Abstract: This application describes systems and methods for assaying micro-objects in a microfluidic device. These methods include contacting a reagent with a micro-object by introducing a reagent in a first fluidic medium to a flow region of a microfluidic device, wherein the microfluidic device comprises the flow region and a chamber comprising a proximal opening fluidically connecting the chamber to the flow region, diffusing the reagent from the flow region into the chamber; introducing a micro-object into the flow region of the microfluidic device, and diffusing the reagent from the chamber to the flow region to contact the reagent with the micro-object within the flow region. Other embodiments are described.

Abstract: Methods, systems, and kits for determining a level of dissolved oxygen within a microfluidic device are provided. The microfluidic device can be suitable for cell culture. The methods, systems, and kits can further be used to determine a level of oxygen consumption in a population of biological micro-objects. In particular, the methods, systems, and kits of the present disclosure rely on flowing a fluidic medium containing a dye and a supplied partial pressure of oxygen into the microfluidic device for a period of time, wherein fluorescence emitted by the dye changes based on availability of oxygen proximate to the dye; taking a fluorescence image of an area of interest within the chamber; and correlating fluorescence of the fluorescence image of the area of interest to a reference to determine an observed partial pressure of the oxygen in the area of interest.

Abstract: Metabolic engineering has developed microbial cell factories as sustainable alternatives to chemical synthesis from petroleum feedstocks or harvesting of animals and plants, but current methods can be a costly and labor-intensive commitment. Methods are described herein for microfluidic methods of screening thousands of variant cells in order to reduce time and uncertainty to provide improved strains.

Abstract: The present disclosure relates to methods for assaying and controlling micro-objects in a microfluidic device. In situ-generated hydrogel barriers are provided for dividing a microfluidic chamber into areas where assaying a cell may be performed without interference from the presence of the cell itself.

Abstract: Cartridge for manufacturing a population of cells suitable for formulation as a cellular therapeutic are disclosed herein, along with systems for operating the cartridges and performing methods to generate the population of cells suitable for formulation as a cellular therapeutic. The population of cells suitable for formulation as a cellular therapeutic can be T-cells, including CAR T-cells. The systems and methods can be largely automated.

Abstract: Cartridges for manufacturing a population of cells suitable for formulation as a cellular therapeutic are disclosed herein, along with systems and instruments for operating the cartridges and performing methods to generate the population of cells suitable for formulation as a cellular therapeutic. The population of cells suitable for formulation as a cellular therapeutic can be immunological cells, such as T lymphocytes, including endogenous T cells (ETCs), tumor infiltrating lymphocytes (TILs), CAR T-cells, TCR engineered T-cells, or otherwise engineered T-cells. The systems and methods can be largely automated.