Table 1.

Comparison of different single cell isolation methods

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
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

LCM Laser capture microdissection, FACS fluorescence activated cell sorters