Methods for detecting particles present in a cell composition
AssigneeJuno Therapeutics, Inc.
InventorsBrittney Jo Livingston, Hai Yue
Life of this patent
Estimating the term…
Abstract
Provided are methods of detecting, assessing or determining the presence or absence of particles, such as bead particles, present in a cell composition. Also provided are articles of manufacture and kits for use in the methods.
Claims36
A method for detecting the presence or absence of beads in a cell composition, the method comprising the steps of:
- a) incubating a sample comprising at least a portion of an input composition or a sample derived from the input composition, wherein said input composition comprises one or more cells and is likely to comprise or may comprise one or more beads in which said one or more beads is capable of emitting an autofluorescence, wherein the incubating is under conditions to induce lysis of cells to produce an output composition;
- and
- b) analyzing the output composition or a portion thereof by fluorescence imaging of the autofluorescence of the one or more beads and by bright field imaging, to obtain a fluorescent image and a brightfield image and determine the presence or absence of the one or more beads in the sample.
A method for detecting the presence or absence of beads in a cell composition, the method comprising analyzing an output composition or a portion thereof by fluorescence imaging of an autofluorescence and by brightfield imaging to determine the presence or absence of beads in the output composition, wherein the output composition is a sample of at least a portion of an input composition that has been incubated under conditions to induce lysis of cells, wherein the input composition comprises one or more of the cells and is likely to comprise or may comprise one or more beads in which said one or more beads is capable of emitting the autofluorescence.
A method for detecting the presence or absence of beads in a cell composition, the method comprising:
- (a) processing a cell composition comprising cells and beads under conditions to remove or reduce the beads of the cell composition, thereby producing an input composition comprising one or more of the cells and that may comprise one or more beads that is a residual bead, wherein the one or more beads is capable of emitting an autofluorescence;
- (b) incubating a sample comprising at least a portion of the input composition or a sample derived from the input composition, wherein the incubating is under conditions to induce lysis of the one or more cells to produce an output composition;
- (c) enriching for the one or more beads in the output composition by a step involving selecting for the one or more beads present in the output composition to produce an enriched output composition;
- and
- (d) analyzing the enriched output composition or a portion thereof by fluorescence imaging of the autofluorescence and by brightfield imaging to determine the presence or absence of residual beads in the sample.
A method for detecting the presence or absence of beads in a cell composition, the method comprising
- (i) illuminating an output composition or a portion thereof with one or more light sources to generate an illuminated sample, wherein at least one light source is capable of exciting an autofluorescence, wherein said output composition is a sample of at least a portion of an input composition that has been incubated under conditions to induce lysis of cells, wherein the input composition comprises one or more of the cells and is likely to comprise or may comprise one or more beads in which said one or more beads is capable of emitting the autofluorescence;
- (ii) capturing at least one fluorescent image, and at least one brightfield image of the illuminated sample;
- and
- (iii) analyzing the presence or absence of image objects from the one or more images for one or more parameters selected from size, area, fluorescent intensity, brightfield intensity, and/or circularity, thereby determining the presence or absence of one or more beads.
Dependencies are inferred from the claim text.