Re: [Histonet] (no subject)

[Tony Reilly via Histonet]

Hello Rhonda
I have extensive experience with the Brown Hopps stain.

I would advise against only staining for gram negative as both the gram 
positive stain and the gram negative stain will stain both gram positive and 
gram negative organisms.  The differentiation is only achieved by the 
destaining steps.

If you only used the gram negative steps you would still be staining both gram 
positive and gram negative organisms negatively.

If you you want more information let me know .

Regards
Tony Reilly
Histologist
Australia



Sent from my iPhone

> On 4 Jul 2023, at 5:30 am, Mac Donald, Jennifer via Histonet 
>  wrote:
> We have good success with the Twort.
> 
> -Original Message-
> From: Rhonda McCormick via Histonet 
> Sent: Monday, July 3, 2023 11:22 AM
> To: Histonet 
> Subject: [Histonet] (no subject)
> 
>  EXTERNAL SENDER - Exercise caution with requests, links, and attachments.
> 
> Hello, I'm wondering if anyone does the Brown-Hopps Gram stain for gram 
> negative bacteria. If so, would you mind to send your protocol please?
> Any recommendations for a gram negative (only) stain - specifically E. coli? 
> One of our residents is asking.
> Thank you!
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Re: [Histonet] Alcian blue van gieson double stain

[Tony Reilly via Histonet]

Hi Charles

I have done this stain many times as an abbreviated version of Movats 
Pentachrome.
You are correct in thinking the van gieson will affect the AB stain due to the 
acid.

As in the Movat stain do AB first then place slides in alcoholic ammonia for 1 
hour to convert AB to insoluble Monastral blue then finish with the VVG.

Regards
Tony Reilly

Sent from my iPhone

> On 10 Mar 2023, at 5:49 am, Akemi Allison via Histonet 
>  wrote:
> 
> Just curious, why not do a Movat’s Pentachome? It gives you everything you 
> are looking for and more.  
> Best regards,
> Akemi Allison-Tacha, BS, Ht, HTL 
> akemiat3...@gmail.com
> 
> Sent from my iPhone
> 
>> On Mar 9, 2023, at 11:19 AM, Paula Keene Pierce via Histonet 
>>  wrote:
>> 
>> Hi,
>> if they are wanting to stain elastic, Alcian blue, and Van Gieson, I 
>> recommend doing the Verhoff elastic stain first, then the Alcian blue, and 
>> then the Van Gieson.
>> 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 Thursday, March 9, 2023 at 01:05:14 PM CST, Charles Riley via Histonet 
>>  wrote:  
>> 
>> I am working with a researcher and they saw an article where someone
>> performed an Alcian Blue  and elastic van Gieson double stain.  If anyone
>> has done this before can you provide your protocol.  I am trying to figure
>> out the best way to perform this process to get the best staining results.
>> Here are my questions/thoughts
>> 
>> 1. Is it better to run the Alcian blue as usual and then perform the van
>> gieson stain?
>> 2. Will the differentiating solution in the van gieson stain affect the
>> alcian blue staining in any way?
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Re: [Histonet] destain for IHC

[Tony Reilly via Histonet]

Hi

If you are using the peroxidase method the hydrogen peroxide step will remove 
the eosin and some of the haematoxylin.

No need to do a decolorisation step before staining.

As Gudren said the only problem may be not having coated slides.

Good luck.
Sadly retired histologist
Tony

Sent from my iPhone

> On 10 Feb 2023, at 6:07 am, Katleba, Maria via Histonet 
>  wrote:
> 
> Oh no!!! We cut all slides on + 
> 
> BUT you can do this. It’s what we do when we have H on plain slides.
> 
> Whether  you have a dry unstain from another lab that sent u a slide, or your 
> H that you just took the coverslip off…
> 
> Place it in. Coplin jar with 1-3 DROPS of formalin, cover, place In 60ft 
> degree oven for 60 min. You want the bottom of the jar to be moist not 
> floating in it 
> 
> After 60 min remove and open UNDER A HOOD (it’s a vaporized carcinogen)
> Air out 10-15 min… place on stainer 
> 
> The drops vaporize and the aldehyde goes through tissue and make it adhere to 
> the slide permanently.
> 
> I do this when I cut nails or anything that won’t stay on a plus slide.
> 
> Maria
> 
> Sent from my iPhone
> 
>> On Feb 8, 2023, at 11:21 PM, Gudrun Lang via Histonet 
>>  wrote:
>> 
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Re: [Histonet] Processing Protocols

[Tony Reilly via Histonet]

Hi Jessica

My advice would be to process your small biopsies during the day allowing 
depending on the times you use at least 2 runs per day.  One in the am and one 
in the pm.  Then run your large blocks over night as per usual.  Any late 
arriving small biopsies can be run on the am run the following day .  This way 
everything is processed within 24 hours of arrival maintaining your TATs.

Regards

Tony
Retired Histotragic

Sent from my iPhone

> On 4 Aug 2021, at 4:46 am, Terri Braud via Histonet 
>  wrote:
> 
> We have encountered a similar problem with one of our two processors is 
> down.  Shortening the longer schedule produced underprocessed fatty tissues 
> and over processed biopsies, so we run the biopsies on the short schedule 
> overnight, run a quick clean cycle, then the larger tissues on the 10hr 
> schedule on the same instrument, pull the them off and leave them in the 
> embedding unit to embed in the afternoon, or next morning.  It does put the 
> larger tissue a day behind, but does not sacrifice the quality of the 
> processing.  It you shorten your large tissue processing run so that you can 
> run small biopsies with the bigger stuff, you will have to revalidate your 
> IHC, so maybe this might be a better alternative.  Good luck, Terri
> 
> Terri L. Braud, HT(ASCP)
> HNL Laboratories for 
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> Ph: 215-938-3689
> Fax: 215-938-3874
>   Honesty
> AccouNtability
> AgiLity
> CoLlaboration
>   CoMpassion
> 
> Message: 1
> Date: Tue, 3 Aug 2021 10:34:11 +
> From: "Piche, Jessica" 
> Subject: [Histonet] Processing protocols
> 
> Good Morning Everyone,
> Our pathologist wants us to shorten our large tissue processing run so that 
> we can run small biopsies with the bigger stuff when one of our tissue 
> processors are down. We have 2 Excelsiors. Would anyone like to share any 
> protocols they might have for this purpose?
> Thank you!
> Jessica Piche, HT(ASCP}
> Waterbury Hospital Histology Lab
> 
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Re: [Histonet] Cell block preparations

[Tony Reilly via Histonet]

Some predictive text issues like lyse and nucleated. 

Sent from my iPhone

> On 17 Jan 2020, at 8:24 am, Tony Henwood (SCHN) via Histonet 
>  wrote:
> 
> Hi Jennifer,
> 
> I have had excellent success with lysing the red blood cells (using  Isotonic 
> Ammonium Chloride) prior to cell block preparation with 
> thromboplastin-plasma. 
> The lysing solution contains EDTA so you will need to add a few drops of 1% 
> calcium chloride. Method as follows:
> 
> Lysis solution
> Ammonium Chloride4.5g
> Potassium carbonate  0.5g
> EDTA 0.0186g
> Distilled water  500mls
> 
> Method:
> 1.Centrifuge bloody fluid.
> 2.Remove supernatant and add equal volume of lysis solution.
> 3.Resuspend and incubate for 5 minutes at 4oC.
> 4.Centrifuge, if blood still remains, then repeat from step 2.
> 5. Rinse in Hanks or RPMI, centrifuge.
> 6.Mix pellet in a few drops of plasma.
> 7. Add thromboplastin and a few drops  of 1% Calcium Chloride, mix gently 
> and allow clot to form.
> 8. Add 10% buffered formalin and fix and process as usual.
> 
> Reference:  
> Kuenen-Boumiester etal (1996) Acta Cytolog 40:475-479
> 
> If you donot use the plasma clot method for cell block preparation, then use 
> your preferred method after step 5.
> The lysis solution can also be purchased commercially from several companies 
> (eg Biolegend). It is commonly used for sample preparation for flow 
> cytometry. Check the SDS to make sure it does not contain formaldehyde.
> 
> 
> -Original Message-
> From: Mac Donald, Jennifer via Histonet 
> [mailto:histonet@lists.utsouthwestern.edu] 
> Sent: Friday, 17 January 2020 4:53 AM
> To: Charles Riley ; Histo List 
> 
> Subject: Re: [Histonet] Cell block preparations
> 
> Acetic acid would work.
> 
> Get Outlook for iOS 
> From: Charles Riley via Histonet 
> Sent: Thursday, January 16, 2020 8:55:21 AM
> To: Histo List 
> Subject: [Histonet] Cell block preparations
> 
>  EXTERNAL SENDER- Exercise caution with requests, links, and attachments.
> 
> What is the best way to remove excess blood from FNA sample collections 
> before spinning them down into cell blocks?
> 
> --
> 
> Charles Riley BS  HT, HTL(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
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> information. If you are not the intended recipient, please delete it and 
> notify the sender.
> 
> Views expressed in this message are those of the individual sender, and are 
> not necessarily the views of NSW Health or any of its entities.
> 
> 
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Re: [Histonet] Cell block preparations

[Tony Reilly via Histonet]

Hi Charles

A method my EM scientist used many years ago was quite simplistic and he 
thought it worked well.
You simply centrifuge the specimen,  add a set volume of deionised water, mix 
to lose the red cells, then add an equal volume of double strength normal 
saline to create a suspension in normal saline.  
The centrifuge where the heavier uncleared cells go to the bottom of the tube.  
I have to say the I have never tried this personally but if the morphology was 
retained for EM it must be fine for LM.
Having said that I have great respect for Tony Henwood and his method looks 
much more scientific.

Regards
Tony

Sent from my iPhone

> On 17 Jan 2020, at 8:24 am, Tony Henwood (SCHN) via Histonet 
>  wrote:
> 
> Hi Jennifer,
> 
> I have had excellent success with lysing the red blood cells (using  Isotonic 
> Ammonium Chloride) prior to cell block preparation with 
> thromboplastin-plasma. 
> The lysing solution contains EDTA so you will need to add a few drops of 1% 
> calcium chloride. Method as follows:
> 
> Lysis solution
> Ammonium Chloride4.5g
> Potassium carbonate  0.5g
> EDTA 0.0186g
> Distilled water  500mls
> 
> Method:
> 1.Centrifuge bloody fluid.
> 2.Remove supernatant and add equal volume of lysis solution.
> 3.Resuspend and incubate for 5 minutes at 4oC.
> 4.Centrifuge, if blood still remains, then repeat from step 2.
> 5. Rinse in Hanks or RPMI, centrifuge.
> 6.Mix pellet in a few drops of plasma.
> 7. Add thromboplastin and a few drops  of 1% Calcium Chloride, mix gently 
> and allow clot to form.
> 8. Add 10% buffered formalin and fix and process as usual.
> 
> Reference:  
> Kuenen-Boumiester etal (1996) Acta Cytolog 40:475-479
> 
> If you donot use the plasma clot method for cell block preparation, then use 
> your preferred method after step 5.
> The lysis solution can also be purchased commercially from several companies 
> (eg Biolegend). It is commonly used for sample preparation for flow 
> cytometry. Check the SDS to make sure it does not contain formaldehyde.
> 
> 
> -Original Message-
> From: Mac Donald, Jennifer via Histonet 
> [mailto:histonet@lists.utsouthwestern.edu] 
> Sent: Friday, 17 January 2020 4:53 AM
> To: Charles Riley ; Histo List 
> 
> Subject: Re: [Histonet] Cell block preparations
> 
> Acetic acid would work.
> 
> Get Outlook for iOS 
> From: Charles Riley via Histonet 
> Sent: Thursday, January 16, 2020 8:55:21 AM
> To: Histo List 
> Subject: [Histonet] Cell block preparations
> 
>  EXTERNAL SENDER- Exercise caution with requests, links, and attachments.
> 
> What is the best way to remove excess blood from FNA sample collections 
> before spinning them down into cell blocks?
> 
> --
> 
> Charles Riley BS  HT, HTL(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
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> information. If you are not the intended recipient, please delete it and 
> notify the sender.
> 
> Views expressed in this message are those of the individual sender, and are 
> not necessarily the views of NSW Health or any of its entities.
> 
> 
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