[Histonet] Immuno's

Mon Apr 11 10:46:11 CDT 2011

Toni beat me to it.

My suggestion is to contact the vendor where you purchase your detection.  The technical support departments for immuno vendors are emmensely helpful.  Try Dako at 800-400-DAKO, Biocare at 603-925-8000, or Cellmarque at 800.665.7284.  

In addition, it has always been my experience that a good hand stain is always better than an instrument stain -- primarily because they are better washed by hand.  My best guess is that you are not draining your slides well enough between rinses.  What I do is flood my slides with buffer, then put them at an angle (the Biocare IQ stainer is great for this) and rinse again -- Robin Simpkins at Biocare calls this the Jamaican waterfall.  I then follow up with a flick of the slide, then proceed to the next step.

Again call your vendors.  Technical support will be extremely helpful and you may get an expert (applications specialist or TSR) out to help you in person.

William Chappell, HTL(ASCP)QIHC
IHC Development Specialist
CellNetix Pathology & Laboratories
(503) 358-9567
www.cellnetix.com
"Our expertise...your peace of mind"

--- On Mon, 4/11/11, Rathborne, Toni <trathborne <@t> somerset-healthcare.com> wrote:

> From: Rathborne, Toni <trathborne <@t> somerset-healthcare.com>
> Subject: RE: [Histonet] Immuno's
> To: "Hannen, Valerie" <Valerie.Hannen <@t> parrishmed.com>, histonet <@t> lists.utsouthwestern.edu
> Date: Monday, April 11, 2011, 8:38 AM
> Who are you purchasing your
> antibodies from? Any of the companies that I have dealt with
> would gladly send someone out to optimize their antibodies.
> The fact that you are performing them manually probably is
> not the cause for the weak staining. We used to use the old
> Shandon Sequenza staining system as a backup if our Dako was
> out of service. When used with the same protocols, the
> staining was the same. 
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On
> Behalf Of Hannen,
> Valerie
> Sent: Monday, April 11, 2011 10:26 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Immuno's
> 
> 
> Hi Folks..
>  
> Looking for a bit of help. Is there anyone out there who
> are doing their
> immuno's manually?  Our Pathologist has just informed
> us that our
> immuno's are ALL too weak.
> He states that they always have been, as he compares them
> to slides that
> we get back from the  reference labs that we use for
> the antibodies that
> we don't do in-house.
>  
> If you are doing them manually...what companies are you
> using for
> antibodies, detection kit and chromogen? And are your
> stains really
> dark?
>  
> I informed our Pathologist that I know for a fact that the
> reference
> labs that we use, do the immuno's on stainers, and
> naturally these
> stains will be consistently be stronger.
>  
> I am trying to see if I can keep costs down by trying not
> to have to buy
> a stainer and continue to do the immuno's manually.
>  
> If you all know of a wet-workshop or of a immuno. company
> who would send
> someone to our facility to teach us ( the latter is
> preferred, since my
> section chief is going to retire in 1 1/2 weeks, I will be
> taking over
> her duties and this is going to leave us with only myself
> and one other
> tech, until we can get a replacement for my position).
>  
> Any help or advice that can be given will be greatly
> appreciated.
>  
> Thanks so much.
>  
> Valerie Hannen, MLT(ASCP),HTL,SU (FL)
> Parrish Medical Center
> Titusville,Florida
> 
> 
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