Covalently modified surfaces, kits, and methods of preparation and use
Inventors
Lowe, JR., Randall D. • MASTROIANNI, Alexander J. • White, Mark P. • Lavieu, Gregory G. • Beaumont, Kristin G.
Assignees
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Publication Number
US-11801508-B2
Publication Date
2023-10-31
Expiration Date
Abstract
In biosciences and related fields, it can be useful to modify surfaces of apparatuses, devices, and materials that contact biomaterials such as biomolecules and biological micro-objects. Described herein are surface modifying and surface functionalizing reagents, preparation thereof, and methods for modifying surfaces to provide improved or altered performance with biomaterials.
Core Innovation
The invention provides a microfluidic device including an enclosure with a base, a cover, and microfluidic circuit material defining a fluidic circuit. The fluidic circuit comprises a flow region and sequestration pens, and at least one surface of the flow region is modified with a plurality of first covalently bound surface modifications, each including a first linking group and a first moiety that is a first surface contact moiety.
At least one surface of the sequestration pens is modified with a plurality of second covalently bound surface modifications, each including a second linking group and a second moiety that is a second surface contact moiety or a second reactive moiety. The first linking group and the second linking group are different from each other and/or the first moiety is different from the second moiety, so the flow region and sequestration pens present different covalently bound surface modification chemistries.
The sequestration pens include an isolation region and a connection region with a proximal opening to the flow region, fluidically connecting the isolation region to the flow region. The invention also describes a method of forming different covalently modified surfaces in a regioselective manner within a microfluidic device, where a first modifying reagent is flowed through the flow region under conditions such that it does not enter the isolation region and is reacted with covalently bound reactive moieties on at least one surface of the flow region.
Claims Coverage
The input items identify two independent claims directed to a microfluidic device and a regioselective method. Across the claims, the coverage centers on distinct covalently bound surface modifications in the flow region versus sequestration pens, with sequestration pen architecture supporting region-selective modification.
Microfluidic device with different covalently bound surface modifications for flow region and sequestration pens
A microfluidic device comprising an enclosure with a base, a cover, and microfluidic circuit material defining a fluidic circuit having a flow region and sequestration pens, where at least one surface of the flow region has a plurality of first covalently bound surface modifications with a first linking group and a first moiety that is a first surface contact moiety, and where at least one surface of the sequestration pens has a plurality of second covalently bound surface modifications with a second linking group and a second moiety that is a second surface contact moiety or a second reactive moiety, wherein the first linking group and the second linking group are different from each other and/or the first moiety is different from the second moiety.
Sequestration pens with isolation region and connection region
The sequestration pens include an isolation region and a connection region having a proximal opening to the flow region and fluidically connecting the isolation region to the flow region.
Regioselective formation of modified surfaces without extending into an isolation region
A method of forming different covalently modified surfaces in a regioselective manner within a microfluidic device, where a first modifying reagent is flowed through the flow region under conditions such that the first modifying reagent does not enter the isolation region of the sequestration pen, and the first modifying reagent is reacted with covalently bound reactive moieties on at least one surface of the flow region, thereby forming a first modified surface within the flow region that does not extend into the isolation region.
The claim coverage centers on a microfluidic circuit with a flow region and sequestration pens, where covalently bound surface modifications are distinguished by different linking groups and/or different moieties. It further covers a regioselective method that confines formation of a modified surface to the flow region by preventing modifying reagent entry into an isolation region.
Stated Advantages
Inhibits migration of motile cells out of the sequestration pens.
Reduced water contact angle after covalent surface modification.
Increased surface layer thickness after reaction.
Cell adhesion observations in sequestration pens following downstream biological functionalization with streptavidin/biotin to attach fibronectin/laminin.
Region-confined formation of covalently modified surfaces, where the modified surface does not extend into the isolation region of the sequestration pen.
Covalently bound surface modifications can be different between the flow region and the sequestration pens by using different linking groups and/or different moieties.
Sequestration pens and flow region are configured to support regioselective surface modification by preventing modifying reagent entry into the isolation region.
Documented Applications
Functionalization of microfluidic device surfaces for downstream biological functionalization, including attachment of fibronectin/laminin via streptavidin/biotin in sequestration pens.
Cell-related behavior in sequestration pens, including HeLa cells and inhibition or control of motile-cell migration out of the sequestration pens.
Export of cultured cells without laser initiation of displacement.
Bioscience applications using a microfluidic surface-modification system that provides different covalently bound surface modifications on microfluidic device flow regions versus sequestration pens.
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