Aspects of the present disclosure include methods for determining a photodetector gain correction factor for application to flow cytometer data. Methods according to certain embodiments include detecting light with a light detection system across a horizontal axis of a flow stream, generating data signals in a photodetector channel (e.g., an imaging photodetector channel) of the light detection system at a plurality of positions across the flow stream and calculating a detector gain correction factor for each position across the flow stream in response to the generated data signals. Methods also include applying a detector gain correction factor to data signals from a photodetector channel (e.g., non-imaging photodetector channels) to generate adjusted signal intensities. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a photodetector (e.g., an imaging photodetector) for practicing the subject methods are also described. Non-transitory computer readable storage medium and integrated circuits (e.g., FPGAs) are also provided.

In a first aspect, the present invention relates to a method of producing a library of microorganisms, the method comprising the steps of: a. providing a first fluid comprising at least one single cell, b. dispersing said first fluid comprising at least one single cell in a second fluid, thereby obtaining a plurality of single-layer microfluidic droplets, wherein at least one single- layer microfluidic droplet comprises at least one single cell, wherein the second fluid is immiscible with the first fluid, c. optionally, adding to said at least one single-layer microfluidic droplet a third fluid comprising a sensing compound, wherein the third fluid is miscible with said first fluid, and wherein the third fluid is immiscible with said second fluid, d. injecting said at least one single-layer microfluidic droplet optionally comprising the sensing compound into a fourth fluid, wherein said fourth fluid is immiscible with said second fluid, thereby obtaining at least one double-layer microfluidic droplet, e. dispensing said at least one double-layer microfluidic droplet into a culture medium based on the viability of the cell, f. incubating said culture medium, thereby obtaining said library. In a second aspect, the present invention relates to a system comprising: a. a first microfluidic chip for producing a plurality of single-layer microfluidic droplets wherein at least one single-layer microfluidic droplet comprises at least one single cell, b. a first microfluidic device for collecting said plurality of single-layer microfluidic droplets, c. a second device for adding a sensing compound into said at least one single-layer microfluidic droplet comprising at least one single cell, d. a second microfluidic chip for producing a double-layer microfluidic droplet, and e. a dispensing unit. In a third aspect, the present invention relates to the use of the method according to the first aspect of the present invention in a system according to the second aspect of the present invention.

The present invention discloses a method for isolating placental trophoblast cells from cervical exfoliated cells of a pregnant woman. Based on a specific antigen or combination expressed on the surface or inside of specific trophoblast cells, the designed microfluidic sorting chip or flow cytometer is used in the method to perform cell sorting of a cell suspension of a placental trophoblast sample, thus obtaining isolated and purified placental trophoblast cells. Compared with conventional methods, the method of the present invention has the advantages of non-invasively obtaining specimens and good specificity. Moreover, the method causes low risk of infection and abortion, allows earlier sampling time and can achieve the synchronous labeling of a plurality of antigens as well as identification and sorting of characteristic fluorescence signals; and the method has greatly improved accuracy and higher reliability and broader coverage area of detection results.

It is aimed to provide a technology that enables highly accurate device performance evaluation and device adjustment in optical analysis of microparticles, using the same type of beads. The present technology provides an information processing device including an information processing unit that acquires a plurality of fluorescence intensities at a plurality of light irradiation powers for a fluorescence signal from a sample including particles labeled with a fluorescent dye having a single fluorescence intensity, recognizes an intensity range of each of the plurality of fluorescence intensities detected on the basis of a fluorescence intensity balance of the sample, and calculates information relating to sensitivity of a fluorescence detection unit.

Disclosed herein include embodiments of a system, a device, and a method for sorting a plurality cells of a sample. A plurality of raw images comprising pixels of complex values in a frequency space can be generated from a plurality of channels of fluorescence intensity data of fluorescence emissions of fluorophores, the fluorescence emissions being elicited by fluorescence imaging using radiofrequency-multiplexed excitation in a temporal space. Spectral unmixing can be performed on the raw images prior to a sorting decision being made.

Aspects of the present disclosure include reconfigurable integrated circuits for characterizing particles of a sample in a flow stream. Reconfigurable integrated circuits according to certain embodiments are programmed to calculate parameters of a particle in a flow stream from detected light; compare the calculated parameters of the particle with parameters of one or more particle classifications; classify the particle based on the comparison between the parameters of the particle classifications and the calculated parameters of the particle; and adjust one or more parameters of the particle classifications based on the calculated parameters of the particle. Methods for characterizing particles in a flow stream with the subject integrated circuits are also described. Systems and integrated circuit devices programmed for practicing the subject methods, such as on a flow cytometer, are also provided.

This relates to acoustic microfluidic systems that can generate emulsions/droplets or encapsulate particles of interest (including mammalian cells, bacteria cells, or other cells) into droplets upon detection of the particles of interest flowing in a stream of particles. The systems operate on the detect/decide/deflect principle wherein the deflection step, in a single operation, not only deflects particles of interest from a stream of particles but also encapsulates the particles of interest in an emulsion droplet. The microfluidic systems have an abrupt transition in the channel geometry from a shorter channel to a taller channel (i.e., in the shape of a ‘step’) to break the stream of the dispersed phase into a droplet upon acoustic actuation. When there is no acoustic wave present, no droplets/emulsions are generated and the stream of particles proceeds uninterrupted. The rapid actuation and post-actuation recovery employed by the microfluidic systems taught herein ensure that the vast majority of selected particles are properly deflected, that few or no empty droplets are produced, and that total throughput remains high.

Aspects of the present disclosure include methods for determining photodetector gain for a plurality of photodetectors in a light detection system. Methods according to certain embodiments include applying a reference voltage to each photodetector in the light detection system, generating a reference data signal for each photodetector at the reference voltage, irradiating with a light source the photodetectors at a plurality of different applied voltages, generating output data signals for each photodetector at each of the plurality of different voltages and calculating gain of the photodetectors at each of the plurality of different applied voltages based on the output data signals for each photodetector at each applied voltage and the reference data signal. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a plurality of photodetectors for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.

The present disclosure provides apparatuses, systems, and methods for performing particle analysis through flow cytometry at comparatively high event rates and for gathering high resolution images of particles.

A single junction sorter for a microfluidic particle sorter, the single-junction sorter comprising: an input channel, configured to receive a fluid containing particles; an output sort channel and an output waste channel, each connected to the input channel for receiving the fluid therefrom; a bubble generator, operable to selectively displace the fluid around a particle to be sorted and thereby to create a transient flow of the fluid in the input channel; and a vortex element, configured to cause a vortex in the transient flow in order to direct the particle to be sorted into the output sort channel.