The invention encompasses analyzers and analyzer systems that include a single molecule analyzer, methods of using the analyzer and analyzer systems to analyze samples, either for single molecules or for molecular complexes. The single molecule uses electromagnetic radiation that is translated through the sample to detect the presence or absence of a single molecule. The single molecule analyzer provided herein is useful for diagnostics because the analyzer detects single molecules with zero carryover between samples.

In one aspect, the present teachings provide a system for performing cytometry that can be operated in three operational modes. In one operational mode, a fluorescence image of a sample is obtained by exciting one or more fluorophore(s) present in the sample by an excitation beam formed as a superposition of a top-hat-shaped beam with a plurality of beams that are radiofrequency shifted relative to one another. In another operational mode, a sample can be illuminated successively over a time interval by a laser beam at a plurality of excitation frequencies in a scanning fashion. The fluorescence emission from the sample can be detected and analyzed, e.g., to generate a fluorescence image of the sample. In yet another operational mode, the system can be operated to illuminate a plurality of locations of a sample concurrently by a single excitation frequency, which can be generated, e.g., by shifting the central frequency of a laser beam by a radiofrequency. For example, a horizontal extent of the sample can be illuminated by a laser beam at a single excitation frequency. The detected fluorescence radiation can be used to analyze the fluorescence content of the sample, e.g., a cell/particle.

To optimize an imaging range in a depth direction in terms of a relationship with a resolution, an OCT apparatus includes a signal processor that determines a reflected light intensity distribution of an imaging object on the basis of a spectrum of a detected interference light. The signal processor performs spectrum conversion, having a conversion characteristic with which a light source spectrum is converted to a Gaussian distribution curve, on the spectrum of the interference light, and determines the reflected light intensity distribution by Fourier-transforming a spectrum resulting from the spectrum conversion. In the conversion characteristic, the light source spectrum and the Gaussian distribution curve have center wavelengths differing from each other.

A particulate matter (PM) sensor comprises a substrate forming a cavity (5), the substrate comprising a semiconductor chip (4), and a light source (1) arranged in the cavity (5). The light source (1) is adapted to emit a light beam (7). The light beam (7) forms a detection volume (8) for particulate matter (9) outside the cavity (5). Optionally, the particulate matter sensor comprises an optical element (2) delimiting the cavity (5) at one end. The optical element (2) is configured to shape the light beam (7). Further, the particulate matter sensor comprises at least one photodetector (3) that is integrated into a surface of the semiconductor chip (4). The surface into which the at least one photodetector (3) is integrated faces the detection volume (8). The at least one photodetector (3) is adapted to detect light (10) scattered by particulate matter (9) in the detection volume (8).

Provided is an observation device including: a stereo image-acquisition optical system that acquires images of cells floating in a culture fluid inside a culture vessel; and an analyzer that calculates a cell density of the cells on the basis of the images acquired by the stereo image-acquisition optical system, wherein the analyzer identifies a three-dimensional position of each of the cells included in the images and calculates the cell density on the basis of the number of cells present within a predetermined three-dimensional region.

A system for dissociating cells from a cell culture vessel. The system comprises an imaging system configured to image a plurality of cells in a cell culture vessel being dissociated from at least one surface of the cell culture vessel by at least one cell dissociation agent; and at least one controller coupled to the imaging system and configured to: control the imaging system to capture a sequence of images of at least some cells in the plurality of cells during dissociation; and identify when to neutralize the at least one cell dissociation agent using the sequence of images.

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.

The invention encompasses analyzers and analyzer systems that include a single molecule analyzer, methods of using the analyzer and analyzer systems to analyze samples, either for single molecules or for molecular complexes. The single molecule uses electromagnetic radiation that is translated through the sample to detect the presence or absence of a single molecule. The single molecule analyzer provided herein is useful for diagnostics because the analyzer detects single molecules with zero carryover between samples.

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.

Microfluidic devices, systems and techniques in connection with particle sorting in liquid, including cytometry devices and techniques and applications in chemical or biological testing and diagnostic measurements.