Summary of microfluidic technologies for vesicle preparation.

Preparation methods and references	Advantages	Disadvantages
Microfluidic hydrodynamic focusing (MHF):15,68–75,77–84,86–90	High-throughput and continuous production of nano-sized vesicles	Residual organic solvent
	Monodisperse vesicle size and lamellarity	Limited range of size and lamellarity
	High encapsulation efficiency	Low production concentration
Micromixer	High-throughput and continuous production of nano-sized vesicles	Residual organic solvent
Staggered herringbone mixer (SHM):45,103,107–116	High encapsulation efficiency	Less monodisperse size compared with MHF
Twisted channel (iLiNP):119,120	Highly commercialized
Dean flow:101,122,124–129
On-chip hydration 134–136	Oil-free	Batch production
	Homogeneous size controlled by microstructures	Polydisperse lamellarity
		Low encapsulation efficiency
Microfluidic refined emulsion phase transfer 145–147	High-throughput and continuous generation of droplets	Low-throughput batch formation of vesicles
	Monodisperse size and lamellarity	Residual oil solvent
	High encapsulation efficiency	Bursting of droplets
	Generation of membrane asymmetry
Ice droplet hydration 149,150	Clean removal of organic phase	Low encapsulation efficiency
	Monodisperse vesicle size	Uncontrollable lamellarity
	Avoiding extensive droplet coalescence
Microfluidic single emulsion transfer 151–155	Continuous production of cell-sized vesicles	Low-throughput vesicle formation
	High-throughput generation of droplets	Residual oil solvent
	Monodisperse size and lamellarity	Bursting of droplets
	High encapsulation efficiency
	Adaptability for asymmetric vesicles
Microfluidic double emulsion-based vesicle generation 138,163,166–170,173,174,177–180	High-throughput and continuous production of cell-sized vesicles	Residual oil solvent
	Monodisperse size and lamellarity	Sophisticated equipment
	High encapsulation efficiency
	Adaptability for asymmetric vesicles, multi-compartmental vesicles
Continuous droplet interface cross encapsulation (cDICE) 181–188	Easy fabrication and affordable device cost	Limited oil selection
	Uniform size high encapsulation efficiency	Poor partition of specific lipids
	Adaptability for asymmetric vesicles
Pulsed jetting 46,190–200	High-throughput production of cell-sized vesicles	Residual oil solvent
	High encapsulation efficiency	Existence of small satellite vesicles
	Adaptability for asymmetric vesicles, multi-compartmental vesicles and nano-sized vesicles
On-chip electroformation 201–204	Oil-free	Batch production
	Highly unilamellar population	Hard asymmetry construction
	Good encapsulation efficiency	Hard protein incorporation