[Histonet] FITC on Ventana Ultra

Mark Tarango marktarango <@t> gmail.com
Fri Aug 27 12:18:07 CDT 2010


Hi Nita,

I agree that buffer should be used as the negative control reagent.  If you
have a seperate slide that is cut and put into buffer, just coverslip it
with the same mounting media and you have a negative control.  All the
instrument is doing is putting on the antibody and then rinsing it off.  So
your negative control is a slide that didn't get the antibody (just
buffer).  Makes sense to me.  If you get ventana to add a negative control
to their software it would do just the same thing as coverslipping from
buffer, but you'd be paying ventana for it.

Mark
On Fri, Aug 27, 2010 at 7:30 AM, Nita Searcy <NSEARCY <@t> swmail.sw.org> wrote:

> Any users out there that have had A CAP inspector question negative control
> on the instrument? In regard to CAP question ANP.21850 in which the notes
> state, "A negative reagent control in which the patient tissue is processed
> in an identical manner to the test specimen but with the primary antibody
> omitted must be performed for each patient test specimen?"
>
> Below is the response from Ventana regarding "negatives" I am curios if CAP
> accepts these methods??
>
> In regards to the question below about running a negative control fitc,
> there are two ways to accomplish this.
>
>
>
> 1) There is not currently a place in the protocol to select a fitc
> negative. We can still be compliant by ensuring the negative slide is
> treated the same as the patient. The only reagent the patient slide is
> exposed to, besides the fitc antibody, is reaction buffer. Running a
> negative can be accomplished by applying reaction buffer to the slide and
> letting it incubate for the same amount of time. Next, it is important to
> ensure both slides are coverslipped in the came mounting media.
> 2) The second method is to utilize internal negatives that are already
> present within the patient tissue. However, this is difficult when running
> Fitc antibodies such as IgG, IgM, etc.
> We are looking into adding this addition into the software. Many customers,
> however have expressed that they would rather utilize the slide drawer for
> another patient slide, rather than running the fitc negative.
>
> Am interested in your comments.
> Thanks
>
>
>
>
>
> Nita Searcy, HT/HTL (ASCP)
> Scott and White Hospital
> Division Manager, Anatomic Pathology
> 2401 S. 31st. Street
> 254-724-2438
> Temple, Texas, 76502
> nsearcy <@t> swmail.sw.org
>
>
> 254-724-2438
>
>
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