Methods and compositions for making bacteriocins are described in some embodiments herein. In some embodiments, pro-polypeptide comprising the bacteriocins in the desired ratios in cis, and separated by cleavage sited can be produced by a microbial cell comprising a nucleic acid encoding the pro-polypeptide. In some embodiments microfluidic devices and methods for making specified mixtures of antimicrobial peptides and/or bacteriocins are described.

The present invention provides a sperm sorting chip and a method for sorting sperm using the same. Said sperm sorting chip includes: a flow channel structure sequentially configured with a gradually diverging flow field region, a main flow channel, and a gradually converging main flow channel intercommunicated with each other from a first side end to a second side end; a fluid injection port, a semen injection port, and a semen extraction port separately located at the first side end and communicated with a main input channel of the gradual diverging flow field region; and a waste fluid outlet located at the second side end and communicated with the gradually converging main flow channel. The gradually diverging flow field region further includes a plurality of sub-input channels derived from the main input channel and converged into the main flow channel, and the plurality of sub-input channels have a gradually widening channel width at the junction with the main flow channel. By contrast, the gradually converging main flow channel has a gradually narrowing channel width toward the waste fluid outlet.

The present invention provides methods for delivering a transient and/or reversible complex into a cell including passing a cell suspension through a constriction, wherein said constriction deforms the cell, thereby causing a perturbation of the cell such that the complex enters the cell.

Disclosed is a method for the cell-free expression of peptides or proteins in a liquid filled digital microfluidic device. The droplets having the components required for cell-free protein expression can be manipulated by electrokinesis in order to enhance levels of protein expression in the droplets.

Provided herein are methods for evaluating tumor cell spheroids in a three-dimensional microfluidic device by determining changes in the relative levels of live cells and dead cells in aliquots cultured under different conditions. Methods described herein allow ex vivo recapitulation of the tumor microenvironment such that the in vivo effectiveness of a test compound in treating tumor tissue may be predicted.

A system for injecting a substance into one or more cells of a cell population in a tissue sample, comprising: a robotic manipulator apparatus configured to hold and position a micropipette; an injector controller; a robotic apparatus configured to manipulate a focal plane of a microscope; and a computing device configured to, for each respective cell of the one or more cells of the tissue sample: determine a 3-dimensional location of the respective cell based on images formed by the microscope and captured by a microscope camera; control the robotic manipulator apparatus to insert the micropipette into the respective cell; and control injector controller to eject the substance out of the micropipette and into the respective cell.

The present disclosure provides a method of producing uniformly sized organoids/multicellular spheroids using a microfluidic device having an array of microwells. The method involves several successive steps. First, a microfluidic device containing parallel rows of microwells that are connected with a supplying channel is filled with a wetting agent. The wetting agent is a liquid that is immiscible in water. For example, the wetting agent may be an organic liquid such as oil. In the next step, the agent in the supplying channel and the microwells is replaced with a suspension of cells in an aqueous solution that contains a precursor for a hydrogel. Next, the aqueous phase in the supplying channel is replaced with the agent, which leads to the formation of an array of droplets of cell suspension in the hydrogel precursor solution, which were compartmentalized in the wells. The droplets are then transformed into cell-laden hydrogels. Subsequently, the agent in the supplying channel is replaced with the cell culture medium continuously flowing through the microfluidic device and the cells within the hydrogels are transformed into multicellular spheroids.

Provided is an apparatus for generating a microfluidic concentration field, the apparatus including: a substrate; a base film disposed on the substrate; a microchannel, which is formed in a space between the substrate and the base film and through which a fluid flows; a through passage, which communicates with the microchannel and is configured to pass through the base film; and a membrane, which is formed at a portion where the microchannel and the through passage communicate with each other and allows the fluid flowing along the microchannel and the through passage or a material flowing together with the fluid to selectively pass through the membrane, wherein a concentration field is formed between the fluid of the through passage and the fluid of the microchannel by the membrane.

Systems, methods, and techniques are disclosed herein for isolating rare cells and clusters of cells, such as CTCs, from large volumes of sample fluids, such as whole blood, diluted blood, e g, minimally diluted blood, and other samples such as leukapheresis and aphaeresis samples. In some implementations, a microfluidic device includes a particle enrichment module and a particle separation module for iterative multistage sorting. Each module can have an array of islands in a microfluidic channel having a sample inlet at a first end of the first microfluidic channel. The array of islands is arranged in one or more rows that extend along a longitudinal direction in the microfluidic channel. Each island in a row is spaced apart from an adjacent island in the row to form a siphoning channel. The array of islands is configured and arranged to shift portions of fluid through the siphoning channel between adjacent islands.

A cell culture apparatus includes one or more plates having a first major surface and an opposing second major surface. The first major surface comprises a structured surface defining a plurality of wells. Each well has an interior surface defining an upper aperture and a nadir, wherein the upper aperture of each well has a diametric dimension in a range from 100 micrometers to 2000 micrometers. The apparatus also includes a plurality of spacers extending from the first major surface along a length of the bottom surface. A plurality of flow channels are defined between adjacent rails.