# US 10,500,588 B2 · Microfluidic aliquot chip for single-cell isolation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,500,588 B2 - Document ID: US10500588B2 - Status: Granted - Kind: B2 - Application number: 15/822,071 - Filed: 2017-11-24 - Published: 2018-03-15 - Granted: 2019-12-10 - Earliest claimed: 2016-01-26 - Assignees: Lidong Qin - Applicants: Lidong Qin - Inventors: Lidong Qin; Kai Zhang - Web page: https://patentgrep.com/patent/US10500588B2 ## 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} - B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip} - C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} - B01L 3/5023: {with a sample being transported to, and subsequently stored in an absorbent for analysis} - B01L 2200/0652: Sorting or classification of particles or molecules - B01L 2200/0668: Trapping microscopic beads - B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept - B01L 2300/021: Identification, e.g. bar codes - B01L 2300/0803: Disc shape - B01L 2300/0806: Standardised forms, e.g. compact disc [CD] format - B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting - B01L 2300/0893: having a very large number of wells, microfabricated wells - B01L 2300/123: Flexible; Elastomeric - B01L 2400/0457: passive flow or gravitation - B01L 2400/0487: fluid pressure, pneumatics - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - 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 According to the invention, generally, a microfluidic aliquot (MA) chip, adapted to fit in a Petri dish, has a center well (inlet) connected by branched channels to a plurality of side wells (outlets). The chip comes in various types, including a bMA Chip T1, bMA Chip T2, bMA Chip T3, and an rMA Chip. The branched channel improvement provides for a greater distance between neighboring channels and a decreased density near the center well. An insert and a base are configured to create an MA chip. ## Claims ### Claim 1 A microfluidic aliquot (MA) chip for isolating cells, comprising: a chip having a center, a top surface, a bottom surface, an outer edge and a thickness; an inlet well disposed substantially at the center of the chip, extending into and accessible from the top surface of the chip; a plurality of outlet wells disposed in an outer portion of the chip, extending into and accessible from the top surface of the chip; a plurality of multiple segments that extend from the inlet well to the outlet wells, wherein each outlet well is disposed at the end of each of the plurality of multiple segments; wherein the multiple segments comprise branched channels; wherein the chip is configured to maintain uniform distribution of a liquid and cells from the inlet well to the outlet wells; and wherein every other outlet well is disposed farther away from the inlet well than an adjacent outlet well. ### Claim 2 (depends on 1) The MA chip of claim 1, wherein the chip comprises: a plurality of first segments that form an inner section, a plurality of last segments that form an outer section, and a plurality of segments between the first segments and the last segments that form a middle section; wherein the first segment is connected to the center inlet well in a radial pattern; wherein each segment after the first segment is divided from a prior segment in a radial pattern; wherein a first channel of a last segment is shorter than a second channel of the last segment. ### Claim 3 (depends on 2) The MA chip of claim 2, wherein a plurality of segments in the middle section form a curved portion generally in the shape of a bend, whereby one of the first segments is joined to the bend of a segment in the middle section. ### Claim 4 (depends on 1) The MA chip of claim 1, wherein the chip comprises: four first segments that extend outward from the inlet well in four cardinal directions, respectively; wherein each segment branches at a 90 degree angle in two opposite directions into another segment forming a set of branched channels; and wherein each set of branched channels on a last segment correspond to a set of outlet wells. ### Claim 5 (depends on 1) The MA chip of claim 1, wherein the chip comprises: a plurality of first segments that extend outward radially from the inlet well; wherein each segment branches at a 90 degree angle in two opposite directions into another segment forming a set of branched channels; and wherein each set of branched channels on a last segment correspond to a set of outlet wells. ### Claim 6 (depends on 1) The MA chip of claim 1, wherein the chip comprises: a sheet of material selected from a group consisting of polydimethylsiloxane (PDMS), PMMA (poly(methyl methacrylate)), PS (polystyrene), and PC (polycarbonate); wherein the sheet has a top surface and a bottom surface, that correspond to the top and bottom surfaces of the chip, respectively. ### Claim 7 (depends on 1) The MA chip of claim 1, wherein the chip comprises: a center inlet well having a diameter of 2-4 mm and a volume of 3-5 μl; a plurality of side outlet wells having a diameter of 1.5-2 mm and a volume of 1-3 μl; a plurality of branched channels having a width of 50-100 μm; and wherein the chip is configured to fit within a Petri dish of 8.5-10 cm. ### Claim 8 (depends on 1) The MA chip of claim 1, wherein the chip comprises: a first layer having a top surface and a bottom surface; a second membrane layer having a top surface and a bottom surface; wherein the bottom surface of the second membrane layer has an adhesive; and wherein the bottom surface of the second membrane layer is configured to adhere to the top surface of the first layer. ### Claim 9 (depends on 8) The MA chip of claim 8, wherein the top surface of the first layer comprises: an inlet well; an outlet well array; a main-channel array; a sub-channel array; wherein the main channels are longer and wider than the sub-channels; wherein the main channels extend outward from the inlet well to the sub-channels; and wherein the sub-channels extend from the main channels to the outlet wells. ### Claim 10 (depends on 8) The MA chip of claim 8, wherein the chip comprises: a second membrane layer having a thickness of 0.03-0.3 mm; a plurality of outlet wells having a diameter of 0.5-5 mm and a height of 1-10 mm; and wherein the chip has a length of 127.8±5 mm, a width of 85.5±3 mm, and a height of 1-10 mm. ### Claim 11 (depends on 1) The MA chip of claim 1, wherein the inlet well comprises a cap having a hole that is configured to attach onto the inlet well and receive a cell suspension. ### Claim 12 (depends on 1) The MA chip of claim 1, wherein the center inlet well is configured to receive and distribute liquid through the branched channels and into the outlet wells.