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Cell hashing and sorting

SS Suzanne Isabelle Sitnikova
SM Sophie Munnings-Tomes
EG Elena Galvani
SK Stacy Kentner
KM Kathy Mulgrew
CR Chris Rands
JA Judit España Agustí
TZ Tianhui Zhang
KI Kristina M Ilieva
GR Guglielmo Rosignoli
HG Hormas Ghadially
MR Matthew J Robinson
TS Tim Slidel
RW Robert W Wilkinson
SD Simon J Dovedi
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FNA biopsies were collected from four mice bearing A20 tumors, a CD20+ B cell lymphoma model. The samples were stained with a mix of fluorescently labeled anti-CD20 PerCP/Cyanine 5.5 antibody (clone SA275A11) and TotalSeq-B0301 to B0304 anti-mouse Hashtag antibodies (a cocktail of DNA-barcoded anti-CD45 (clone 30-F11) and anti-MHC I clone (M1/42) antibodies in order to doubly label cells to increase tagging efficiency) (both Biolegend) according to the manufacturer’s protocol. The four hashtagged samples were pooled, stained with DRAQ7 viability dye (Abcam) and enriched for immune cells by sorting for CD20− cells on a BD FACSAria III cell sorter (BD Biosciences).

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