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-11666913-B2
Publication Date
2023-06-06
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
A microfluidic device includes an enclosure having a flow region and a sequestration pen comprising 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 an in situ-generated isolation structure is disposed within the connection region. The in situ-generated isolation structure comprises a solidified polymer network and is configured to block passage of a micro-object between the sequestration pen and the flow region.
The solidified polymer network has a dimension of at least 50 microns across the width of the connection region, and the connection region comprises a width of about 50 to about 100 microns at the proximal opening. The solidified polymer network can be formed from a photoinitiated polymer and can be disposed next to the proximal opening or the distal opening of the connection region. The structure is therefore positioned to control micro-object passage through the connection region between the sequestration pen and the flow region.
The solidified polymer network can be degraded or displaced by hydrolysis, proteolysis, osmotic change, temperature change, optical illumination, or increased fluidic flow. The device can further include conditioned inner surfaces and coating chemistries, including polymer-based coatings, and can include an electrode configured to generate dielectrophoresis forces within the enclosure.
Claims Coverage
The independent claim coverage centers on a microfluidic device architecture with a flow region and a sequestration pen having a connection region that contains an in situ-generated isolation structure formed from a solidified polymer network. The claims include structural limitations on the connection region and isolation structure, along with dependent features for material formation, placement, actuation, and removal behavior. The consolidated set reflects two independent claim formulations with overlapping core features.
In situ-generated isolation structure blocking passage within a sequestration pen connection region
A microfluidic device comprising an enclosure with a flow region, a sequestration pen comprising an isolation region and a connection region with a proximal opening to the flow region and a distal opening to the isolation region, and an in situ-generated isolation structure disposed within the connection region and configured to block passage of a micro-object between the sequestration pen and the flow region.
Solidified polymer network isolation structure
The in situ-generated isolation structure comprises a solidified polymer network and has a dimension of at least 50 microns across the width of the connection region.
Photoinitiated polymer formation
The solidified polymer network comprises a photoinitiated polymer.
Specified connection-region width and placement
The connection region comprises a width of about 50 to about 100 microns, including about 50 to about 60 microns at the proximal opening, and the in situ-generated isolation structure is disposed next to the proximal opening or the distal opening of the connection region.
Polymer network degradation or displacement by listed triggers
The solidified polymer network can be degraded or displaced by hydrolysis, proteolysis, osmotic change, temperature change, optical illumination, or increased fluidic flow.
Electrode configured to generate dielectrophoresis forces
The microfluidic device includes a substrate with an electrode configured to generate dielectrophoresis (DEP) forces within the enclosure.
The claims cover an in situ-generated isolation structure made from a solidified polymer network, disposed within a sequestration pen connection region to block micro-object passage, with dependent limitations on photoinitiated polymer formation, connection-region width, placement, DEP actuation, and trigger-based degradation or displacement.
Stated Advantages
Enables selective isolation between the sequestration pen and the flow region by blocking passage of a micro-object.
Isolation structures can be removed or resized by increased flow, hydrolysis, proteolysis, osmotic change, temperature change, or optical illumination.
Conditioned inner surfaces and coating chemistries are configured to maintain cell viability.
Documented Applications
Selective isolation and micro-object control between a sequestration pen and a flow region in microfluidic/nanofluidic systems.
Sorting and concentration functions.
Clonal cell culturing/assaying and cell isolation/selection workflows.
Directed laminar flows and barrier-module-based operation.
Assaying cells using the microfluidic sequestration and barrier structures.
Isolating micro-objects in sequestration pens using in situ-generated isolation structures within a microfluidic device, including isolation of cells and clonal populations.
Interested in licensing this patent?