The Lab Notebook

Wushouer Ouerkaxi, PhD, MD

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Gating strategy for tetramer analysis to eliminate non-specific event

Posted by Wushouer Ouerkaxi, PhD, MD on Nov 14, 2018 1:00:00 PM

While MHC tetramer staining of antigen specific T cells for flow cytometry is considered highly specific, it could still be associated with non-specific staining. The gating strategy shown below is an example that investigators have used to reduce non-specific staining events using a dump channel. A dump channel can be planned on a case by case basis, on the experimental design, and consists of a pool of antibodies against cells that do not express our specific targets. For example, CD8+ T cells, anti-CD4/anti-CD14/anti-CD19 may be used to eliminate these immune cells from our targeted gate, and all may be labeled with the same fluorochrome. 

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Tags: Tetramer, Flow Cytometry

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