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Table 1 Single-cell functional proteomics tools

From: Microchip platforms for multiplex single-cell functional proteomics with applications to immunology and cancer research

Technique Numbers and types of poteins assayed Through-put Detection limit Statistical accuracy and signal quantification Notes and features Literature
Flow cytometry methods
Fluorescence flow cytometry Around 15 proteins (mostly membrane proteins, a few cytoplasmic proteins) 104 cells s-1 500 copies per cell 90% phenotyping accuracy; relative protein abundance Standard for sorting and enumeration of cellular phenotypes. Secretion blocked and cells fixed for cytoplasmic proteins [5, 6]
Mass flow cytometry Around 35 membrane and intracellular proteins, likely expandable 103 cells s-1 >103 copies per cell Good cell counting statistics; relative protein abundance Cells handled in bulk prior to analysis. Secretion blocked and cells fixed for cytoplasmic proteins [8]
Surface methods
ELISpot 1-3 secreted proteins   6 spots per 105 cells Quantitative for percentage active cells Cells secrete proteins onto antibody coated surfaces; secretion activity correlated with cell location [9]
Microfluidics technologies
Image cytometry 3-4 membrane or intracellular proteins and cell size 103-104 cells per chip 105 fluoro-phores per μm2 Good cell counting statistics; relative protein abundance Cells are fixed and stained (in bulk) with fluorescent antibodies; protein assay and cell location spatially correlated [19, 20]
Cell array 1 intracellular protein <103 cells per chip   Good cell counting statistics; relative protein abundance Single cells separated and imaged on chip; continuous monitoring of cell physiology [12–15]
Micro-droplet 1 membrane or intracellular protein 102 μdrops s-1 Not defined Good cell sampling statistics Cells entrained in microdroplets; microdroplet composition control permits screening cells [16–18]
Micro-engraving 3 secreted or 3 membrane proteins 104-105 cells per chip Not available Very good cell number statistics; relative protein abundance Cells isolated in miocrowells; surface immunoassays; proteins colorimetrically detected; secretome kinetics from single cells; proteomeic and functional assays from same cell [21–23, 37]
Single cell barcode chips About 20 secreted, membrane, or cytoplasmic proteins, expandable 103-105 cells per chip 102 copies Good cell counting statistics, absolute quantification, 10% measurement error per protein per cell Cells isolated in microchambers, miniature antibody arrays yield spatial separation of specific protein assays; proteomeic and functional assays from same cell; single cells or defined small cell populations accessed. [4, 5, 26–30]
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Genome Medicine

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