High-throughput single-cell imaging, sorting, and isolation
Inventors
Bharadwaj, Rajiv • Fathollahi, Bahram
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9420148-B2
Publication Date
2016-08-16
Expiration Date
Abstract
The invention provides a method and apparatus for isolating individual target cells. The apparatus includes a body structure comprising a main channel, a collection channel, and a waste channel fluidly coupled at a first fluid junction. A plurality of trapping channels intersect the collection channel, each trapping channel having a diameter at a location adjacent to the intersection of the trapping channel with the collection channel that is less than a diameter of an individual target cell. The apparatus also includes an imaging system configured to image individual target and non-target cells within the main channel, thereby producing imaging data; a processor configured to perform real-time, multivariate analyses of the imaging data; and a directing system configured to direct the individual target cells. A pressure source is in fluid communication with one or more of the collection channel, the waste channel, the first side channel, and the second side channel.
Core Innovation
The apparatus provides a body structure including a main channel, a collection channel, and a waste channel fluidly coupled at a first fluid junction. A plurality of trapping channels intersect the collection channel, and each trapping channel has a diameter at a location adjacent to the intersection that is less than the diameter of an individual target cell. This structural arrangement supports isolating individual target cells by coupling cell trapping near the collection intersection with downstream routing via the collection and waste channels.
The apparatus includes an imaging system configured to image a plurality of individual target and non-target cells within the main channel, thereby producing imaging data. A processor performs real-time, multivariate analyses of the imaging data to identify and distinguish the cell corresponding to the imaging data as a target cell or a non-target cell. A directing system directs the individual target cells for capture in the collection channel, while a pressure source applies selected pressure conditions to enable trapping operations.
The pressure source applies a first pressure to immobilize a target cell at a trapping location of one of the plurality of trapping channels, with the first pressure selected from a positive pressure on the collection channel, a negative pressure on the waste channel, or a combination of the positive and negative pressure. A second pressure greater than the first pressure is then applied to draw one or more target cells into and through the trapping channel at whose trapping location the target cell has been immobilized. This immobilize-then-draw sequence is repeated for cells flowing through the main channel so that target cells are isolated as individual events.
Claims Coverage
The document provides two independent claims: an apparatus claim and a method claim. Across the independent claims, there are multiple inventive features combining microfluidic channel geometry for trapping, imaging with real-time multivariate classification, directing to a collection channel, and pressure-controlled immobilization followed by drawing through trapping channels using a first and second pressure relationship.
Microfluidic trapping geometry at a collection intersection
A body structure comprising a main channel, a collection channel, and a waste channel fluidly coupled at a first fluid junction; a plurality of trapping channels intersecting the collection channel, each trapping channel having a diameter at a location adjacent to the intersection that is less than a diameter of an individual target cell.
Imaging system producing imaging data for target/non-target discrimination
An imaging system configured to image a plurality of individual target and non-target cells within the main channel, thereby producing imaging data; and a processor configured to perform real-time, multivariate analyses of the imaging data to identify target cells from non-target cells.
Directing individual target cells
A directing system configured to direct the individual target cells.
Pressure-controlled immobilization and drawing with first and second pressures
A pressure source in fluid communication with one or more of the collection channel and the waste channel, operable to apply a pressure selected from a group consisting of a positive pressure on the collection channel, a negative pressure on the waste channel, and a combination of the positive pressure and the negative pressure; the pressure having a first pressure and a second pressure greater than the first pressure.
Flowing target and non-target cells with imaging and real-time multivariate analysis
Providing a body structure with a main channel, a collection channel, and a waste channel fluidly coupled at a first fluid junction, and a plurality of trapping channels intersecting the collection channel where each trapping location has a diameter less than a diameter of an individual target cell; flowing a plurality of target and non-target cells through the main channel; obtaining imaging data as the cells flow through an imaging area; using a processor to perform real-time, multivariate analysis to identify each cell as a target cell or a non-target cell.
Directed routing to the collection channel for target cells
Where the cell is identified as a target cell, directing the target cell into the collection channel.
Immobilizing target cells at trapping locations using a first selected pressure
Applying a first pressure to immobilize the target cell at the trapping location of one of the plurality of trapping channels, where the first pressure is selected from a group consisting of a positive pressure on the collection channel, a negative pressure on the waste channel, and a combination of the positive pressure and the negative pressure; and repeating until all target cells flowing through the main channel have been immobilized or until a target cell has been immobilized at the trapping location of all of the plurality of trapping channels, whichever event occurs first.
Drawing immobilized target cells through trapping channels using a second selected pressure
Applying a second pressure to draw one or more of the target cells into and through the trapping channel at whose trapping location the target cell has been immobilized, where the second pressure is selected from a group consisting of a positive pressure on the collection channel, a negative pressure on the waste channel, and the combination of the positive pressure and the negative pressure.
The independent claim coverage centers on microfluidic trapping channels with a trapping-location diameter smaller than an individual target cell, real-time multivariate imaging-based classification of target versus non-target cells, directing target cells to the collection channel, and using pressure to immobilize target cells at trapping locations and then apply a second pressure to draw immobilized target cells through the trapping channels.
Stated Advantages
Enables isolating individual target cells by trapping at trapping locations adjacent to intersections in a microfluidic body structure.
Enables real-time, multivariate analysis of imaging data to identify and distinguish target cells from non-target cells.
Enables directing individual target cells into the collection channel.
Enables immobilizing target cells at trapping locations using a first selected pressure and drawing them through trapping channels using a second pressure.
Documented Applications
High-throughput microfluidic imaging/sorting of rare target cells, including WBCs and CTCs, with directing/switching based on imaging classification.
Interested in licensing this patent?