Many important biological questions demand single-cell transcriptomics on a large scale. Hence, new tools are urgently needed for efficient, inexpensive manipulation of RNA from individual cells. We report a simple platform for trapping single-cell lysates in sealed, picoliter microwells capable of printing RNA on glass or capturing RNA on beads. We then develop a scalable technology for genome-wide, single-cell RNA-Seq. Our device generates pooled libraries from hundreds of individual cells with consumable costs of $0.10–$0.20 per cell and includes five lanes for simultaneous experiments. We anticipate that this system will serve as a general platform for single-cell imaging and sequencing.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0684-3) contains supplementary material, which is available to authorized users.
Controlled transport of multiple individual nanostructures is crucial for nanoassembly and nanodelivery but is challenging because of small particle size. Although atomic force microscopy and optical and magnetic tweezers can control single particles, it is extremely difficult to scale these technologies for multiple structures. Here, we demonstrate a "nano-conveyer-belt" technology that permits simultaneous transport and tracking of multiple individual nanospecies in a selected direction. The technology consists of two components: nanocontainers, which encapsulate the nanomaterials transported, and nanoconveyer arrays, which use magnetic force to manipulate individual or aggregate nanocontainers. This technology is extremely versatile. For example, nanocontainers encapsulate quantum dots or rods and superparamagnetic iron oxide nanoparticles in <100 nm nanocontainers, the smallest magnetic composites to have been simultaneously moved and optically tracked. Similarly, the nanoconveyers consist of patterned microdisks or zigzag nanowires, whose dimensions can be controlled through micropatterning. The nanoconveyer belt technology could impact multiple fields, including nanoassembly, biomechanics, nanomedicine, and nanofluidics.
We present a multiplex method, based on microscopic programmable magnetic traps in zigzag wires patterned on a platform, to simultaneously apply directed forces on multiple fluid-borne cells or biologically inert magnetic micro-/nano-particles. The gentle tunable forces do not produce damage and retain cell viability. The technique is demonstrated with T-lymphocyte cells remotely manipulated (a la joystick) along desired trajectories on a silicon surface with average speeds up to 20 μm/s.
A platform of discrete microscopic magnetic elements patterned on a surface offers dynamic control over the motion of fluid-borne cells by reprogramming the magnetization within the magnetic bits. T-lymphocyte cells tethered to magnetic microspheres and untethered leukemia cells are remotely manipulated and guided along desired trajectories on a silicon surface by directed forces with average speeds up to 20 microm/s. In addition to navigating cells, the microspheres can be operated from a distance to push biological and inert entities and act as local probes in fluidic environments.
The demand for high-throughput single cell assays is gaining importance because of the heterogeneity of many cell suspensions, even after significant initial sorting. These suspensions may display cell-to-cell variability at the gene expression level that could impact single cell functional genomics, cancer, stem-cell research and drug screening. The on-chip monitoring of individual cells in an isolated environment would prevent cross-contamination, provide high recovery yield, and enable study of biological traits at a single cell level. These advantages of on-chip biological experiments is a significant improvement for myriad of cell analyses over conventional methods, which require bulk samples providing only averaged information on cell metabolism. We report on a device that integrates mobile magnetic trap array with microfluidic technology to provide, combined functionality of separation of immunomagnetically labeled cells or magnetic beads and their encapsulation with reagents into pico-liter droplets. This scheme of simultaneous reagent delivery and compartmentalization of the cells immediately after sorting, all performed seamlessly within the same chip, offers unique advantages such as the ability to capture cell traits as originated from its native environment, reduced chance of contamination, minimal use and freshness of the reagent solution that reacts only with separated objects, and tunable encapsulation characteristics independent of the input flow. In addition to the demonstrated preliminary cell viability assay, the device can potentially be integrated with other up- or downstream on-chip modules to become a powerful single-cell analysis tool.
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