Democratized high-throughput single-cell secretion screening using droplets formed by structured microparticles
Joe de Rutte, Robert Dimatteo, Mark van Zee, Robert Damoiseaux, and Dino Di Carlo
Monday October, 28 12:00
Session 1A2 San Francisco Room
We demonstrate a lab-on-a-particle approach to analyze and sort single cells based on secretions without the use of specialized equipment. Shaped microparticles hold cells and form uniform sub-nanoliter water-in-oil compartments upon simple agitation. Single cell secretions are captured on the particles and the associated cells are sorted using commercial flow sorters. Because we avoid microfluidics in the workflow, this lab-on-a-particle approach should can be easily adopted by non-specialist researchers.
Vishwesh Shah*, Yilian Wang*, Joseph de Rutte, Chueh-Yu Wu, and Dino Di Carlo
Microgel Templated Droplet ELISA
Tuesday 29, October 17:00
Session 2B3 Singapore Room
We present amplified affinity assays performed on monodisperse microgel particles, which template uniform droplets without using microfluidics. The signal from the assay is immobilized on each particle, and therefore can be quantified using standard flow cytometers. This lab-on-a-particle platform can democratize ultra-sensitive immunoassays, advancing the discovery of extremely rare biomarkers, as well as redefining disease diagnosis.
Wednesday October, 30 14:15 - 16:45
Here we apply magnetic ratcheting to sort clonal cell populations based on the amount of magnetic biomass, magnetosome, accumulated in uniformly sized gelatin microgels, towards the directed evolution of new highly productive strains of magnetic particle-producing bacteria. Magnetic ratcheting system is the versatile sorting tool based on target magnetic contents. We have showed the magnetic ratcheting system can sort out 1.55 times higher magnetosome production population than non-selected population. As the next step, we will do multiple selection cycle with chemical mutagenesis to produce super productive magnetotactic bacteria population.
Magnetic ratcheting of hydrogel drops for selection of high magnetic biomass production bacteria
Hiromi Miwa, Hayley McCausland, Coleman Murray, Arash Komeili, and Dino Di Carlo
Droplet-Enhanced On-Cell Encoding of Single Cell Secretory Function
Robert Dimatteo and Dino Di Carlo
Monday October, 28 14:00 - 16:30
Engineered T-cells have demonstrated profound promise towards the treatment of otherwise intractable cancers. However, the inherent heterogeneity of individual clones in cell products is often neglected, limiting reproducibility. We report a method of sorting single viable T-cells directly through secreted protein signatures using conventional flow cytometry. T-cells are rapidly encapsulated into 100 µm droplets with cell-linked anticytokine capture and detection antibodies, prompting the formation of fluorescent sandwich immunocomplexes on the surface of secreting clones. Partitioning samples enables continued signal accumulation without intercellular crosstalk, enhancing the potential to probe secreted proteins over a large dynamic range.