturer 
> (brand).René 
> 
>On Tuesday, June 19, 2018 8:58 AM, Charles Riley via Histonet 
>  wrote:
> 
> 
> Are all microscope objectives universal between brands or do you have to
> buy the objectives from the company that manufactured it?
> 
> -- 
> 
> Charles Riley BS  HT, HTL(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
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Re: [Histonet] RELIA Histology Careers Bulletin - Live where you vacation 8-1-2017

[Patpxs via Histonet]

My comment was meant in jest.   I live in San Diego, so Modesto is not a city I 
would vacation.  Modesto probably has many nice features.   

Paula

Sent from my iPhone

> On Aug 2, 2017, at 6:32 AM, Pam Barker via Histonet 
>  wrote:
> 
> Hi Histonetters!
> I just want to take a minute to address the opinion expressed about one of
> the employers on my most recent email.
> I apologize that there was editorializing on a particular lab and I just
> want to point out after over 30 years in recruiting in all types of
> environments in all fairness: 
> The quality of the workplace is in the eye of the beholder.  
> In other words not every place is right for everybody.  
> Not every place is right for everyone but I promise that I will work my
> hardest to find you the right position in the right place.
> 
> Thanks again - Pam
> 
> 
> Thanks-Pam
> 
> Right Place, Right Time, Right Move with RELIA!
> 
> Thank You!
> Pam M. Barker
> 
> Pam Barker
> President/Senior Recruiting Specialist-Histology
> RELIA Solutions
> Specialists in Allied Healthcare Recruiting
> 5703 Red Bug Lake Road #330
> Winter Springs, FL 32708-4969
> Phone: (407)657-2027
> Cell: (407)353-5070
> FAX: (407)678-2788
> E-mail: rel...@earthlink.net 
> https://www.facebook.com/RELIASolutionsforhistologyprofessionals
> www.facebook.com  /PamBarkerRELIA
> www.linkedin.com/in/reliasolutions
> www.twitter.com/pamatrelia 
> 
> 
> 
> 
> 
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Re: [Histonet] Water for H Stainers?

[Patpxs via Histonet]

And the winner for the water of choice for H strainers is..

What ever you have available.  Tap, tap with a filter, deionized.  

It all depends on the water quality in your area.  

Thanks to all who replied. 

Paula

Sent from my iPhone

> On Sep 27, 2016, at 9:54 AM, Elizabeth Chlipala  wrote:
> 
> Paula
> 
> We have ours hooked up to tap water.
> 
> Liz
> 
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Premier Laboratory, LLC
> PO Box 18592
> Boulder, CO 80308
> (303) 682-3949 office
> (303) 682-9060 fax
> (303) 881-0763 cell
> l...@premierlab.com
> www.premierlab.com
> 
> Ship to Address:
> 
> Premier Laboratory, LLC
> 1567 Skyway Drive, Unit E
> Longmont, CO 80504
> 
> -Original Message-
> From: P Sicurello via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
> Sent: Tuesday, September 27, 2016 10:28 AM
> To: HistoNet
> Subject: [Histonet] Water for H Stainers?
> 
> Good Morning Everyone,
> 
> 
> 
> What type of water do you use with your automated H stainers?  House or 
> deionized/distilled?
> 
> 
> 
> We cannot buy one of the waterless, fancy schmancy H stainers at this time. 
>  L Thank you in advance.
> 
> Sincerely,
> 
> 
> 
> Paula
> 
> 
> 
> Paula Sicurello, HTL (ASCP)CM
> 
> Histotechnology Specialist
> 
> UC San Diego Health
> 
> 200 Arbor Drive
> 
> San Diego, CA 92103
> 
> (P): 619-543-2872
> 
> 
> 
> *Confidentiality Notice*: The information transmitted in this e-mail is 
> intended only for the person or entity to which it is addressed and may 
> contain confidential and/or privileged material.  Any review, retransmission, 
> dissemination or other use of or taking of any action in reliance upon this 
> information by persons or entities other than the intended recipient is 
> prohibited.  If you received this e-mail in error, please contact the sender 
> and delete the material from any computer.
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Re: [Histonet] Freeze Spray Not Sold as Case Happy Friday

[Patpxs via Histonet]

I heard everything on the intervention is true, is that true?

Paula ;-)

Sent from my iPhone

 On Aug 21, 2015, at 9:16 AM, Jay Lundgren via Histonet 
 histonet@lists.utsouthwestern.edu wrote:
 
 There's this cool thing called the Interwebz now?
 
 http://www.amazon.com/Multi-Purpose-Freeze-Spray-1-Ea/dp/B0017UIB30#Ask
 
  Sincerely,
 
 Jay A. Lundgren, M.S., HTL (ASCP)
 
 On Wed, Aug 19, 2015 at 8:53 PM, ian bernard via Histonet 
 histonet@lists.utsouthwestern.edu wrote:
 
 Fellow Histonetters: I'm looking for a source to purchase Freeze-Spray used
 during the frozen procedure.  Our current resource sells Freeze Spray in a
 case of 6 cans. I would like to purchase as an individual can owing to our
 workload rather than a case.
 
 
 
 Any references?
 
 
 
 Ian Bernard
 
 
 
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Re: [Histonet] Slides stuck together

[Patpxs via Histonet]

 Hi Tonya,

Soak the brick-o-slides in xylene until they unstick from each other. 

This can take over a week, you must be patient. The slides will break if you 
try to force them apart. 

Paula

Sent from my iPhone

 On Aug 10, 2015, at 6:51 AM, Abbott, Tanya via Histonet 
 histonet@lists.utsouthwestern.edu wrote:
 
 We found a batch of GYN slides that were filed, and have now bonded 
 together. We use the Sakura coverslipper, so we are told this is not supposed 
 to happen.
 I wonder if it is from residual glue from the coverslipping process? Anyone 
 have any suggestions as to how to get these slides unbonded?
 Thanks in advance!
 Tanya
 
 Tanya G. Abbott
 Manager Technologist
 Histology/Cytology
 Penn State Health St. Joseph
 (phone) 610-378-2635
 
 This email and attachments contain information that may be confidential or 
 privileged. If you are not the intended recipient, notify the sender at once 
 and delete this message completely from your information system. Further use, 
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Re: [Histonet] Ventana Retic Stain Quality

[Patpxs via Histonet]

I was told by the technical specialist to make the wash solution fresh before 
use. There has been a contamination caused by growth of organisms in the wash 
if it sits around too long between uses. 

Paula :-)

Sent from my iPhone

 On Jul 16, 2015, at 10:14 AM, Blake Taylor via Histonet 
 histonet@lists.utsouthwestern.edu wrote:
 
 Are you using the Benchmark Special Stainer?  If so it is due to a 
 containment in the wash solution.  Ventana knows they have a problem and are 
 currently beta testing a new wash solution at other sites that should be 
 rolled out soon. Of course they said soon 6-9 months ago.  Anytime our Retic 
 starts to have an issue we have to decon the machine.  We have had our 
 technical specialist in here numerous times and they are fully aware.
 
 
 
 -Original Message-
 From: histonet-requ...@lists.utsouthwestern.edu 
 [mailto:histonet-requ...@lists.utsouthwestern.edu] 
 Sent: Thursday, July 16, 2015 1:00 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: Histonet Digest, Vol 140, Issue 18
 
 Send Histonet mailing list submissions to
histonet@lists.utsouthwestern.edu
 
 To subscribe or unsubscribe via the World Wide Web, visit
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 or, via email, send a message with subject or body 'help' to
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 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. HE stainers (Fawn Bomar)
   2. Medical necessity question (Trish Hicks)
   3. RELIA HOT HOT HOT Histology Job ALERT A RELIA EXCLUSIVE
  Histology Lab Manager - Dallas, TX (Pam Barker)
   4. test (Amy Self)
   5. Fw: Re:  : Ventana Retic Stain Quality (Kiranjit Grewal)
   6. Re: Ventana LCS (Liquid Cover Slip)/ non-specificstaining
  (Burnett, Brandy)
 
 
 --
 
 Message: 1
 Date: Wed, 15 Jul 2015 17:31:30 +
 From: Fawn Bomar fawn.bo...@halifaxregional.com
 To: histonet@lists.utsouthwestern.edu
histonet@lists.utsouthwestern.edu
 Subject: [Histonet] HE stainers
 Message-ID:
35b63a2e2fc1c8429d3acf1cdda5ffca05e...@exch-2k10.hrhs.com
 Content-Type: text/plain; charset=iso-8859-1
 
 Hi everyone,
 
 
 
 I know this question has been asked before but I am having difficulty finding 
 it.  We are in the process of finding a new HE stainer and was wondering 
 what everyone would recommend or not recommend.  We are a small facility- 
 stain less than 100 HE slides a day.  We are also looking for one that is 
 compatible with a coverslipper.  Right now my manager is looking at the 
 Sakura Prisma but we would have to do some major reconstruction to fit it in 
 our lab.  We are also looking at the Gemini with the Clearvue coverslipper.  
 Can anyone give me their opinions on what they think?
 
 
 
 Thank you
 
 Fawn
 -
 This electronic message may contain information that is confidential or 
 legally privileged.  It is intended only for the use of the individual(s) and 
 entity named as recipients in the message. 
 
 If you are not an intended recipient of this message, please notify the 
 sender immediately and delete the material from any computer. Do not deliver, 
 distribute, or copy this message, and do not disclose its contents or take 
 any action in reliance on the information it contains. 
 
 Thank you
 
 
 --
 
 Message: 2
 Date: Wed, 15 Jul 2015 17:42:12 +
 From: Trish Hicks thi...@gulfcoastdermpath.com
 To: histonet@lists.utsouthwestern.edu
histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Medical necessity question
 Message-ID:
96d42d0e9fb88a46a1b3e3e82de4ca9a5627a...@gcd-exch.tampa.gcd.local
 Content-Type: text/plain; charset=us-ascii
 
 We are under the impression that we will be required to start putting a 
 reason that a stain or IHC stain was performed in the microscopic 
 description, or somewhere, for CMS to consider paying for an additional 
 stain.  It is a requirement that is upcoming and we want to be proactive.  
 How is everyone else handling this or is what already appears in the 
 microscopic description or diagnosis sufficient?
 
 Thank You,
 
 Trish Hicks
 BS, HTL (ASCP)
 Laboratory Supervisor
 Gulf Coast Dermatopathology Laboratory, Inc.
 Ph: 813-882-4206
 Fax: 813-886-0589
 Toll Free: 877-654-7721
 thi...@gulfcoastdermpath.com
 Confidentiality: The information transmitted is intended only for the person 
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