Size-based sorting of hydrogel droplets using inertial microfluidics.

Hydrogel droplets encapsulating cells and molecules provide a unique platform in biochemistry, biology, and medicine, including single-cell and single-molecule analysis, directed molecular evolution, and detection of cellular secretions. The ability to prepare hydrogel droplets with high monodispersity can lead to synchronization of populations, more controlled biomaterials, and more quantitative assays. Here, we present an inertial microfluidic device for passive, continuous, and high-throughput sorting of hydrogel droplets by size. The sorting is achieved due to size-dependent lateral inertial equilibrium positions: hydrogel droplets of different sizes have different equilibrium positions under the combined effects of shear-gradient lift and wall-effect lift forces. We apply this separation technique to isolate smaller hydrogel droplets containing microalgal colonies from larger empty droplets. We found that hydrogel droplets containing microalga Euglena gracilis (E. gracilis) shrink as cells grow and divide, while empty hydrogel droplets retain their size. Cell-laden hydrogel droplets were collected with up to 93.6% purity, and enrichment factor up to 5.51. After sorting, we were able to recover cells from hydrogel droplets without significantly affecting cell viability.

[1]  K. Goda,et al.  A Gelatin Microdroplet Platform for High-Throughput Sorting of Hyperproducing Single-Cell-Derived Microalgal Clones. , 2018, Small.

[2]  Detlef Lohse,et al.  In-air microfluidics enables rapid fabrication of emulsions, suspensions, and 3D modular (bio)materials , 2018, Science Advances.

[3]  K. Goda,et al.  Shape-based separation of microalga Euglena gracilis using inertial microfluidics , 2017, Scientific Reports.

[4]  Zhi Zhu,et al.  Hydrogel Droplet Microfluidics for High-Throughput Single Molecule/Cell Analysis. , 2017, Accounts of chemical research.

[5]  Richard L. Brutchey,et al.  Flow invariant droplet formation for stable parallel microreactors , 2016, Nature Communications.

[6]  Dino Di Carlo,et al.  Accelerated wound healing by injectable microporous gel scaffolds assembled from annealed building blocks. , 2015, Nature materials.

[7]  Xavier Gidrol,et al.  Controlled 3D culture in Matrigel microbeads to analyze clonal acinar development. , 2015, Biomaterials.

[8]  Shigeru Shigeoka,et al.  Enhancement of photosynthetic capacity in Euglena gracilis by expression of cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase leads to increases in biomass and wax ester production , 2015, Biotechnology for Biofuels.

[9]  Yolanda Schaerli,et al.  Evolution of enzyme catalysts caged in biomimetic gel-shell beads. , 2014, Nature chemistry.

[10]  H. Amini,et al.  Inertial microfluidic physics. , 2014, Lab on a chip.

[11]  Hyung Jin Sung,et al.  Optical separation of droplets on a microfluidic platform , 2014 .

[12]  Wilhelm T S Huck,et al.  Probing cellular heterogeneity in cytokine-secreting immune cells using droplet-based microfluidics. , 2013, Lab on a chip.

[13]  H. Stone,et al.  Dripping and jetting in microfluidic multiphase flows applied to particle and fibre synthesis , 2013, Journal of physics D: Applied physics.

[14]  Abraham P Lee,et al.  Passive droplet sorting using viscoelastic flow focusing. , 2013, Lab on a chip.

[15]  Helene Andersson Svahn,et al.  Droplet microfluidics--a tool for single-cell analysis. , 2012, Angewandte Chemie.

[16]  Haakan N Joensson,et al.  Droplet size based separation by deterministic lateral displacement-separating droplets by cell--induced shrinking. , 2011, Lab on a chip.

[17]  Nicole K Henderson-Maclennan,et al.  Deformability-based cell classification and enrichment using inertial microfluidics. , 2011, Lab on a chip.

[18]  J. Shuga,et al.  Single-cell multiplex gene detection and sequencing with microfluidically generated agarose emulsions. , 2011, Angewandte Chemie.

[19]  Tae Seok Sim,et al.  Multistage-multiorifice flow fractionation (MS-MOFF): continuous size-based separation of microspheres using multiple series of contraction/expansion microchannels. , 2011, Lab on a chip.

[20]  Chaoyong James Yang,et al.  Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR. , 2010, Lab on a chip.

[21]  D. Di Carlo,et al.  Continuous scalable blood filtration device using inertial microfluidics , 2010, Biotechnology and bioengineering.

[22]  H. Amini,et al.  Label-free cell separation and sorting in microfluidic systems , 2010, Analytical and bioanalytical chemistry.

[23]  A. Abate,et al.  Ultrahigh-throughput screening in drop-based microfluidics for directed evolution , 2010, Proceedings of the National Academy of Sciences.

[24]  D. Di Carlo,et al.  Sheathless inertial cell ordering for extreme throughput flow cytometry. , 2010, Lab on a chip.

[25]  X. Mu,et al.  On-chip manipulation of continuous picoliter-volume superparamagnetic droplets using a magnetic force. , 2009, Lab on a chip.

[26]  A. Abate,et al.  Surface acoustic wave (SAW) directed droplet flow in microfluidics for PDMS devices. , 2009, Lab on a chip.

[27]  Christoph A. Merten,et al.  Drop-based microfluidic devices for encapsulation of single cells. , 2008, Lab on a chip.

[28]  M. Yamada,et al.  Continuous and size-dependent sorting of emulsion droplets using hydrodynamics in pinched microchannels. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[29]  R. Tompkins,et al.  Continuous inertial focusing, ordering, and separation of particles in microchannels , 2007, Proceedings of the National Academy of Sciences.

[30]  Helen Song,et al.  Reactions in droplets in microfluidic channels. , 2006, Angewandte Chemie.

[31]  A. Khademhosseini,et al.  Hydrogels in Biology and Medicine: From Molecular Principles to Bionanotechnology , 2006 .

[32]  S. Hutner,et al.  High yield media for photosynthesizing euglena gracilis abstract , 1967 .

[33]  M. Demirbas,et al.  IMPORTANCE OF ALGAE OIL AS A SOURCE OF BIODIESEL , 2011 .