Droplet proteomic sample preparation (nPOP)
The nano-ProteOmic sample Preparation (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, including lysing, digesting, and labeling individual cells in volumes below 20 nl.
Advantages of nPOP
- nPOP can prepare thousands of single cells in a single batch
- This minimizes the # of batches and associated technical variation
- nPOP uses only mass-spec compatible chemicals and volumes below 20 nl
- nPOP droplet layouts are computationally programmable and thus very flexible
- Thus nPOP can be used for all multiplexed or label-free workflows without specialized consumables
- nPOP uses accessible consumables and small amounts of labels
- This keeps costs down to about 10-20 cents / single cell.
nPOP uses the CellenONE instrument, which is commercially available. Without access to CellenONE, one may use mPOP, which can prepares a few hundred single cells in parallel. Both mPOP and nPOP are fully compatible with all mass-spectrometry methods described in this portal.
Applications & data
- Leduc et al, 2021 used nPOP to prepare samples for SCopE2 analysis of the cell division cycle of U937 and Hela cells. Leduc et al, 2021 data website
- Derks et al, 2022 used nPOP to prepare samples for plexDIA analysis of single melanoma, monocyte and pancreatic adenocarcinoma cells. Derks et al, 2022 data website
- Leduc et al, 2022 used nPOP to prepare samples for pSCopE analysis of protein covariation in monocytes and melanoma cells. Leduc et al, 2022 data website
- Huffman et al, 2022 used nPOP to prepare samples for pSCopE analysis of primary macrophages stimulated with LPS. Huffman et al, 2022 data website
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