title: Automated analysis of dynamic behavior of single cells in picoliter droplets.
abstract: We present a droplet-based microfluidic platform to automatically track and characterize the behavior of single cells over time. This high-throughput assay allows encapsulation of single cells in micro-droplets and traps intact droplets in arrays of miniature wells on a PDMS-glass chip. Automated time-lapse fluorescence imaging and image analysis of the incubated droplets on the chip allows the determination of the viability of individual cells over time. In order to automatically track the droplets containing cells, we developed a simple method based on circular Hough transform to identify droplets in images and quantify the number of live and dead cells in each droplet. Here, we studied the viability of several hundred single isolated HEK293T cells over time and demonstrated a high survival rate of the encapsulated cells for up to 11 hours. The presented platform has a wide range of potential applications for single cell analysis, e.g. monitoring heterogeneity of drug action over time and rapidly assessing the transient behavior of single cells under various conditions and treatments in vitro.
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH - Royal Institute of Technology, Sweden. makho@kth.se.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Khorshidi
authors.forename: Prem Kumar Periyannan
authors.initials: PK
authors.lastname: Rajeswari
authors.forename: Carolina
authors.initials: C
authors.lastname: Wählby
authors.forename: Haakan N
authors.initials: HN
authors.lastname: Joensson
authors.forename: Helene
authors.initials: H
authors.lastname: Andersson Svahn
mesh_terms: D001331:Automation; D002470:Cell Survival; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal; D059010:Single-Cell Analysis / Q000379:methods*; D014867:Water / Q000737:chemistry
chemical_list: D004129:Dimethylpolysiloxanes; D014867:Water