# US 10,870,111 B2 · Fluidic devices with bead well geometries with spatially separated bead retention and signal detection segments and related methods > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,870,111 B2 - Document ID: US10870111B2 - Status: Granted - Kind: B2 - Application number: 15/742,616 - Filed: 2016-07-19 - Published: 2019-02-21 - Granted: 2020-12-22 - Earliest claimed: 2015-07-22 - Assignees: The University of North Carolina at Chapel Hill - Applicants: The University of North Carolina at Chapel Hill - Inventors: John Michael Ramsey; William Hampton Henley - Web page: https://patentgrep.com/patent/US10870111B2 ## CPC - B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules} - G01N 35/00029: {provided with flat sample substrates, e.g. slides (G01N35/028 takes precedence)} - B01J 2219/00317: Microwell devices, i.e. having large numbers of wells - B01J 2219/00648: by the use of solid beads - B01J 2219/00702: Processes involving means for analysing and characterising the products - B01L 2200/0668: Trapping microscopic beads - B01L 2300/0877: Flow chambers - B01L 2300/0893: having a very large number of wells, microfabricated wells - C12Q 1/6823: Release of bound markers - C12Q 1/686: Polymerase chain reaction [PCR] - G01N 2035/00158: {Elements containing microarrays, i.e. "biochip"} - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract A fluidic device includes a plurality of reaction wells, typically in a dense array, with at least one bead retention segment in fluid communication with and spatially separated from at least one signal detection segment. A respective bead retention segment can be configured to hold a single bead, which can have a reagent attached thereto. ## Claims ### Claim 1 A fluidic device comprising: a plurality of reaction wells that are in fluid isolation from each other and sealed from one another by a sealing agent, and characterized in that each reaction well has at least one bead retention segment and at least one spatially separated signal detection segment that is configured as at least one elongate channel in fluid communication with the at least one bead retention segment, wherein the at least one bead retention segment has a depth direction, wherein the at least one elongate channel extends away from the at least one bead retention segment in a length direction that is perpendicular to the depth direction, and wherein the at least one elongate channel has a length in the length direction that is greater than a width of the at least one bead retention segment; and a plurality of magnetic beads, wherein one magnetic bead of the plurality of magnetic beads is physically retained in each bead retention segment of the plurality of reaction wells. ### Claim 2 (depends on 1) The device of claim 1, wherein the at least one elongate channel has a width that is less than the width of an adjacent bead retention segment. ### Claim 3 (depends on 1) The device of claim 1, wherein the at least one bead retention segment of at least some of the plurality of reaction wells consists of a single bead retention segment. ### Claim 4 (depends on 1) The device of claim 1, wherein the at least one bead retention segment is a plurality of bead retention segments. ### Claim 5 (depends on 1) The device of claim 1, wherein at least one of the plurality of reaction wells is configured so that the at least one bead retention segment is a plurality of bead retention segments with at least one neighboring pair of bead retention segments being spaced apart by the at least one elongate channel and with the at least one elongate channel fluidly connecting a corresponding neighboring pair of bead retention segments, and wherein the at least one elongate channel has a width that is narrower or a depth that is shallower or a width that is narrower and a depth that is shallower than that of the corresponding neighboring pair of bead retention segments. ### Claim 6 (depends on 1) The device of claim 1, wherein the at least one bead retention segment is a plurality of bead retention segments having a common size. ### Claim 7 (depends on 1) The device of claim 1, wherein the at least one bead retention segment is a plurality of bead retention segments, at least one of which has a different size from at least another one. ### Claim 8 (depends on 1) The device of claim 1, wherein the at least one bead retention segment and the at least one signal detection segment of each of the reaction wells comprise tapered and/or straight walls formed into a first planar substrate of the device. ### Claim 9 (depends on 1) The device of claim 1, wherein one or more of the at least one elongate channel comprises an annular fluid channel at an end portion thereof that is spaced a distance apart from the at least one bead retention segment and in fluid communication with the at least one bead retention segment. ### Claim 10 (depends on 1) The device of claim 1, wherein at least one of the reaction wells is configured with first and second bead retention segments as the at least one bead retention segment, wherein the first and second bead retention segments are spaced apart and fluidly connected by a straight fluid channel that defines the at least one elongate channel, wherein the straight fluid channel has a narrower width than the width of the first and second bead retention segments, and wherein the first and second bead retention segments have a circular perimeter. ### Claim 11 (depends on 10) The device of claim 10, wherein the first bead retention segment is larger or smaller in diameter and/or depth compared to the second bead retention segment. ### Claim 12 (depends on 1) A method for analyzing an analyte using the device of claim 1 by electronically detecting a signal from the at least one signal detection segment of the plurality of reaction wells, wherein reagents and/or analytes released from the beads held in respective bead retention segments diffuse through a common solution volume of respective reaction wells, and wherein the signal is detected from a signal detection segment of the at least one signal detection segment while the target beads are held in the respective bead retention segments with the signal detection segment of the at least one signal detection segment generating an optical signal comprising a tail that is distinguishable from a background signal of the corresponding bead. ### Claim 13 (depends on 12) The method of claim 12, wherein the signal comprises a fluorescence signal. ### Claim 14 (depends on 12) The method of claim 12, wherein electronically detecting the signal is carried out for an array in which one or more of the plurality of reaction wells comprise a different type of bead from one or more other ones of the plurality of reaction wells, each type of bead encoded with a unique signature and each type of bead functionalized with a different reagent for allowing detection of different analytes or different chemical reactions or different analytes and different chemical reactions. ### Claim 15 A fluidic device comprising: a plurality of reaction wells characterized in that each reaction well has at least one bead retention segment and at least one spatially separated signal detection segment that is configured as at least one elongate channel in fluid communication with the at least one bead retention segment, wherein the at least one bead retention segment has a depth direction, wherein the at least one elongate channel extends away from the at least one bead retention segment in a length direction that is perpendicular to the depth direction, and wherein the at least one elongate channel has a length in the length direction that is greater than a width of the at least one bead retention segment, wherein the device is a microfluidic chip and the plurality of reaction wells are provided as an array of reaction wells, wherein the at least one bead retention segment and the at least one signal detection segment reside in a first planar substrate with the first planar substrate defining an open upper surface and sidewalls of the at least one bead retention segment and the at least one signal detection segment, and wherein the device further comprises a second planar substrate that couples to the first planar substrate and defines a closed outer surface with at least one fluid port. ### Claim 16 (depends on 15) The device of claim 15, wherein each bead retention segment of the plurality of reaction wells is sized and configured to hold only a respective single bead with a diameter in a range of about 10 nm to about 1 mm, and wherein at least one of the plurality of reaction wells has a single bead retention segment as the at least one bead retention segment that merges into a separate corresponding single elongate channel as the at least one signal detection segment, and wherein the separate corresponding single elongate channel has a width that is narrower than the width of the bead retention segment and has an end portion that resides a distance L in the length direction in a range of 1 μm to 1 mm from the single bead retention segment. ### Claim 17 (depends on 15) The device of claim 15, wherein the at least one bead retention segment is sized and configured to hold a microspherical bead with a diameter in a range of 100 nm to 1 mm, and wherein the width of the at least one bead retention segment is in a range of 101% to 195% of the diameter of a respective bead held therein. ### Claim 18 A fluidic device comprising: a plurality of reaction wells characterized in that each reaction well has at least one bead retention segment and at least one spatially separated signal detection segment that is configured as at least one elongate channel in fluid communication with the at least one bead retention segment, wherein the at least one bead retention segment has a depth direction, wherein the at least one elongate channel extends away from the at least one bead retention segment in a length direction that is perpendicular to the depth direction, and wherein the at least one elongate channel has a length in the length direction that is greater than a width of the at least one bead retention segment, wherein each of the plurality of reaction wells has a volumetric capacity in a range of 1 aL to 1 μL, and wherein the device further comprises a sealing agent that seals the plurality of reaction wells from each other. ### Claim 19 A fluidic device comprising: a first substrate comprising a plurality of reaction wells characterized in that each reaction well has at least one bead retention segment and at least one spatially separated signal detection segment that is configured as at least one elongate channel in fluid communication with the at least one bead retention segment, wherein the at least one bead retention segment has a depth direction, wherein the at least one elongate channel extends away from the at least one bead retention segment in a direction that is perpendicular to the depth direction, wherein a volume capacity of each reaction well is in a range of 1 aL to 100 μL, wherein the device further comprises a spacer and a sealing agent between the first substrate and a second substrate, wherein each reaction well of the plurality of reaction wells comprises a reaction solution that does not touch the second substrate, and wherein the plurality of reaction wells are sealed from one another by the sealing agent. ### Claim 20 A fluidic device comprising: a plurality of reaction wells characterized in that each reaction well has at least one bead retention segment and at least one spatially separated signal detection segment that is configured as at least one elongate channel in fluid communication with the at least one bead retention segment, wherein the at least one bead retention segment has a depth direction, wherein the at least one elongate channel extends away from the at least one bead retention segment in a length direction that is perpendicular to the depth direction, and wherein the at least one elongate channel has a length in the length direction that is greater than a width of the at least one bead retention segment, wherein the reaction wells are provided in a density in a range of about 6,000 wells/cm2 to about 2,000,000 wells/cm2.