# US 9,006,149 B2 · High-throughput biological screening > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,006,149 B2 - Document ID: US9006149B2 - Status: Granted - Kind: B2 - Application number: 12/687,717 - Filed: 2010-01-14 - Published: 2010-12-23 - Granted: 2015-04-14 - Earliest claimed: 2009-01-15 - Assignees: The Charles Stark Draper Laboratory, Inc.; The Brigham and Women's Hospital, Inc. - Applicants: Guillermo Garcia-Cardena; Peter Mack; Jeffrey T. Borenstein; Ahmad S. Khalil; Eli J. Weinberg; Jason O. Fiering; Ernest S. Kim; William J. Adams, Jr. - Inventors: Guillermo Garcia-Cardena; Peter Mack; Jeffrey T. Borenstein; Ahmad S. Khalil; Eli J. Weinberg; Jason O. Fiering; Ernest S. Kim; William J. Adams, Jr. - Web page: https://patentgrep.com/patent/US9006149B2 ## CPC - C12M 23/12: {Well or multiwell plates (C12M25/04 takes precedence)} - C12M 35/04: {Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli} - 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} ## Abstract A high-throughput flow system includes an array of wells and a separate mechanical tip positioned within each well. Each mechanical tip is separately actuated to impart a shear stress pattern. A separate sleeve may be associated with each tip for maintaining a predetermined distance between the tip and a floor of the tip's corresponding well, with each tip being rotatable within its corresponding sleeve. Alternatively, a separate post may be associated with each tip for maintaining a predetermined distance between the tip and a floor of the tip's corresponding well, with each tip being rotatable about its corresponding post. ## Claims ### Claim 1 A high-throughput flow apparatus for use with an array of wells, the apparatus comprising: an array of mechanical tips each corresponding to one of the wells; an interface for positioning each tip within its corresponding well; a separate sleeve mechanically coupled with each tip and extending beyond a distal end thereof, each sleeve thereby being adapted to contact a floor of its corresponding well and to maintain a predetermined distance between its tip and the floor of a testing chamber, wherein the testing chamber is defined by a volume captured within an inner diameter of the sleeve, each tip rotatable within its corresponding sleeve; and a driver associated with a tip for driving the tip within its respective separate sleeve to impart a shear stress pattern in its corresponding testing chamber. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein the shear stress pattern mimics a physiological hemodynamic waveform present in the circulatory system of an organism. ### Claim 3 (depends on 2) The apparatus of claim 2, wherein the physiological hemodynamic waveform comprises at least one of an atheroprotective waveform, an atheroprone waveform, or a waveform that increases stem cell differentiation. ### Claim 4 (depends on 2) The apparatus of claim 2, wherein the organism is at least one of a human or a mouse. ### Claim 5 (depends on 1) The apparatus of claim 1, wherein the shear stress pattern comprises at least one of a steady shear stress pattern or an oscillatory shear stress pattern. ### Claim 6 (depends on 1) The apparatus of claim 1, wherein the shear stress pattern comprises temporal and spatial variations. ### Claim 7 (depends on 1) The apparatus of claim 1, wherein the shear stress pattern has a magnitude of up to 35 dynes/cm2. ### Claim 8 (depends on 1) The apparatus of claim 1, wherein each tip does not contact a surface of its corresponding well. ### Claim 9 (depends on 1) The apparatus of claim 1, wherein each of the wells comprises a feature interfacing with a complementary feature of the corresponding mechanical tip for maintaining radial alignment between the tip and an interior wall of the well. ### Claim 10 (depends on 1) The apparatus of claim 1, wherein a bottom surface of each tip has a flat center and an overall conical shape. ### Claim 11 (depends on 1) The apparatus of claim 1, wherein the driver comprises a variable-speed precision motor. ### Claim 12 (depends on 11) The apparatus of claim 11, wherein the variable-speed precision motor is a stepper motor. ### Claim 13 (depends on 1) The apparatus of claim 1 further comprising a carbon dioxide source for regulating a level of carbon dioxide in an environment surrounding the well array. ### Claim 14 (depends on 1) The apparatus of claim 1 further comprising a temperature-controlled fluid bath for the well array. ### Claim 15 (depends on 14) The apparatus of claim 14 further comprising a heater for controlling the temperature of the fluid bath. ### Claim 16 (depends on 14) The apparatus of claim 14, wherein the fluid bath is maintained at a temperature of approximately 37° C. ### Claim 17 (depends on 1) The apparatus of claim 1 wherein the wells are arranged on a standard microtiter plate or cell culture plate. ### Claim 18 (depends on 1) The apparatus of claim 1, wherein the driver is associated with at least two tips. ### Claim 19 (depends on 1) The apparatus of claim 1, wherein a separate driver is associated with each tip. ### Claim 20 A high-throughput flow apparatus for use with an array of wells, the apparatus comprising: an array of mechanical tips each corresponding to one of the wells; an interface for positioning each tip within its corresponding well; a separate post mechanically coupled with each tip and extending beyond a distal end thereof, each post thereby being adapted to contact a floor of its corresponding well and to maintain a predetermined distance between its tip and the floor of the well, each tip driven independently of its corresponding post; and a driver associated with a tip for driving the tip around its respective separate post to impart a shear stress pattern in its corresponding well. ### Claim 21 (depends on 20) The apparatus of claim 20, wherein the shear stress pattern mimics a physiological hemodynamic waveform present in the circulatory system of an organism. ### Claim 22 (depends on 21) The apparatus of claim 21, wherein the physiological hemodynamic waveform comprises at least one of an atheroprotective waveform, an atheroprone waveform, or a waveform that increases stem cell differentiation. ### Claim 23 (depends on 21) The apparatus of claim 21, wherein the organism is at least one of a human or a mouse. ### Claim 24 (depends on 20) The apparatus of claim 20, wherein the shear stress pattern comprises at least one of a steady shear stress pattern or an oscillatory shear stress pattern. ### Claim 25 (depends on 20) The apparatus of claim 20, wherein the shear stress pattern comprises temporal and spatial variations. ### Claim 26 (depends on 20) The apparatus of claim 20, wherein the shear stress pattern has a magnitude of up to 35 dynes/cm2. ### Claim 27 (depends on 20) The apparatus of claim 20, wherein each tip does not contact a surface of its corresponding well. ### Claim 28 (depends on 20) The apparatus of claim 20, wherein each of the wells comprises a feature interfacing with a complementary feature of the corresponding mechanical tip for maintaining radial alignment between the tip and an interior wall of the well. ### Claim 29 (depends on 20) The apparatus of claim 20, wherein a bottom surface of each tip has a flat center and an overall conical shape. ### Claim 30 (depends on 20) The apparatus of claim 20, wherein the driver comprises a variable-speed precision motor. ### Claim 31 (depends on 29) The apparatus of claim 29, wherein the variable-speed precision motor is a stepper motor. ### Claim 32 (depends on 20) The apparatus of claim 20 further comprising a carbon dioxide source for regulating a level of carbon dioxide in an environment surrounding the well array. ### Claim 33 (depends on 20) The apparatus of claim 20 further comprising a temperature-controlled fluid bath for the well array. ### Claim 34 (depends on 33) The apparatus of claim 33 further comprising a heater for controlling the temperature of the fluid bath. ### Claim 35 (depends on 33) The apparatus of claim 33, wherein the fluid bath is maintained at a temperature of approximately 37° C. ### Claim 36 (depends on 20) The apparatus of claim 20 wherein the wells are arranged on a standard microtiter plate or cell culture plate. ### Claim 37 (depends on 20) The apparatus of claim 20, wherein the driver is associated with at least two tips. ### Claim 38 (depends on 20) The apparatus of claim 20, wherein a separate driver is associated with each tip. ### Claim 39 A high-throughput flow apparatus for use with an array of wells, the apparatus comprising: an array of mechanical tips each corresponding to one of the wells; an interface for positioning each tip within its corresponding well; a separate sleeve associated with each tip and extending a predetermined distance beyond a distal end thereof, each sleeve thereby being adapted to contact a floor of its corresponding well and to maintain the predetermined distance between its tip and the floor of a testing chamber, wherein the testing chamber is defined by a volume captured within an inner diameter of the sleeve, each tip rotatable within its corresponding sleeve; and a driver associated a tip for driving the tip within its respective separate sleeve to impart a shear stress pattern in its testing chamber. ### Claim 40 (depends on 39) The apparatus of claim 39, wherein the separate sleeve is mechanically coupled to its respective tip. ### Claim 41 (depends on 39) The apparatus of claim 39, wherein a bottom surface of each tip has a flat center and an overall conical shape. ### Claim 42 (depends on 39) The apparatus of claim 39, wherein a separate driver is associated with each tip. ### Claim 43 A high-throughput flow apparatus for use with an array of wells, the apparatus comprising: an array of mechanical tips each corresponding to one of the wells; an interface for positioning each tip within its corresponding well; a separate post associated with each tip and extending a predetermined distance beyond a distal end thereof, each post thereby being adapted to contact a floor of its corresponding well and to maintain the predetermined distance between its tip and the floor of the well, each tip driven independently of its corresponding post; and a driver associated a tip for driving the tip around its respective separate post to impart a shear stress pattern in its corresponding well. ### Claim 44 (depends on 43) The apparatus of claim 43, wherein the separate post is mechanically coupled to its respective tip. ### Claim 45 (depends on 43) The apparatus of claim 43, wherein a bottom surface of each tip has a flat center and an overall conical shape. ### Claim 46 (depends on 43) The apparatus of claim 43, wherein a separate driver is associated with each tip.