[Histonet] TUNEL on Leica Bond

Fri Oct 19 14:24:01 CDT 2018

Hello Histo-friends,
Is anyone doing a TUNEL stain on any of the Leica Bond instruments?  I will be attempting to set this up and thought I would reach out first.
Thank you,
Cathy

Cathy M. Mathis 
Laboratory Technician IV

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Subject: Histonet Digest, Vol 179, Issue 16

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Today's Topics:

   1. toluene substitute (Michelle Jamison)
   2. RUO antibody (Erin McCarthy)
   3. Re: Restaining old Heamatology smears (Eddie Martin)
   4. Re: BMP-2 protocol for bone (Eddie Martin)

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Message: 1
Date: Thu, 18 Oct 2018 14:48:50 -0600
From: Michelle Jamison <m.jamison at elitechgroup.com>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] toluene substitute
Message-ID: <a388a0e7-91fc-7555-36ac-3d8539d549aa at elitechgroup.com>
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Hi All, I am from industry. We are interested in developing an 
instrument that stains then prepares microscope slides to be 
coverslipped. If you do coverslip, what substitute do you use instead of 
toluene to go to mounting media? Thank you! Michelle
-- 

All the best to you,

Michelle Jamison

/Bio Research Associate /

/ELITechGroup Inc. / ???370 W. 1700 S.?? ???Logan Utah ??? USA

m.jamison at elitechgroup.com <mailto:m.jamison at elitechgroup.com> ? 
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Message: 2
Date: Thu, 18 Oct 2018 16:30:14 -0500
From: Erin McCarthy <erin.mccarthy at tempus.com>
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] RUO antibody
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	<CAOjxy-HdKP0nZxUBX3O8iDijMeho==ro-BP6s6pkQFDmy0o2LA at mail.gmail.com>
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Hi All,

I am looking to see if anyone has had any experience with the antibody AXL.
We are trying to work it up on our Roche Discovery Ultra and it does not
seem like it stains both nuclear and cytoplasic cell features. We only seem
to see cytoplasmic staining, and in general most tumors are showing
negative results.

I currently am using the Ultraview kit for optimization, our pathologist
likes CC2 for 44 min, Ab. Inc for 40 min using a 1:100 dilution.

Anyone have any suggestions that might help? If you want more detailed info
I can provide as well!

-- 

Erin McCarthy, HT (ASCP)
Pathology Lab Supervisor

Tempus Labs
600 W. Chicago Ave.
Chicago IL 60654
Ph:(312) 638-6344 Ext.3835

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Message: 3
Date: Thu, 18 Oct 2018 18:20:53 -0400
From: Eddie Martin <edmartin26 at gmail.com>
To: "Reilly, Laurie" <laurie.reilly at jcu.edu.au>
Cc: "Histonet at lists.utsouthwestern.edu"
	<Histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Restaining old Heamatology smears
Message-ID: <0E7F124E-9821-4A32-A438-63F45AD4566E at gmail.com>
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Hi Laurie,

Blood is considered a tissue so it is very much still Histo related. 

Regarding your smears, this also occurs in our hematopathology laboratory from time to time when we are scanning old slides and there?s a bubble under the coverslip that won?t let us take images with our digital scope. 

It may very well be that there is still a resinous permount film on your slides from the old coverslip. This may simply be removed by leaving the smear in xylene some additional time to remove the resin. Then place slides in 100% alcohol to remove xylene residues. The following steps would be rehydrating the slides with dilute changes in in alcohol...ie: 95%,then 85%, then 70%, then in a few changes of distilled water. If you can?t get the pH of your water to pH 7.2, then you could use a phosphate buffered solution to achieve this in place of the distilled water. 

Then place your slides in a working Giemsa working  solution. It may help to rock your slides or slightly agitate them to improve staining intensity. 

I hope this helps. 
Please feel free to contact me via email at Eddie.martin at NIH.gov

> On Oct 17, 2018, at 10:51 PM, Reilly, Laurie <laurie.reilly at jcu.edu.au> wrote:
> 
> Dear Histonetters,
> This is not quite Histo but related. One of our Laboratory Scientists has removed the coverslips from faded blood smears used for teaching and is having trouble restaining them with Wright's stain.
> Could we please call on the combined wisdom of the Histonet to try to find a solution to this problem.
> 
> Regards,      Laurie.
> 
> Mr. Laurie REILLY
> Histopathology
> Veterinary and Biomedical Sciences
> James Cook University
> Townsville  Qld.  4811
> Australia.
> 
> Phone 07 4781 4468
> Mobile 0448 957747
> 
> 

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Message: 4
Date: Thu, 18 Oct 2018 20:29:53 -0400
From: Eddie Martin <edmartin26 at gmail.com>
To: "Wilson, Lindsay A" <Lindsay.A.Wilson at uth.tmc.edu>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] BMP-2 protocol for bone
Message-ID: <1F29A020-476E-4627-8E40-A4F3F146A864 at gmail.com>
Content-Type: text/plain;	charset=utf-8

Hi Lindsay, 

There are a few complications  that will automatically come up due to working with rabbit tissue, and also due to the antibody from Abcam that you are using. 

I called Abcam prior to emailing you on a hunch that I had and was correct. The antibody you purchased from Abcam is unconjugated and first needs to be conjugated to whatever method you?re going to use. You could conjugate the antibody yourself. I already reached out to Abcam. It?s going to take them a few business days to get back to me to see whether their own antibody can be conjugated for IHC if that?s the route you?re going to go. 

Keeping this email simple for a response to your question, it may be easier to use another detection method for conjugating this specific antibody to rabbit bone...possibly FITC conjugated tyramide labeling with Alexa-Fluor. 

If you are insistent on using HRP it may be easier to contact me and we can further discuss as there are easily two complications that will arise due to the antibody you chose to optimize with and the species you are trying to stain on. 

Since you left your lab number on your post, I called and left my phone number on the voicemail for the young Lab if you would like to discuss and troubleshoot over the phone. I can also be reached via email at work at Eddie.martin at NIH.gov

Thanks,
Eddie

Eddie Martin HT(ASCP), QIHC(ASCP),HTL
Hematology Team Lead / Histology Technical Supervisor
The National Institutes of Health 
Clinic Center - Department of Laboratory Medicine
10 Center Drive, Room 2C 360
Bethesda, MD 20814

> On Oct 18, 2018, at 12:57 PM, Wilson, Lindsay A <Lindsay.A.Wilson at uth.tmc.edu> wrote:
> 
> Good morning,
> 
> Does anyone have a proven IHC protocol for BMP-2 on bone tissue? A big plus, if you have worked with it on rabbit bone!
> I am currently using Abcam?s BMP-2 (ab6285) with no success. I have performed antigen retrieval and non-antigen retrieval. I have used a dilution as low as 1:50. I appreciate any suggestions.
> 
> Lindsay Wilson, LVT, RLATG
> Young Laboratory
> 
> UTHealth | The University of Texas Health Science Center at Houston | School of Dentistry
> Department of Oral & Maxillofacial Surgery
> 
> 7500 Cambridge St. | Suite 6510 | Houston, TX 77054
> 713 486 4360 tel | 713 486 4333 fax
> 
> 

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