Table 1 Comparison of different single cell isolation methods
From: Microfluidics applications for high-throughput single cell sequencing
Methods | Description | Isolation process | Applicability | Throughput (cells per run) | Cost | Merits | Limitations |
|---|---|---|---|---|---|---|---|
Limiting dilution | Application of hand pipettes or pipetting robots to isolate single cells through dilution of the cell suspension | Manual/semi-automatic | Suspension cells | Low (< 100) | Low | Simple operation | Low specificity |
Low efficiency | |||||||
Low precision | |||||||
Cell loss | |||||||
Low work capacity (< 100) | |||||||
Micromanipulation | Application of inverted microscope combined with micropipettes to select and isolate single cells | Manual | Suspension cells | Low (< 100) | Low | Simple operation | Low efficiency |
Flexible sampling | Mechanical injury | ||||||
Visualized operation | High difficulty | ||||||
|  | Low work capacity (< 100) | ||||||
LCM | Application of infrared laser under a microscope to isolate single cell or cell compartments from solid tissue samples | Manual | Tissue samples | Low (< 100) | High | Maintain integrity of sample | Nuclear damage |
Genetic material loss | |||||||
RNA pollution | |||||||
High difficulty | |||||||
Low work capacity (< 100) | |||||||
FACS | Application of fluorescence labeling specific molecules on the cell surface to sort cells | Semi-automatic | Suspension cells | High (> 1000) | High | ·High specificity | Mechanical injury |
·High accuracy | Large sample amount | ||||||
·High sensitivity | Cannot process cells less than 1000 | ||||||
Traps-based microfluidics | Application of microfluidic chips to separate single cells through traps | Semi-automatic | Suspension cells | High (> 1000) | High | Flexible operation | Low specificity |
Efficient cell pairing and fusion | Partial stimulation on cells | ||||||
Valves-based microfluidics | Application of microfluidic chips to separate single cells through valves | Semi-automatic | Suspension cells | High (> 1000) | High | High sensitivity | Difficult and time-consuming fabrication |
High automation | Not portable | ||||||
Low sample volume | Â | ||||||
Droplet-based microfluidics | Application of microfluidic chips to separate single cells through droplets | Semi-automatic | Suspension cells | High (1000–10,000) | High | High sensitivity | Random encapsulation |
High specificity | Complex equipment | ||||||
Noise-free | Â |