Table 1.
Table summarizing different single-cell analysis tools discussed in this review in terms of their advantages, disadvantages, and applications.
| Research applications | Advantages | Limitation | Commercial vendors | References | |
|---|---|---|---|---|---|
| Cytometry | Cytokine and surface markers Signaling and activation dynamics Cytotoxicity Immunophenotyping Sorting |
High throughput Quick to run (on the order of several hours) Ability to sort cells based on expression of molecules |
Spectral overlaps Expensive Not possible to replicate dynamic cellular microenvironment Static snapshot over time |
BD Biosciences, MilliporeSigma, Miltenyi Biotec, Thermo Fisher Scientific | (177), (178), (179) |
| MICROSYSTEMS | |||||
| Nano wells | Antibody and drug screening Activation dynamics Cytotoxicity Inter and intra cellular communication Cytokines and signaling molecules release |
High-throughput Possibility to recover cells from the wells Simple to use Multiplexing and real time monitoring Quantitative analysis of proteins Possibility to pair multiple cells in each well |
Seeding cells under the effect of gravity lowers the efficiency of cell loading Manual operation Limited control over the fluidic and cellular microenvironment Random distribution of cells in wells |
μFluidix, microfluidic ChipShop | (76), (77), (79), (80), (163), (55), (78) |
| MICROFLUIDICS | |||||
| Trap-Based | Inter and intracellular communication Cytotoxicity Signaling and activation dynamics Cytokines and signaling molecules release |
Efficient cell pairing and fusion Ability to monitor cellular behavior, from the point of cellular interaction and contact Multiplexing and real time monitoring High throughput |
Stimulation of cells in bulk and not on-chip (Juxtacrine and paracrine interactions before loading the cells cannot be ruled out) Controlled on-chip stimulation is not possible Cells are not isolated from each other (The trapped cells and secreted molecules are not completely confined and this can influence the behavior of the neighboring cells by paracrine communication) |
μFluidix, microfluidic ChipShop | (104), (105), (106), (96), (109) |
| Valve-based | Signaling and activation dynamics Cytokines and signaling molecules release Monitor transcription factors activity Migration |
Complete automation Replication of dynamic cellular microenvironment Sensitive (addressable cell chambers with very small volume) Low sample volume Both real time monitoring and end-point analysis |
Laborious and time-consuming fabrication Not portable (additional equipment for operation) Low to medium throughput |
Fluidigm C1 | (128), (129), (135), (16), (180), (181), (182) |
| Droplet-based | Inter and intracellular communication Cytokines and signaling molecules release Activation dynamics Antibody screening Cytotoxicity |
High throughput Noise-free cellular microenvironment due to compartmentalization Both real time monitoring and end point analysis Multiple as well as single cell pairing Sensitive (low reaction volume) Small sample and reagent volume |
Not possible to replicate dynamic cellular microenvironment Random encapsulation (follows Poisson Distribution) Difficult to incorporate washing steps |
μFluidix, microfluidic ChipShop, Dolomite, Fluigent, 10X Genomics (Chromium Controller) BIORAD (ddSEQ) | (144), (162), (164), (166), (167), (161) |
The applications described here are what is presented in this review as well as all the other potential applications of the design for immune cell analysis at single-cell level.