Molecular diffusive membranes or materials are important for biological applications in microfluidic systems. Hydrogels are typical materials that offer several advantages, such as free diffusion for small molecules, biocompatibility with most cells, temperature sensitivity, relatively low cost, and ease of production. With the development of microfluidic applications, hydrogels can be integrated into microfluidic systems by soft lithography, flow-solid processes or UV cure methods. Due to their special properties, hydrogels are widely used as fluid control modules, biochemical reaction modules or biological application modules in different applications. Although hydrogels have been used in microfluidic systems for more than ten years, many hydrogels' properties and integrated techniques have not been carefully elaborated. Here, we systematically review the physical properties of hydrogels, general methods for gel-microfluidics integration and applications of this field. Advanced topics and the outlook of hydrogel fabrication and applications are also discussed. We hope this review can help researchers choose suitable methods for their applications using hydrogels.

It is commonly believed that bacterial chemotaxis helps cells find food. However, not all attractants are nutrients, and not all nutrients are strong attractants. Here, by using microfluidic experiments, we studiedEscherichia colichemotaxis behavior in the presence of a strong chemoattractant (e.g., aspartate or methylaspartate) gradient and an opposing gradient of diluted tryptone broth (TB) growth medium. Our experiments showed that cells initially accumulate near the strong attractant source. However, after the peak cell density (h) reaches a critical valuehc, the cells form a “escape band” (EB) that moves toward the chemotactically weaker but metabolically richer nutrient source. By using various mutant strains and varying experimental conditions, we showed that the competition between Tap and Tar receptors is the key molecular mechanism underlying the formation of the escape band. A mathematical model combining chemotaxis signaling and cell growth was developed to explain the experiments quantitatively. The model also predicted that the width w and the peak positionxpof EB satisfy two scaling relations:w/l∼(h/hc)−1/2and1−xp/l∼(h/hc)−1/2, where l is the channel length. Both scaling relations were verified by experiments. Our study shows that the combination of nutrient consumption, population growth, and chemotaxis with multiple receptors allows cells to search for optimal growth condition in complex environments with conflicting sources.

Synthetic biologists are dedicated to designing genetic systems from the bottom up to understand how living systems work. To date, a variety of genetic circuits exhibiting bistability have been designed, greatly expanding our understanding of the biological multistability in natural systems. However, the study of more complex forms of biological multistability using synthetic methods is still limited. In this report, we describe the engineering of a genetic circuit with regulatable multistability. A novel genetic toggle switch exhibiting inducible bistability and a self-activation circuit were individually designed and characterized, after which they were assembled to create a circuit that presents tristability. In bacteria, this synthetic circuit enables cells to differentiate spontaneously into three different states of gene expression. Moreover, the multistability of the circuit can be modulated by external inputs. This work provides a synthetic biology framework for the study of biological multistability and may help to understand natural multistability phenomena.

Cells live in time varying environments with multiple desirable locations separated by unfavorable regions. To study cell navigation in spatiotemporally varying environments, we developed a microfluidic “race-track” device to create traveling attractant waves with multiple peaks and a tunable wave speed (υw). We found that while the population-averaged chemotaxis drift speed (υd) increases with υw for low υw, it decreases sharply for high υw. Our single cell measurements revealed that this reversed dependence of υd on υw is caused by a “barrier-crossing” phenomenon, where a cell hops backwards from one peak attractant location to the peak behind by crossing an unfavorable (barrier) region with low attractant concentrations. The barrier-crossing process is enabled by the cell’s random motion, which acts as temperature in thermally activated processes. Our simulation results and theoretical analysis showed that the backward barrier is lowered by υw and the backward drift speed depends exponentially on υw, leading to the observed sharp drop in υd for high υw. The barrier-crossing effect is further confirmed in double well experiments.

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The biodegradation behavior of municipal solid waste (MSW) depends on the diversity and metabolic function of bacterial communities, which are affected by environmental factors. However, the diversity of the bacterial communities and metabolic functions in MSW, as well as their influencing factors, remain unclear. In deep-aged MSW, the abovementioned deficiencies are more significant, and will effectively hamper landfill disposal. In this study, high-throughput sequencing was performed to examine the bacterial community structure and metabolic function from depths of 10 m to 40 m, of two large MSW landfills on the southeast coast of China. Thermotogota (1.6–32.0%), Firmicutes (44.2–77.1%), and Bacteroidota (4.0–34.3%) were the three dominant phyla among the 39 bacterial phyla identified in aged MSW samples. Bacterial genera associated with the degradation of many macromolecules, e.g., Defluviitoga, Hydrogenispora, and Lentimicrobium were abundantly detected in MSW samples, even in aged MSW. Redundancy analysis (RDA) showed that bacterial diversity in the landfills was most strongly correlated with electrical conductivity, age, and moisture content of the MSW. Tax4fun2 analysis predicted that there were abundant metabolism functions in aged MSW, especially functional enzymes (e.g., glycine dehydrogenase and cellulase) related to amino acids and cellulose degradation. This study increases our understanding of the bacterial diversity and functional characteristics in landfilled MSW.