DEVICE FOR CELL SEPARATION AND ANALYSIS AND METHOD OF USING
First Claim
1. A microflow device for separating biomolecules, such as cells, from a sample of a bodily fluid or other liquid, which device comprises:
- a body having a flow path formed therein through which such a sample containing target biomolecules can be caused to flow, the flow path having inlet means, outlet means, and a microchannel arrangement which includes a collection region extending between said inlet and outlet means, which collection region is formed as a cavity in a flat bottom surface of said body and includes a plurality of transverse separator posts that protrude from a base surface of said cavity,a flat, rigid closure plate having a top surface that is in abutting contact with said flat bottom surface of said body and closes said flow path cavity,said posts being integral with said base surface of said collection region and extending to the top surface of said closure plate,said posts being arranged in an irregular pattern extending laterally across said flow path in said collection region so as to interrupt straight-line flow and streamlined flow of liquid through said region, anda polymeric sheet wrap encircling said body and said flat rigid plate and pressing same into surface to surface contact with each other to seal said flow path against leakage,whereby disruption of streamlined flow throughout said collection region as a result of said irregular pattern of said posts creates effective capture of target biomolecules via sequestering agents attached to said surfaces in said collection region, including said top surface of said flat plate, andwhereby removal of said wrap, following separation of target biomolecules from such a sample, allows smooth dissociation of said flat rigid closure plate with said biomolecules exposed on said top surface thereof from said body and permits ready microscopic examination and/or analysis of the separated biomolecules.
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Accused Products
Abstract
A microflow device for separating or isolating cells from a bodily fluid or other liquid sample uses a flow path where straight-line flow is interrupted by a pattern of transverse posts which are arranged across the width of a collection region in an irregular or set random pattern so as to disrupt streamlined flow. Sequestering agents, such as Abs, are attached to all surfaces in the collection region via a hydrophilic permeable hydrogel coating. The collection region is formed as a cavity in a body molded from PDMS, which flexible body is sandwiched between a glass slide or comparable flat plate and a rigid top cap plate, both of which are pressed into abutting relation with the PDMS body by a heat-shrunk polymeric sleeve. Following cell separation and washing, cells can be released from the sequestering agents and the device centrifuged to force said cells to collect adjacent the hydrogel-coated slide or plate. Slitting the polymeric sleeve allows the body to then be peeled from the slide or plate, using an integral tab, to expose the separated cells on the top surface thereof for ready microscopic examination.
94 Citations
20 Claims
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1. A microflow device for separating biomolecules, such as cells, from a sample of a bodily fluid or other liquid, which device comprises:
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a body having a flow path formed therein through which such a sample containing target biomolecules can be caused to flow, the flow path having inlet means, outlet means, and a microchannel arrangement which includes a collection region extending between said inlet and outlet means, which collection region is formed as a cavity in a flat bottom surface of said body and includes a plurality of transverse separator posts that protrude from a base surface of said cavity, a flat, rigid closure plate having a top surface that is in abutting contact with said flat bottom surface of said body and closes said flow path cavity, said posts being integral with said base surface of said collection region and extending to the top surface of said closure plate, said posts being arranged in an irregular pattern extending laterally across said flow path in said collection region so as to interrupt straight-line flow and streamlined flow of liquid through said region, and a polymeric sheet wrap encircling said body and said flat rigid plate and pressing same into surface to surface contact with each other to seal said flow path against leakage, whereby disruption of streamlined flow throughout said collection region as a result of said irregular pattern of said posts creates effective capture of target biomolecules via sequestering agents attached to said surfaces in said collection region, including said top surface of said flat plate, and whereby removal of said wrap, following separation of target biomolecules from such a sample, allows smooth dissociation of said flat rigid closure plate with said biomolecules exposed on said top surface thereof from said body and permits ready microscopic examination and/or analysis of the separated biomolecules. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of separating and examining biomolecules, such as cells, from a sample of a bodily fluid or other liquid, which method comprises the steps of:
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a. causing a sample containing target biomolecules to flow along a flow path in a body of a device, the flow path including a microchannel arrangement which includes a collection region formed as a cavity in a flat bottom surface of said body wherein a plurality of transverse separator posts protrude from a base surface of said cavity, said posts being arranged in an irregular pattern extending laterally across said flow path in said collection region so as to interrupt straight-line flow and streamlined flow of liquid through said region, said body having inlet means and outlet means connected to said microchannel arrangement, said device including a flat, rigid closure plate having a top surface that is in abutting contact with said flat bottom surface of said body and closes said flow path cavity, and said device also having a polymeric sheet wrap encircling said body and said flat rigid plate and pressing same into surface to surface contact with each other to seal said flow path against leakage, whereby disruption of streamlined flow throughout said collection region occurs as a result of said irregular pattern of said posts and creates effective capture of target biomolecules via sequestering agents attached to said surfaces in said collection region, including said top surface of said flat plate, b. removing said wrap following separation of target biomolecules from such a sample, c. smoothly dissociating said flat rigid closure plate, having said biomolecules exposed on said top surface thereof, from said body, and d. subjecting the separated biomolecules on said closure plate to microscopic examination and/or analysis. - View Dependent Claims (16, 17, 18, 19, 20)
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Specification