[Histonet] RE: Sequenza units for manual immunohistochemistry (Keri Colwell)

[Liebig, Tyler K.]

Hello,

This is Tyler Liebig with the Thermo Scientific IHC group.  I'm responding to 
Keri's questions regarding the Sequenza manual staining.  Keri you are correct 
that if the waste is not emptied and builds up too much in the bottom of the 
sequenza unit it will definitely stop the capillary action (Reagent can't flow 
both ways :-).  However, this can be avoided by emptying the waste regularly 
between runs.  

You bring up a good question though that I didn't know the answer to: "How many 
slides can be stained before the waste hits the slides?" I did a quick 
calculation below that I hope is helpful.  

Total Waste Volume of Sequenza Rack:
In a Sequenza rack the slides are held about 1.5 inches above the bottom of the 
tray.  This allows about 270ml of waste before the level is close touching the 
slides.  

Reagent use per slide:  If you stain a single slide with a long protocol then 
worst case scenario is about 7 steps.  (H2O2 through Counterstain with a two 
step polymer)
Volume of rinse buffer: This means you would rinse 8 times 2ml each (16ml total 
rinse buffer per slide worst case).
For the staining reagents: The recommended volume is 0.1ml per reagent and 
0.5ml for chromogen.  So for 6 reagents plus chromogen that is 1.1ml and I will 
double it because I know some people add way more than needed.  So 2.2ml per 
slide worst case.

So to sum it up worst case scenario 18.2ml of waste is created per slide which 
means 14 slides (270/ml/18.2ml=14.8) can be stained before the waste risks 
touching the slides.  The rack holds ten slides total. 

In the end I recommend users play is save and empty the waste after each batch 
of slides stained even though most user generate much less waste than the 
calculations above.

I also included a link to a great description of Sequenza use on the Histonet 
posted by Gayle M Callis HTL/HT/MT (ASCP).  I have referenced these details 
frequently myself (Thanks Gayle).
http://www.histosearch.com/histonet/Jul02A/Longansweronusetechnicswi.html

Hope that helps and if you have any other questions I'm happy to help offline 
(contact information is below).

Thank you,

Tyler Liebig
Americas Product Manager - Immunohistochemistry
Thermo Scientific Anatomical Pathology

 (510) 979-5000 ex 31816
(510) 299-1751 mobile

tyler.lie...@thermofisher.com

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[Histonet] RE: Histonet Digest, Vol 78, Issue 2

[Liebig, Tyler K.]

agents and
can be modified to your facility and reagent list.

Keith M. Mikoff, HTL/HT (ASCP)

  PAPANICOLAOU STAIN (PAP)

  PURPOSE: The following staining procedure is to be performed only
  on spray fixed slides.  It is a routine stain used for any cytol-
  ogy (i.e., PAP smears, FNA procedures, or specimens of a fluid
  nature where cytology is required).

  PROCEDURE:

  1.   Dehydrate slides in 2 changes of isopropyl alcohol, 5
  minutes each.

  2.   Rinse in tap water for two minutes.

  3.   Stain in Harris' Hematoxylin for 2 minutes.

  4.   Rinse in fresh tap water for 3 minutes.

  5.   Place in ammonia water for 1 minute.

  6.   Rinse in fresh tap water for 1 minute.

  7.   Dehydrate in 3 changes of Isopropyl alcohol, 2 minutes each.

  8.   Stain in O-G 6 for one minute.

  9.   Dehydrate in Isopropyl alcohol for 2 changes, 2 minutes
  each.

  10.  Stain in EA-50 for 4 minutes.

  11.  Place in Isopropyl alcohol, 3 changes, at 1 1/2 minutes each

  12.  To clear the slides, place in 3 changes of Histoclear, for 2
  minutes each, and coverslip.









--

Message: 4
Date: Sun, 2 May 2010 09:01:04 -0400
From: 
Subject: RE: [Histonet] disposable blade holder
To: 
Message-ID: <1729e60b0b6d456bb67e686c815cc...@hopperpc>
Content-Type: text/plain;   charset="US-ASCII"

As a comical sidelight to the last sentence that Tyler wrote ...

I have a pathologist who was very upset that our cryostat didn't seem to be
working properly.  I rushed up to the frozen room to assist and he said,
"well, nevermind, I got a slide and diagnosis."  I looked in the cryostat
and said to him "you actually got a section and were able to render a
diagnosis?"  "yes" he said.  "WOW!, you must be the ONLY pathologist who is
capable of sectioning tissue without a knife ..."  ;o)  He sectioned the
tissue and truly had no idea there wasn't a knife in the blade holder!

:o)



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liebig,
Tyler K.
Sent: Friday, April 30, 2010 3:41 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] disposable blade holder


Hello Lucy,

In regards to your trouble finding an HM310 blade holder we do have one in
our refurbished pool that I could have sent to you. It is used but in
working order so we can let you have it at no charge.  If you let me know
where to ship it I will have it sent.

Hope that helps, It's hard to cut without a blade holder.

Regards,

Tyler Liebig
Product Manager
Histology Instrumentation and IHC
Anatomical Pathology
Thermo Fisher Scientific
46360 Fremont Blvd
Fremont, CA 94538
USA
Office: (510) 979-5000
Mobile: (510) 299-1751
Fax: (510) 979-5239
tyler.lie...@thermofisher.com<mailto:tyler.lie...@thermofisher.com>
www.thermo.com<http://www.thermo.com>


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Message: 5
Date: Sun, 2 May 2010 09:04:16 -0400
From: 
Subject: RE: [Histonet] Slide and Block Storage
To: 
Message-ID: <7180df002734414d8feee9a4a4600...@hopperpc>
Content-Type: text/plain;   charset="US-ASCII"

In our facility, we maintain both the blocks and slides.

We also store older blocks/slides in a climate controled an off-site
facility.


Michelle


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth
Kronenberger
Sent: Friday, April 30, 2010 11:00 AM
To: 'Nails, Felton'; 'Daniel Adams'; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Slide and Block Storage


Clients fax orders for deepers and special stains which we perform and send
them out with their next package for them to read and report.

-Original Message-
From: Nails, Felton [mailto:flna...@texaschildrens.org] 
Sent: Friday, April 30, 2010 10:48 AM
To: 'Elizabeth Kronenberger'; 'Daniel Adams';
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Slide and Block Storage

So what if the reporting company need to order special stains, it adds too
much time having to request the blocks or additional unstained slides. 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth
Kronenberger
Sent: Friday, April 30, 2010 9:40 AM
To: 'Daniel Adams'; histonet@lists.utsouth

[Histonet] disposable blade holder

[Liebig, Tyler K.]

Hello Lucy,

In regards to your trouble finding an HM310 blade holder we do have one in our 
refurbished pool that I could have sent to you.
It is used but in working order so we can let you have it at no charge.  If you 
let me know where to ship it I will have it sent.

Hope that helps, It's hard to cut without a blade holder.

Regards,

Tyler Liebig
Product Manager
Histology Instrumentation and IHC
Anatomical Pathology
Thermo Fisher Scientific
46360 Fremont Blvd
Fremont, CA 94538
USA
Office: (510) 979-5000
Mobile: (510) 299-1751
Fax: (510) 979-5239
tyler.lie...@thermofisher.com
www.thermo.com


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