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Single-cell sorting

KH Keren Bahar Halpern
HM Hassan Massalha
RZ Rachel K. Zwick
AM Andreas E. Moor
DC David Castillo-Azofeifa
MR Milena Rozenberg
LF Lydia Farack
AE Adi Egozi
DM Dan R. Miller
IA Inna Averbukh
YH Yotam Harnik
NW Noa Weinberg-Corem
FS Frederic J. de Sauvage
IA Ido Amit
OK Ophir D. Klein
MS Michal Shoshkes-Carmel
SI Shalev Itzkovitz
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Single cells were sorted with SORP-FACSAriaII machine (BD Biosciences) using a 100 μm nozzle. Dead cells were excluded on the basis of 500 ng/ml Dapi incorporation. Sorted cells were negative for CD45 and EPCAM and positive for PDGFRa. Cells were sorted into 384-well cell capture plates containing 2 μl of lysis solution and barcoded poly(T) reverse-transcription (RT) primers for single-cell RNA-seq28. Barcoded single cell capture plates were prepared with a Bravo automated liquid handling platform (Agilent) as described previously28. Four empty wells were kept in each 384-well plate as a no-cell control during data analysis. Immediately after sorting, each plate was spun down to ensure cell immersion into the lysis solution, snap frozen on dry ice and stored at −80 °C until processed.

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