In situ-generated microfluidic isolation structures, kits and methods of use thereof
Inventors
Beaumont, Kristin G. • Ding, Nan-Linda • Kurz, Volker L. S. • Lionberger, Troy A. • Lowe, JR., Randall D. • Malleo, Daniele • McFarland, Andrew W. • Nevill, J. Tanner • Wang, Xiaohua
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12138626-B2
Publication Date
2024-11-12
Expiration Date
Abstract
In situ-generated microfluidic isolation structures incorporating a solidified polymer network, methods of preparation and use, compositions and kits therefor are described. The ability to introduce in real time, a variety of isolating structures including pens and barriers offers improved methods of micro-object manipulation in microfluidic devices. The in situ-generated isolation structures may be permanently or temporarily installed.
Core Innovation
The invention relates to isolating a micro-object in a microfluidic device by using an enclosure that includes a flow region and a sequestration pen with an isolation region and a connection region. The connection region has a proximal opening to the flow region and a distal opening to the isolation region, and the micro-object is disposed into the sequestration pen within this enclosure structure.
A solution comprising a flowable polymer is introduced into the enclosure, and the flowable polymer is solidified to form an in situ-generated isolation structure, formed in situ at least in the sequestration pen. The in situ-generated isolation structure is disposed within the connection region and is configured to block passage of the micro-object between the sequestration pen and the flow region, thereby isolating the micro-object.
The disclosure further covers formation of the in situ-generated isolation structure as an in situ solidified polymer network, including solidification by optical illumination to form the polymer network. Additional refinements include partial or proximal blocking behaviors and barrier structures that can be reversibly removed or reduced for release of the micro-object by modifying fluid flow and by conditions such as hydrolysis, proteolysis, osmolality change, temperature change, or optical illumination.
Claims Coverage
The independent claim coverage centers on isolating a micro-object in a microfluidic device by disposing the micro-object into a sequestration pen and then solidifying a flowable polymer to create an in situ-generated isolation structure located in a connection region that blocks micro-object passage between the sequestration pen and a flow region.
Sequestration pen geometry with proximal and distal openings
The enclosure comprises a flow region and a sequestration pen comprising an isolation region and a connection region, where the connection region has a proximal opening to the flow region and a distal opening to the isolation region.
In situ solidification of flowable polymer to form isolation structure
A solution comprising a flowable polymer is introduced into the enclosure, and the flowable polymer is solidified to form an in situ-generated isolation structure, disposed within the connection region and formed in situ at least in the sequestration pen.
Blocking passage of micro-object between sequestration pen and flow region
The in situ-generated isolation structure is configured to block passage of the micro-object between the sequestration pen and the flow region, and the micro-object is isolated using the in situ-generated isolation structure.
Optical illumination to form an in situ polymer network
The solidifying of the flowable polymer comprises optically illuminating a selected area so that polymerizing polymers of the flowable polymer forms a polymer network.
Removal or reduction of the isolation structure
The in situ-generated isolation structure is removed or reduced by modifying fluid flow and/or introducing hydrolytic and proteolytic agents and/or adjusting osmolality or temperature and/or optically illuminating the structure, followed by releasing the micro-object from the isolation structure.
The claims cover connection-region placement of an in situ-generated isolation structure that blocks micro-object passage between a sequestration pen and a flow region, achieved by disposing the micro-object into the sequestration pen and solidifying a flowable polymer within the enclosure. Dependent coverage further specifies optical illumination for forming a polymer network and removal or reduction of the isolation structure to release the micro-object.
Stated Advantages
Provides isolating of the micro-object using an in situ-generated isolation structure that blocks passage between the sequestration pen and the flow region.
Blocks passage of the micro-object between the sequestration pen and the flow region to isolate the micro-object.
Documented Applications
Isolating micro-objects within a microfluidic device for subsequent use in downstream handling within the sequestration pen, including isolation consistent with assaying, sorting, clonal evolution, and genotyping as described in the provided summary content.
Interested in licensing this patent?