Prestained cells on different CyTOF models

Comparing performance of CyTOFv1, CyTOFv2, CyTOFv2-to-Helios upgrade, and two Helios instruments on aliquots of frozen prestained PBMC samples. PBMC samples from pooled vials of same donor, stained with surface antibodies, fixed with 2% PFA, permeabilized with saponin, stained with Ir intercalator, pooled again, and resplit into replicate aliquots that were then frozen at -80C in FBS+10% DMSO until use. Therefore, the samples are as identical as possible, leaving instrumentation as the main variable. All files are in Di mode. Compared Raw (no normalization) vs Fluidigm-ver2 normalization vs MATLAB normalization for each instrument. v2 instrument was upgraded to create the v2-to-Helios instrument. Notes: Freezing as 2016-Sumatoh et al-Cytometry A (PMID: 27798817) Staining as 2015-Leipold et al-Methods Mol. Biol (PMID: 26420710) Panel of antibodies+metals in Attachments section Excel