Methods for performing miniaturized dynamic assays using microfluidics and related systems
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Publication Number
US-12590952-B2
Publication Date
2026-03-31
Expiration Date
Abstract
Some methods of performing an assay comprise culturing target cells in one or more channels of one or more microfluidics chips, where each of the channel(s) can have a volume that is less than or equal to 100 microliters (μL). For each of one or more test liquids that each comprise a therapeutic reagent, the test liquid can flow over the cultured target cells in at least one of the channel(s) and, while the test liquid flows over the cultured target cells, data indicative of an interaction between the target cells over which the test liquid flows and the therapeutic reagent of the test liquid can be captured.
Core Innovation
The invention relates to miniaturized dynamic flow assays using one or more microfluidic chips in which cultured target cells are disposed in one or more channels, each channel having a volume that is less than or equal to 100 microliters. The target cells comprise a first fluorescent agent having an emission spectrum with a first peak wavelength, and one or more test liquids each comprise a therapeutic reagent with a second fluorescent agent having a second peak wavelength that is at least 10% different from the first peak wavelength.
While the test liquid is flowing over the cultured target cells disposed in at least one channel, one or more sequences of images are captured using one or more cameras. Each image sequence is of an area containing at least a portion of at least one channel in which the cultured target cells are disposed and over which the test liquid flows, and capturing the sequences includes receiving light emitted by the first and second fluorescent agents at an image sensor.
The fluorescence captured from the first and second fluorescent agents is spatially separated on the camera sensor so that target-cell and therapeutic-reagent interactions can be distinguished in image sequences. The document describes examples in which lymphocytes and therapeutic cells, including CAR-T and CAR-NK, interact with target cells under flow, with image-based determination concepts including killing and/or binding based on changing fluorescence proportions in the captured sequences.
Claims Coverage
The partial content includes one independent claim. The claim covers an assay workflow that combines microfluidic culture, dynamic flow of therapeutic-reagent-containing test liquids, and sensor-level separation of emissions from two fluorescent agents having emission peak wavelengths at least 10% different.
Microfluidic culture with ≤100 µL channels and two fluorescent agents
Culturing target cells in one or more channels of one or more microfluidic chips, each channel having a volume that is less than or equal to 100 microliters, wherein the cultured target cells comprise a first fluorescent agent having a first peak wavelength, and the therapeutic reagent in each test liquid comprises a second fluorescent agent having a second peak wavelength at least 10% different than the first peak wavelength.
Flowing therapeutic-reagent test liquid over cultured target cells
For each of one or more test liquids that each comprise a therapeutic reagent with a second fluorescent agent, flowing the test liquid over the cultured target cells disposed in at least one of the channels.
Time-sequence imaging during flow within the channel area
While flowing the test liquid over the cultured target cells, capturing one or more sequences of images of an area containing at least a portion of at least one channel in which the cultured target cells are disposed and over which the test liquid flows, using one or more cameras.
Camera-sensor separation of first and second emission peaks
Capturing the image sequences comprises receiving light emitted by the first and second fluorescent agents at an image sensor, with a first portion directed to a first part of the image sensor that includes the first peak wavelength and does not include the second peak wavelength, and a second portion directed to a second part that includes the second peak wavelength and does not include the first peak wavelength.
Overall, the claim set centers on a microfluidic dynamic flow assay that distinguishes a first fluorescent agent in target cells from a second fluorescent agent in therapeutic reagents by directing respective emission peaks to different parts of a camera image sensor while capturing image sequences during flow.
Stated Advantages
The fluorescence captured from the first and second fluorescent agents is spatially separated on the camera sensor so that target-cell and therapeutic-reagent interactions can be distinguished in image sequences.
Image-based determination concepts include killing and/or binding based on changing fluorescence proportions in the captured sequences.
Documented Applications
Examples include lymphocytes and therapeutic cells, including CAR-T and CAR-NK, interacting with target cells under flow.
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