Aspects of the present disclosure include methods for flow cytometrically sorting a sample with particles, such as cells, based on order of identification. Methods according to certain embodiments include introducing the sample into a flow cytometer; flowing the introduced sample in a flow stream; irradiating the sample in the flow stream with a light source; detecting light from cells in the sample flowing in the flow stream; identifying phenotypes of cells in the sample flowing in the flow stream based on one or more data signals generated from the detected light; and dynamically sorting into partitions cells of the sample that have a phenotype of a collection of predetermined phenotypes based on order of identification. Systems for practicing the subject methods are also provided. Non-transitory computer readable storage mediums are also described.

A microfluidic chip configuration wherein injection occurs in an upwards vertical direction, and fluid vessels are located below the chip in order to minimize particle settling before and at the analysis portion of the chip's channels. The input and fluid flow up through the bottom of the chip, in one aspect using a manifold, which avoids orthogonal re-orientation of fluid dynamics. The contents of the vial are located below the chip and pumped upwards and vertically directly into the first channel of the chip. A long channel extends from the bottom of the chip to near the top of the chip. Then the channel takes a short horizontal turn that nearly negates any influence of cell settling due to gravity and zero flow velocity at the walls. The fluid is pumped up to a horizontal analysis portion that is the highest channel/fluidic point in the chip and thus close to the top of the chip, which results in clearer imaging. A laser may also suspend cells or particles in this channel during analysis which prevents them from settling.

An automatic focus system for an optical microscope that facilitates faster focusing by using at least two offset focusing cameras. Each offset focusing camera can be positioned on a different side of an image forming conjugate plane so that their sharpness curves intersect at the image forming conjugate plane. Focus of a specimen can be adjusted by using sharpness values determined from images taken by the offset focusing cameras.

Aspects of the disclosure include systems for sorting particles of a sample in a flow stream (e.g., a biological sample containing cells). Systems according to certain embodiments include a flow cell configured to propagate a sample through a flow stream, a light source configured to irradiate particles of the sample in the flow stream, a photodetector configured to detect light from the irradiated particles and a support stage operationally coupled to the photodetector, where the support stage includes a closed-loop feedback position encoder that is configured to adjust position in the X-Y plane in response to a data signal generated by the photodetector in response to light from the irradiated particles. Methods for sorting particles using the subject systems are also described. Non-transitory computer readable storage medium are also provided.

Particles such as blood cells can be categorized and counted by a digital image processor. A digital microscope camera can be directed into a flowcell defining a symmetrically narrowing flowpath in which the sample stream flows in a ribbon flattened by flow and viscosity parameters between layers of sheath fluid. A contrast pattern for autofocusing is provided on the flowcell, for example at an edge of a rear illumination opening. The image processor assesses focus accuracy from pixel data contrast. A positioning motor moves the microscope and/or flowcell along the optical axis for autofocusing on the contrast pattern target. The processor then displaces microscope and flowcell by a known distance between the contrast pattern and the sample stream, thus focusing on the sample stream. Blood cell images are collected from that position until autofocus is reinitiated, periodically, by input signal, or when detecting temperature changes or focus inaccuracy in the image data.

In one aspect, a method of sorting cells in a flow cytometry system is disclosed, which includes illuminating a cell with radiation having at least two optical frequencies shifted from one another by a radiofrequency to elicit fluorescent radiation from the cell, detecting the fluorescent radiation to generate temporal fluorescence data, and processing the temporal fluorescence data to arrive at a sorting decision regarding the cell without generating an image (i.e., a pixel-by-pixel image) of the cell based on the fluorescence data. In other words, while the fluorescence data can contain image data that would allow generating a pixel-by-pixel fluorescence intensity map, the method arrives at the sorting decision without generating such a map. In some cases, the sorting decision can be made with a latency less than about 100 microseconds. In some embodiments, the above method of sorting cells can have a sub-cellular resolution, e.g., the sorting decision can be based on characteristics of a component of the cell. In some embodiments in which more than two frequency-shifted optical frequencies are employed, a single radiofrequency shift is employed to separate the optical frequencies while in other such embodiments a plurality of different radiofrequency shifts are employed.

A method for determining, in parts, the volume of a bulk material (2) fed onto a conveyor belt (1) captures a depth image (6) of the bulk material (2), in parts, in a capturing region (4) by means of a depth sensor (3). So that bulk material can be reliably classified at conveying speeds of more than 2 m/s even in the case of overlaps without structurally complicated measures, the captured two-dimensional depth image (6) is fed to a convolutional neural network trained in advance, which has at least three convolutional layers lying one behind the other and a downstream volume classifier (20), the output value (21) of which is output as the bulk material volume present in the capturing region (4).

A holographic imaging system may include an optical source configured to output a source beam, a splitter configured to split the source beam into a reference beam and an object beam that is incident on a target to form a scattered object beam, and a pre-filter comprising a telecentric lens and a spatial filter. The pre-filter may be configured to receive the scattered object beam and filter diffuse light from the scattered object beam to form a filtered scattered object beam. The system may also include a combiner configured to combine the filtered scattered object beam with the reference beam to form an interference beam, and an imaging array configured to receive the interference beam and generate raw holographic data based on the interference beam.

A cell analysis device includes a determination target identifier extraction portion configured to extract a determination target identifier that is information indicating an identification substance associated with a determination target cell that is a cell of a determination target from a table indicating a corresponding relationship between the cell and the identification substance for each compartment with respect to compartments flowing along a flow path including the cell and the identification substance that is a substance associated with the cell, an identifier acquisition portion configured to acquire the identifier that is the information for identifying the identification substance included in the compartments flowing along the flow path, a determination portion configured to determine a compartment including the determination target cell among the compartments flowing along the flow path on the basis of the identifier acquired by the identifier acquisition portion and the determination target identifier extracted by the determination target identifier extraction portion, and an output portion configured to output a determination result of the determination portion.

The present invention relates to a method for cancer cell separation, and more specifically, relates to a method for cancer cell separation using micro-vibration.