Ventana Staining

Does this antibody work? Designing a new panel?
Antibodies, isotopes, etc. for Imaging Mass Cytometry applications
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jfathman
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Joined: Wed Jan 10, 2018 4:09 pm

Ventana Staining

Post by jfathman » Wed Jan 17, 2018 4:26 pm

Does anyone use one of the Ventana machines to stain their sections for IMC? We have the Discovery XT and the Ultra and are planning on testing them out in our workflow. Quick question, normally we would stain overnight at 4 degrees, but my histology core is telling me I only need to stain for 1hr with the Ventana, due to the small volume and liquid coverslip. Any experience or advice with this?
Thanks
-Jake
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bernd.bodenmiller
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Re: Ventana Staining

Post by bernd.bodenmiller » Fri Jan 19, 2018 8:57 am

We do not have experience with the Ventana machines, but Hartland Jackson in my lab compared O/N@4°C with 1h@RT antibody staining and he got much more background with the 1h@RT antibody staining. Thus we prefer the O/N@4°C condition. But who knows, things might be different with the Ventana set-up and only an experiment will provide the answer.
drimm
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Re: Ventana Staining

Post by drimm » Sat Jan 20, 2018 8:11 am

While we do not have experience with the Ventana stainer for IMC or heavy metal conjugated antibodies, we did some work a few years ago with a Ventana stainer and Cy5 tyramide. We found that the standard Cy5 tyramide was incompatible with the liquid coverslip, possibly partitioning in the oil phase or perhaps altered in some other way. Although we never figured it out, the Cy5 tyramide was incompatible with the Ventana stainer. Thus appropriate controls should be used when testing new labeled antibodies in the context of the liquid coverslip.
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Jkburks
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Re: Ventana Staining

Post by Jkburks » Thu Feb 22, 2018 3:29 pm

I am very curious about the Ultra as I have a researcher who has used his to get excellent 7 color labeling for the Opal system. He did not use the liquid coverslips mentioned by David. Also of note is that his Ventana platform is completely open (meaning he has full control of every step in the process) and I don't know if that is common or not. We have a heavy user base for the Vectra and building a heavy list for the Hyperion, so I am also debating a stainer and the Ventana is leading my list as it would be nice way to standardize a pipeline.

David, for the longer wavelength in Opal/TSA Prolong Diamond Mounting media has been shown to increase the signal. Not sure anyone knows why, but if you are still working with it you should give that a try.

Jared
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