[Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

[Vanessa Perez]

You just need the FITC antibodies.  No kit required.  The protocol is under the procedure Fluorescence IHC
Basically all the machine does is dispense the antibody on the slide for however long you choose.
And some rinses of reaction buffer...

Vanessa 

-----Original Message-----
From: Truscott, Tom [mailto:ttruscot <@t> vetmed.wsu.edu] 
Sent: Monday, April 22, 2013 3:52 PM
To: Vanessa Perez
Subject: RE: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

Hi Vanessa, I've been following this line of email, since I may need to start doing fluorescence on XT. Which kit/program/protocol do you use on the XT? Can you run them without a kit? Thankyou, Tom T

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Vanessa Perez
Sent: Monday, April 22, 2013 1:22 PM
To: elonergan <@t> metrocast.net; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

Yes, well when we take the ones off of the XT we add the neg and do the rinsing steps all together.
We do two rinses of reaction buffer for 5 min then final rinse with DI water...then coverslips....



Vanessa 

-----Original Message-----
From: elonergan <@t> metrocast.net [mailto:elonergan <@t> metrocast.net] 
Sent: Monday, April 22, 2013 3:10 PM
To: Vanessa Perez
Subject: Re: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

Vanessa,  we are just getting our DIF up and running. For your neg, it doesn't go thru any step, just coverslip direct from buffer?  Thanks Eileen Lonergan MGH dermatopath associates.  
Sent via BlackBerry by AT&T

-----Original Message-----
From: Vanessa Perez <vperez <@t> pathreflab.com>
Sender: histonet-bounces <@t> lists.utsouthwestern.edu
Date: Mon, 22 Apr 2013 14:40:39 
To: histonet <@t> lists.utsouthwestern.edu<histonet <@t> lists.utsouthwestern.edu>
Subject: RE: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

We use Benchmark here with no problems.  We use tonsil controls to validate it here.  For the neg. we just have an extra slide sit in reaction buffer while the other slides are on the machine.
We switched from manual b/c less cost and less tech time.



Vanessa 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sue Hunter
Sent: Monday, April 22, 2013 2:30 PM
To: Mel John del Barrio; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

We run our DIF on our old Lab Vision stainer because it is easier and cheaper.  The Ultra ready to use Abs are very expensive and there is no negative control.  As far as we can find out, there is no prep kit that you can use with the fluorescence protocols to make your own.  Ventana's solution is to put your negative slide in a coplin jar filled with buffer and then add that to your slide tray when you are done.
We have the same issue with our Bond Max stainer - you have to use a detection kit for everything on the Bond so Leica's solution is to sell you a kit really cheap where you only use one solution and  throw the rest away. Very expensive.
Since neither one of these "solutions" is an acceptable one for us, we still do them the "old"way and are very happy. I would think that unless you are running a really large number of cases, even doing them by hand is preferable to running them on the Ultra.
Just my opinion.....
Sue


Sue Hunter, Supervisor
Advanced Diagnostics
Beaumont Health System
Royal Oak MI
248-898-5146
shunter <@t> beaumont.edu






-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mel John del Barrio
Sent: Monday, April 22, 2013 12:49 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Direct Immunofluorescence on Benchmark XT/Ultra

Hi  All
 
Anyone in the group utilises the Benchmark for DIF's? What sort of controls do you use to validate the assay ? What problems  you have encountered?
 
 
Thanks
 
MJ
 
 


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