# US 2019/0376013 A1 · Microfluidic devices, systems, and methods for investigating three-dimensional cellular structures > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0376013 A1 - Document ID: US20190376013A1 - Status: Published application - Kind: A1 - Application number: 16/438,356 - Filed: 2019-06-11 - Published: 2019-12-12 - Earliest claimed: 2018-06-11 - Assignees: The University of Chicago - Applicants: The University of Chicago - Inventors: Savas TAY; Michael JUNKIN; Brooke SCHUSTER - Web page: https://patentgrep.com/patent/US20190376013A1 ## CPC - C12M 23/12: {Well or multiwell plates (C12M25/04 takes precedence)} - C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)} - 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 3/50853: {with covers or lids (closures for test tubes B01L3/50825)} - C12M 23/22: {Transparent or translucent parts (glassware for laboratory use B01L3/00)} - C12M 29/00: {Means for introduction, extraction or recirculation of materials, e.g. pumps (pumps per se F04B)} - C12N 5/0693: {Tumour cells; Cancer cells} - G01N 33/5011: {for testing antineoplastic activity} - G01N 33/5088: {of vertebrates} - B01L 3/502707: {characterised by the manufacture of the container or its components} - B01L 2300/047: Additional chamber, reservoir - B01L 2300/0874: Three dimensional network - C12N 2513/00: 3D culture - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) ## Abstract This disclosure includes microfluidic devices, systems, and methods for investigating three-dimensional (3D) cell structures (e.g., organoids, spheroids, scaffolds, hydrogels, and the like). Some examples of the present microfluidic devices comprise: a body having a first end, a second end, to lateral edges, an upper side, a lower side, a first layer region between a medial plane of the body and the lower side, and a second layer region between the medial plane and the upper side. In some such examples, the first layer region defines one or more rows of wells, with each well open at the medial plane and having a bottom between the medial plane and the lower side. In some such examples, the second layer region can defines one or more elongated channels open at the medial plane and having a top between the medial plane and the upper side, with each of the elongated channels aligned with one of the rows of wells such that the respective elongated channel is in fluid communication with each of the wells in the respective row of wells. ## Claims ### Claim 1 A microfluidic device for investigating three-dimensional (3D) cell structures, the microfluidic device comprising: a body having a first end, a second end, two lateral edges, an upper side, a lower side, a first layer region between a medial plane of the body and the lower side, and a second layer region between the medial plane and the upper side; where the first layer region defines one or more rows of wells, each well open at the medial plane and having a bottom between the medial plane and the lower side; and where the second layer region defines one or more elongated channels open at the medial plane and having a top between the medial plane and the upper side, each elongated channel of the one or more elongated channels aligned with a respective one of the rows of wells such that the elongated channel is in fluid communication with each well in the respective one of the row of wells. ### Claim 2 (depends on 1) The microfluidic device of claim 1, where the one or more rows of wells is a plurality of rows of wells, and the one or more elongated channels is a plurality of elongated channels. ### Claim 3 (depends on 1) The microfluidic device of claim 1, where: the first layer region is defined by a first material layer having a first surface and a second surface; the second layer region is defined by a second material layer having a first surface and a second surface; and the second surface of the first material layer defines the lower side of the body; the first surface of the second material layer defines the upper side of the body; and the first surface of the first material layer contacts the second surface of the second material layer along the medial plane. ### Claim 4 (depends on 1) The microfluidic device of claim 1, where the body comprises polydimethylsiloxane (PDMS). ### Claim 5 (depends on 1) The microfluidic device of claim 1, where the first layer region is coupled to a first glass slide. ### Claim 6 (depends on 5) The microfluidic device of claim 5, where a portion of the first layer region between the first glass slide and a bottom of at least one well is transparent. ### Claim 7 (depends on 1) The microfluidic device of claim 1, further comprising: a frame; where the body is clamped to the frame with the lower side of the body closer to the frame than is the upper side of the body; and where the frame is configured to permit imaging of at least one well through the lower side of the body. ### Claim 8 (depends on 1) The microfluidic device of claim 1, where the body defines one or more inlet channels and one or more outlet channels, each inlet channel extending through an exterior surface of the body to a first end of a respective one of the one or more elongated channels, and each outlet channel extending from a second end of a respective one of the one or more elongated channels through the exterior surface of the body. ### Claim 9 (depends on 8) The microfluidic device of claim 8, where the one or more inlet channels and the one or more outlet channels extend through the upper side of the body. ### Claim 10 (depends on 1) A microfluidic system for investigating three-dimensional (3D) cell structures, the microfluidic system comprising: a microfluidic device of claim 1; and a multiplexer having a plurality of multiplexer inlets, one or more multiplexer outlets, one or more waste inlets, and a plurality of valves selectively permitting fluid communication between respective multiplexer inlets and multiplexer outlet(s), each of the multiplexer outlet(s) and the waste inlet(s) in communication with at least one of the one or more elongated channels of the body of the microfluidic device. ### Claim 11 (depends on 10) The microfluidic system of claim 10, further comprising: a controller coupled to the multiplexer and configured to send signals to selectively open or close the valves to direct flow between respective multiplexer inlets and multiplexer outlet(s). ### Claim 12 (depends on 11) The microfluidic system of claim 11, where the one or more rows of wells is a plurality of rows of wells and the one or more elongated channels is a plurality of elongated channels, and the multiplexer and the controller are configured to deliver different fluid compositions to different channels and their respective wells. ### Claim 13 A method of investigating three-dimensional (3D) cell structures, the method comprising: exposing cells to culture medium in a well of a first material layer, the first material layer having a lower side and an upper side through which the well is open; coupling a second material layer to the first material layer such that a lower side of the second material layer faces the upper side of the first material layer, an elongated channel open through the lower side of the second material layer, and the second material layer is coupled to the first material layer such that the well is substantially sealed except for being in fluid communication via the elongated channel; and directing fluid to the well of the first material layer via the elongated channel of the second material layer. ### Claim 14 (depends on 13) The method of claim 13, where: the first material layer defines a plurality of rows of wells open through the upper side, the second material layer defines a plurality of elongated channels open through the lower side of the second material layer, and the second material layer is coupled to the first material layer such that wells in each row of the plurality of rows of well are substantially sealed except for being in fluid communication with a respective one of the plurality of elongated channels; and directing fluid to the well comprises directly fluid to a plurality of wells by directing fluid to at least two channels of the plurality of elongated channels. ### Claim 15 (depends on 14) The method of claim 14, where different fluid compositions are directed to different channels and their respective wells. ### Claim 16 (depends on 13) The method of claim 13, further comprising: permitting the cells to grow in the well into a 3D cellular structure. ### Claim 17 (depends on 13) The method of claim 13, where the fluid comprises a drug. ### Claim 18 (depends on 13) The method of claim 13, where a portion of the first material layer between the lower side of the first material layer and a bottom of the well is transparent. ### Claim 19 (depends on 13) The method of claim 13, further comprising: imaging at least a portion of the cells through the lower side of the first material layer; and delivering a contrast agent to the well of the first material layer via the elongated channel of the second material layer. ### Claim 20 (depends on 13) The method of claim 13, where the cells comprise cancer cells.