In recent years, technologies capable of analyzing single cells have emerged that are transforming many fields of biological research. Herein we report how DNA-functionalized hydrogel beads can serve as a matrix to capture mRNA from lysed single cells. mRNA quantification free of pre-amplification bias is ensured by using padlock probes and rolling circle amplification followed by hybridization with fluorescent probes. The number of transcripts in individual cells is assessed by simply counting fluorescent dots inside gel beads. The method extends the potential of existing techniques and provides a general platform for capturing molecules of interest from single cells.

In recent years, technologies capable of analyzing single cells have emerged that are transforming many fields of biological research. Herein we report how DNA-functionalized hydrogel beads can serve as a matrix to capture mRNA from lysed single cells. mRNA quantification free of pre-amplification bias is ensured by using padlock probes and rolling circle amplification followed by hybridization with fluorescent probes. The number of transcripts in individual cells is assessed by simply counting fluorescent dots inside gel beads. The method extends the potential of existing techniques and provides a general platform for capturing molecules of interest from single cells.

Cell culturing, whether for tissue engineering or cell biology studies, always involves placing cells in a non-natural environment and no material currently exist that can mimic the entire complexity of natural tissues and variety of cell-matrix interactions that is found in vivo. Here, we review the vast range of hydrogels, composed of natural or synthetic polymers that provide a route to tailored microenvironments.

A wet stamping method to precisely control concentrations of enzymes and inhibitors in place and time inside layered gels is reported. By combining enzymatic reactions such as autocatalysis and inhibition with spatial delivery of components through soft lithographic techniques, a biochemical reaction network capable of recognizing the spatial distribution of an enzyme was constructed. The experimental method can be used to assess fundamental principles of spatiotemporal order formation in chemical reaction networks.

A wet stamping method to precisely control concentrations of enzymes and inhibitors in place and time inside layered gels is reported. By combining enzymatic reactions such as autocatalysis and inhibition with spatial delivery of components through soft lithographic techniques, a biochemical reaction network capable of recognizing the spatial distribution of an enzyme was constructed. The experimental method can be used to assess fundamental principles of spatiotemporal order formation in chemical reaction networks.

Here, the formation of eutectic Gallium-Indium (EGaIn) liquid-metal microdroplets, both spherical and non-spherical, in microfluidic devices at room temperature is reported. Monodisperse microdroplets were created in an aqueous polyethylene glycol (PEG) solution, in oxygenated and in deoxygenated silicone oil. The volume of the droplets depends on the channel dimensions and flow rates applied, varying between 0.5 and 4 nL. Non-spherical droplets were formed in oxygenated silicone oil due to the instantaneous formation of an oxide layer. These metal “micro-rice” droplets retained their shape and did not spontaneously reflow to form shapes of the lowest interfacial energy on egress from the channel, unlike in aqueous PEG solution and in deoxygenated silicone oil. Liquid-metal droplets with such tunable morphology can potentially be used in MEMS devices for optical and electrical switches, valves and micropumps.

Microscale, quasi-2D Au–polymer brush composite objects are fabricated by a versatile, controllable process based on microcontact printing followed by brush growth and etching of the substrate. These objects fold into 3D microstructures in response to a stimulus: crosslinked poly(glycidyl methacrylate) (PGMA) brushes fold on immersion in MeOH, and poly(methacryloxyethyl trimethylammonium chloride) (PMETAC) brushes fold on addition of salt. Microcages and microcontainers are fabricated. A multistep microcontact printing process is also used to create sheets of Au–PGMA bilayer lines linked by a PGMA film, which fold into cylindrical tubes. The bending of these objects can be predicted, and hence predefined during the synthesis process by controlling the parameters of the gold layer, and of the polymer brush.

Microdroplets in microfluidics offer a great number of opportunities in chemical and biological research. They provide a compartment in which species or reactions can be isolated, they are monodisperse and therefore suitable for quantitative studies, they offer the possibility to work with extremely small volumes, single cells, or single molecules, and are suitable for high-throughput experiments. The aim of this Review is to show the importance of these features in enabling new experiments in biology and chemistry. The recent advances in device fabrication are highlighted as are the remaining technological challenges. Examples are presented to show how compartmentalization, monodispersity, single-molecule sensitivity, and high throughput have been exploited in experiments that would have been extremely difficult outside the microfluidics platform.