63 results about "Cytometry" patented technology

The present disclosure provides systems and methods for analyzing a mixed population of cells. In particular, the present disclosure provides systems and methods for digital cytometry of a biological sample, digital analysis of a biological sample, digital purification of a biological sample, evaluation of a disease in an individual, and prediction of a clinical outcome of a disease therapy.

In one embodiment, a method of building an optimized color flow cytometry panel is disclosed using a full spectrum flow cytometer with five excitation lasers and five corresponding detection modules. In another embodiment, a graphical user interface is disclosed generated by a server computer from a fluorochrome database and displayed by a client computer to assist in the selection of a set of fluorochromes for use in an assay to analyze biological samples. The GUI can display spectra graphs to visually show how fluorochromes may overlap and can generate similarity indexes for the paired fluorochrome interference and a complexity index for overall many to many interferences generated by a selected group or set of fluorochromes.

The present disclosure provides improved and useful techniques for cross-standardization of flow cytometry instruments and, particularly, spectral flow cytometry instruments. Aspects of the disclosure include methods of calibrating a flow cytometer having a plurality of fluorescence channels. Methods of interest utilize calibration sets of bead populations, wherein each bead population of the calibration set includes a different fluorophore attached to a surface thereof and the calibration set includes a number of bead populations that is less than the number of fluorescence channels of the flow cytometer. Flow cytometers, non-transitory computer-readable storage media, and kits including, e.g., calibration sets of bead populations for carrying out the subject methods are also provided.

Provided relates to the field of cytometry, specifically to flow cytometric methods and kits for improved diagnosis, prognosis and monitoring of tumors and other lesions involving immune cell infiltration. Further provided are embodiments of the subject matter which relate to compositions and methods providing high resolution quantitative means for immunophenotyping and immune modeling, and for identification of disease prognostic and therapy predictive biomarkers.

Here, the inventors have developed a spectral flow cytometry assay to measure heme biosynthesis. By using this new assay on melanoma cell lines, they observed a direct correlation between high PPIX levels in differentiated MITFhigh cells and a protection against ferroptosis. Conversely, cell lines with low PPIX levels were associated with a dedifferentiated MITFlow phenotype enriched in stem cell markers and more sensitive to ferroptosis inducers. They also found that inhibition of PPIX biosynthesis in differentiated melanoma cells synergizes with iron overload to induce lipid peroxidation and tumor cell death. Finally, they show that decreased levels of PPIX linked to increased ferroptosis sensitivity can be acquired in vivo in melanoma tumors that relapse after anti-MAPK targeted therapies. The present invention relates to a method of determining whether a subject has or is at risk of having melanoma ferroptosis sensitivity and targeted therapy resistance comprising i) determining the level of protoporphyrin IX (PPIX) in a biological sample obtained from the subject and ii) comparing the level determined at step i) with a predetermined reference value: wherein if the level of the PPIX determined at step (i) is lower than the predetermined reference value is indicative that the said patient is having melanoma ferroptosis sensitivity and targeted therapy resistance or wherein if the level of the PPIX determined at step (i) is higher than the predetermined reference value is indicative that the said patient is having melanoma ferroptosis resistance and targeted therapy sensitivity. The present invention also relates to a method for treating resistant melanoma in a subject in need thereof comprising a step of administering said subject with a therapeutically effective amount of a ferroptosis inducer.

An analytical method of and system for monitoring cell status whereby excitation energy is focused to an excitation cytometry volume in a cell sample to fluoresce first and second fluorophores of a cell. Fluorescence signals from the first and second fluorescing cell fluorophores are directed to a detection subsystem. A first peak in fluorescence intensity for the first fluorophore is detected as is a second peak in fluorescence intensity for the second fluorophore. The fluorescence signals are processed to identify when the first and second peaks occur substantially simultaneously indicating the presence of a cell at the excitation cytometry volume instead of background fluorescence and in response, the cellular status of the cell is measured using the intensity of the levels of the first and second peaks.

Described herein are techniques for predicting whether a subject will respond to an immune checkpoint inhibitor (ICI) therapy based on RNA expression data and cytometry data obtained for the subject. In some embodiments, the techniques include: obtaining the RNA expression data, the RNA expression data having been previously obtained from a tumor sample from the subject; selecting, using the RNA expression data, an MF profile type for the tumor sample; obtaining the cytometry data, the cytometry data having been previously obtained from a blood sample from the subject; determining, using the cytometry data, a G2 score for the blood sample, wherein the G2 score is indicative a likelihood that the blood sample is of a Primed (G2) immunoprofile type of multiple immunoprofile types; and predicting, based on the selected MF profile type and the G2 score, whether the subject will respond to the ICI therapy.

The invention relates to an acute leukemia (ALL) high-throughput flow cytometry kit, and belongs to the field of leukemia detection.16 ALL cell antigens are added on the basis of an original 8-color flow cytometry, 24 antigens in the same cell can be detected at the same time, the detection precision of minimal residual focus (MRD) is greatly improved, and the detection sensitivity is improved. The ALL cells are divided into 12 developmental stages, so that immunological typing during primary diagnosis and immunophenotype variation after bone marrow transplantation and CAR-T cell treatment can be more accurately recognized, and the method has very important guiding significance on evaluation of treatment effects of patients and adjustment of treatment schemes.

A method for determining whether a subject is at risk for having a progressing or high-grade pre-invasive lesion, nodule or small mass, or having a solid malignant tumour is described the method comprising: (i) determining a ratio of activated and / or exhausted T cells:naive and / or resting T cells in a sample of blood obtained from the subject, wherein the determining comprises analysing T cells using cytometry to detect the presence or absence of a panel of biomarkers comprising Ki67 and CD39, or (ii) determining a ratio of activated and / or exhausted T cells:T cells which are not activated and / or exhausted T cells in a sample of blood obtained from the subject, wherein the determining comprises analysing T cells using cytometry to detect the presence or absence of a panel of biomarkers comprising Ki67 and CD39, (iii) determining a proportion of activated and / or exhausted T cells as a percentage of T cells in a sample of blood obtained from the subject, wherein the determining comprises analysing T cells using cytometry to detect the presence of a panel of biomarkers comprising Ki67 and CD39, and / or (iv) determining a proportion of activated and / or exhausted T cells as a percentage of T cells in a sample of blood obtained from the subject, wherein the T cells are CD4 T cells, and wherein the determining comprises analysing T cells using cytometry to detect the presence of a panel of biomarkers comprising FoxP3.

An exemplary method and system is disclosed that facilitate the integration of multiplexed single-cell impedance cytometry in a high throughput format, which can be deployed upstream from microfluidic sample preparation and / or downstream to microfluidic cell separation. In exemplary method and system may employ impedance-based quantification of cell electrophysiology on the same microfluidic chip (i.e., “on-chip”) to provide distinguishing phenotypic information on the sample, without the need for additional sample handling, preparation or dilution steps as would be needed for other flow cytometry techniques.

Systems, methods, and apparatuses for performing real-time data analytics. One apparatus includes at least one electronic processor and at least one memory storing instructions executable by the at least one electronic processor. The at least one electronic processor is configured to obtain a sequence of cytometry data output by a cytometry instrument, normalize a first data point in the sequence of cytometry data using a set of normalization parameters determined based on a queue of cases belonging to a predetermined class, update the queue of cases belonging to the predetermined class with the normalized first data point in response to determining, using a machine learning model, the normalized first data point belongs to the predetermined class, and normalize a second data point in the sequence of cytometry data using an updated set of normalization parameters determined based on the updated queue of cases belonging to the predetermined class.

An objective lens unit for a microscope is disclosed. The objective lens unit is suitable to be used when at least partially submerged in a liquid sample. The objective lens unit includes an immersion lens positioned adjacent a distal front end of the enclosure of the objective lens unit. The immersion lens is arranged and configured to receive light through a front end optical aperture. An objective lens system is arranged inside an enclosure and proximal from the immersion lens. The objective lens system is configured to collect light received by the immersion lens through the front end optical aperture. The objective lens system includes a motorless automated adjustable lens, in particular a tuneable lens, configured to amend the focal length of the objective lens system relative to the immersion lens.

Systems, methods, and apparatuses for performing real-time cytometry data analysis. One apparatus includes at least one electronic processor and at least one memory storing instructions executable by the at least one electronic processor. The at least one electronic processor is configured, through execution of the instructions, to obtain flow cytometry data generated by a cytometry instrument representing cells of multiple categories, generate a feature vector representation based on the flow cytometry data using a plurality of self-organizing maps (SOMs), wherein each SOM corresponds to a different category of multiple categories, and predict each of one or more target labels of the cells by applying each of one or more regression models to the feature vector representation.

A device for directing illumination for a sample volume in microscopy, such as in-situ cytometry. The device includes a body having a distal front end with an optical aperture provided therein, and a reflector. The reflector is arranged distal from the front end and at a non-zero distance from the optical aperture and is configured to receive light emitted through the front end and reflect at least part of the light received from the front end back into the optical aperture. The reflector is connected to the body by at least one cantilevering strut. The device may be part of an objective lens unit.

An apparatus for guiding illumination for a sample volume in microscopy, in particular in in situ cytometry. The device includes a body (611) having a distal front end (62) and an optical aperture (63) disposed in the front end, and a reflector (621). And a reflector disposed distal of the front end at a non-zero distance from the optical aperture, the reflector configured to receive light emitted through the front end and to reflect at least a portion of the light received from the front end back into the optical aperture. The reflector is connected to the body by at least one cantilevered strut. The proposed device may be part of an objective unit (6).

A pump is configured for peristaltic pumping that facilitates high throughput cytometry. The pump includes a rotary head, an occlusion bed, and an engagement mechanism configured to perform functions. The functions include applying a first force that compresses a flexible tube between the occlusion bed and the rotary head in response to a second force applied to the engagement mechanism, wherein the second force is not parallel with the first force. The functions also include discontinuing applying the first force in response to a third force applied to the engagement mechanism, wherein the third force is opposite in direction to the second force.

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. 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. The detected fluorescence radiation can be used to analyze the fluorescence content of the sample, e.g., a cell / particle.

A flow cytometry system is provided. The system comprises: a flow cell or flow system configured to accommodate the flow of a fluid including one or more objects of interest; a light source configured to illuminate the object of interest; a plurality of tilted lenses each configured to capture a set of 2D images of the object as it flows through the flow cell or flow system and together configured to translocate the 2D images into a linear array of images perpendicular to the flow of the fluid; a sensor configured to integrate each set of 2D images; wherein the system is configured to read out and process the data from the linear array of images in order to determine one or more spatial parameters of the objects of interest. A method for 3D imaging an object is also provided.

Aspects of the present disclosure include methods for processing cytometer data, such as for group-wise analysis of the cytometer data (e.g., flow cytometry data in FCS format, mass cytometry data, genomic cytometry data). Methods according to certain embodiments include generating a compound population of events that include data accessors from cytometry data, such as where the compound population of cytometer data is from two or more different samples retained as separate raw data files (e.g., are not concatenated to form a single combined data file). Systems having an input module for receiving cytometer data and processor with memory having instructions for practicing the subject methods are also described. Non-transitory computer readable storage medium is also provided.

This disclosure provides a method for identifying and counting unlabeled (native) blood cells and analytes in whole blood. In particular, this disclosure relates to devices used within the body equipped with microfluidic cytometry optical sensors and detectors that recognize analytes or histological architecture from within a patient to provide clinically actionable data related to the subject's health.

The invention provides a single-tube multi-allergen specific basophilic granulocyte detection method based on multicolor flow cytometry, and belongs to the technical field of immunodetection. Through a single-tube multicolor flow cytometry technology, rapid and synchronous quantitative analysis of various allergen-specific basophilic granulocytes in peripheral whole blood is realized, and the detection efficiency and flux are remarkably improved; the method is easy and convenient to operate, small in needed sample size and especially suitable for children and other people with blood sampling difficulty, and the limitation of clinical application is greatly reduced; by directly measuring cell populations closely related to anaphylaxis, a more intuitive and reliable auxiliary diagnosis basis is provided for IgE-mediated anaphylaxis diseases, the immune state change of a patient in the treatment process can be effectively monitored, and a new technical support is provided for clinical precise intervention and curative effect evaluation.

The invention relates to an improved purification environment and pure bacterium source extracellular vesicle method, and relates to the technical field of biology, the method comprises the following steps: S1, obtaining a first crude extraction extracellular vesicle sample, carrying out exploratory multi-layer density gradient ultracentrifugation on the first sample, and carrying out characterization analysis on each centrifuged component, to determine an enrichment zone of extracellular vesicles; and S2, obtaining an extracellular vesicle sample of a second crude extraction homologous with the extracellular vesicle sample of the first crude extraction, constructing an optimized simplified density gradient according to the enrichment regions determined in the step S1, and carrying out optimized simplified density gradient ultracentrifugation on the second sample to collect and purify the extracellular vesicles. According to the method disclosed by the invention, a standardized scheme is established for a specific sample through two steps of exploratory purification and purification after optimization, and enrichment regions of the extracellular vesicles are determined by adopting an identification method combining a transmission electron microscope and nanoflow cytometry, so that the repeatability of a purification result is improved.

Aspects of the present disclosure include methods, systems, and non-transitory computer-readable storage media for classifying cytometry image data using images of single particles, e.g., single cells. A method for classifying cytometry image data according to certain embodiments includes receiving unclassified cytometry image data including a plurality of images corresponding to an image channel, adjusting aspects of at least one of the plurality of images of the cytometry image data, and applying a model to the adjusted cytometry image data to classify the cytometry image data, the model being trained to predict the presence of particles belonging to a first category of particles in the cytometry image data. Aspects of the present disclosure also include a method for training a model to classify cytometry image data, the method including receiving flow cytometry data including unclassified cytometry image data including a plurality of images, each instance of the cytometry image data corresponding to an image channel, classifying each instance of the cytometry image data of the flow cytometry data to establish ground truth data, and adjusting aspects of at least one of the plurality of images of each instance of the cytometry image data of the ground truth data to train the model to classify the cytometry image data as data including particles belonging to a first category of particles. A system for performing the subject methods is further provided.A non-transitory computer-readable storage medium is further described.

A method for automatically adjusting gates to cytometry data of a target sample is provided. The method includes receiving training cytometry data corresponding to a plurality of training samples; generating an image of the target sample, generating images of the plurality of training samples; selecting a training sample from the plurality of training samples based on a similarity between the image of the selected training sample and the image of the target sample; calculating a transformation required to morph between the image of the selected training sample and the image of the target sample; and generating a target sample gate by applying the calculated transformation to the training gate associated with the selected training sample. A corresponding system and computer-readable medium are also provided.

The present invention relates to a method for quantifying target polynucleotides, particularly RNA molecules such as 16S rRNA, through a process that converts the abundance of target molecules into the abundance of predefined barcoded oligonucleotides The invention enables reduced nonspecific binding, protocol flexibility with pausing and re-entry points, and versatile, optionally multiplexed data acquisition including flow-cytometry-like or imaging-based measurements.

This invention discloses a zero-field-based flow cytometry fluorescence detection method and system. The method includes: when a target material specifically bound to nanodiamonds flows through the detection region of a sample channel, controlling at least one of the intensity, frequency, and on / off state of a microwave field to induce periodically varying fluorescence signals emitted by the NV centers within the nanodiamonds; the magnetic field strength in the detection region satisfies the zero-field condition; an excitation light module excites the NV centers in the detection region of the sample channel; a microwave radiation module provides a microwave field to the detection region of the sample channel, and the microwave radiation signal provided by the microwave field has a fixed frequency; a photodetector module collects the fluorescence signal at a preset collection frequency; and a data processing module determines whether the target analyte contains the biomolecule to be detected based on the fluorescence signal. The technical solution provided by this invention improves the detection sensitivity of the fluorescence detection method.

Immune profiling methods include, but are not limited to, RNA sequencing (RNAseq) and cytometry. Aspects of the present disclosure relate to methods, systems, and computer-readable storage media useful for characterizing a subject with cancer. The present disclosure is based, in part, on a method for immune profiling a subject with cancer based on analysis of leukocyte populations in the subject's peripheral blood and the subject's prognosis and / or likelihood of responding to immunotherapy.

A non-transitory processor-readable medium stores instructions that, when executed by a processor, cause the processor to receive a first array of data associated with a first cytometry instrument at a first time. The first data array includes a set of scattered signal data points including data representative of at least one low forward scattered signal output, at least one high forward scattered signal output, at least one low lateral scattered signal output, and at least one high lateral scattered signal output. At a second time subsequent to the first time, cytometry instrument parameters are adjusted based on the first data array, and / or cytometry instrument parameters are adjusted based on at least one of the first data array or a second data array associated with a second cytometry instrument different from the first cytometry instrument. A first cytometry measurement of the first cytometry instrument is coordinated with a second cytometry measurement of the second cytometry instrument.

The present invention relates to a filtration device for solid phase cytometry configured to filter a biological sample possibly containing microorganisms and to allow analysis thereof, the filtration device comprising: a filtration membrane configured to filter the biological sample, the filtration membrane having an upper surface and a lower surface, characterized in that the filtration device further comprises at least: a cover glass arranged above the upper surface of the filtration membrane, the cover glass being arranged to cover the entire upper surface of the membrane, forming an interstitial space between the cover glass and the upper surface of the filtration membrane, the interstitial space being shaped to receive the fluid.

Systems and methods for gradient profile generation and automated cell cytometry analysis are disclosed. The technology comprises a computer controller, one or more syringe pumps, an incubator comprised of one or more cell cultures, an autosampler connected to a peristaltic pump, and a multi-line switch valve in communication with the incubator. The multi-line switch valve is configured to add one or more of media, PBS, quenching reagent, or inactivation reagent to the one or more cell cultures. The computer controller mediates the injection of stimulus to the cell cultures via the one or more syringe pumps based on a computed temporal pump profile that determines the stimulus concentration over discrete time points.