Highly Parallel Droplet Sample Preparation for Single Cell Proteomics v2

Protocol for preparing single cells for mass-spec analysis by nPOP as described by Leduc et al., 2021, 2022 DOI: 10.1101/2021.04.24.441211. nPOP uses piezo acoustic dispensing to isolate individual cells in 300 picoliter volumes and performs all subsequent preparation steps in small droplets on a fluorocarbon-coated slide. This design enables simultaneous sample preparation of thousands of single cells in a single batch. This includes lysing, digesting, and labeling individual cells in volumes below 20 nl. nPOP supports different experimental designs including label free analysis, TMT 18plex with carrier, mTRAQ, and TMT-10plex for TOF instruments (see steps). Data, resources and additional information: https://scp.slavovlab.net/Leduc_et_al_2022 and https://scp.slavovlab.net/nPOP Andrew Leduc presented a detailed workflow of nPOP at the 4th Single Cell Proteomics Conference available on YouTube: https://youtu.be/DJ1U_KpMNcY The protocol uses the CellenONE system for liquid handling and cell sorting and assumes basic level of familiarity and training on the CellenONE. If there are any questions, please email leduc.an@northeastern.edu