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WO2024263765A2 - Système intégré de distribution de macromolécules dans des cellules individuelles à l'aide d'une commande et d'une automatisation de mouvement précis - Google Patents

Système intégré de distribution de macromolécules dans des cellules individuelles à l'aide d'une commande et d'une automatisation de mouvement précis Download PDF

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Publication number
WO2024263765A2
WO2024263765A2 PCT/US2024/034800 US2024034800W WO2024263765A2 WO 2024263765 A2 WO2024263765 A2 WO 2024263765A2 US 2024034800 W US2024034800 W US 2024034800W WO 2024263765 A2 WO2024263765 A2 WO 2024263765A2
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WO
WIPO (PCT)
Prior art keywords
chip
needle
cell
cell trapping
needle chip
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2024/034800
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English (en)
Other versions
WO2024263765A3 (fr
Inventor
Yu Ting Chow
Marc CHOOLJIAN
Jacek LECHOWICZ
Amir TAHMASEBIPOUR
Lewis PIETROPAOLI
Charles Nguyen
Sean HIGLEY
Kris SCHNEIDER
Lansing STOUT
Mark Trevisiol
Pat POINTER
Ben BRUBAKER
Nova SYED
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mekonos Inc
Original Assignee
Mekonos Inc
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Filing date
Publication date
Application filed by Mekonos Inc filed Critical Mekonos Inc
Publication of WO2024263765A2 publication Critical patent/WO2024263765A2/fr
Publication of WO2024263765A3 publication Critical patent/WO2024263765A3/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/32—Micromanipulators structurally combined with microscopes

Definitions

  • the present disclosure relates generally to systems, apparatuses, processes, and methods for delivering macromolecules or other materials into cells. Particular implementations leverage precise motion control and faithful integration and interfacing of system components to create a system for precise intracellular cargo delivery.
  • Cell engineering is a promising area for scientific research and medical treatment development with numerous potential applications in medicine, biology, pharmacology, plant science, and other fields.
  • a goal may be changing a cell’s functionality, internal chemistry, or genetic material by introducing new matter into the cell.
  • Mammalian cells are particularly difficult to transfect with high efficiency, viability, and recovery due to their fragility and aseptic handling requirements.
  • Current cell engineering solutions include chemical methods (e.g., lipofection, calcium phosphate precipitation), viral methods (e.g., retroviral, lentiviral, adenoviral vectors), and other physical methods (e.g., electroporation, microinjection, gene gun, or impalef ection).
  • open arrays e.g., an array containing nanoneedles and an array containing a porous membrane
  • Such alignment and/or planarization is difficult to achieve at the tolerances necessary at a nanometer scale.
  • An added difficulty is operating the arrays in a system that optimizes footprint while integrating access to liquid handling, sensors, and biosafety controls.
  • FIG. 1 illustrates a perspective view of an example system according to aspects of the present disclosure
  • FIG. 2 illustrates a perspective view of components and modules of an example system according to aspects of the present disclosure
  • FIGs. 3 A and 3B illustrate a perspective exploded view of an example needle chip handling module according to aspects of the present disclosure
  • FIG. 4 illustrates a perspective view of an example gantry stage module according to aspects of the present disclosure
  • FIG. 5 illustrates a perspective view of an example tip-tilt platform module according to aspects of the present disclosure
  • FIGs. 6A and 6B illustrate perspective views of an example microscope module according to aspects of the present disclosure
  • FIG. 7 illustrates a perspective view of an example fluidics module according to aspects of the present disclosure
  • FIG. 8 illustrates a flow diagram of an example process for performing a cell engineering workflow according to aspects of the present disclosure
  • FIGs. 9A through 9D illustrate flow diagrams of an example process for performing a cell engineering workflow according to aspects of the present disclosure
  • FIGs. 10A through 10G illustrate flow diagrams of an example process for performing a cell engineering workflow according to aspects of the present disclosure.
  • FIGs. 11A-11D illustrate a diagram of aspects of an example needle chip and cell loading process according to aspects of the present disclosure
  • FIGs. 12A-12C illustrate a diagram of an example alignment process according to aspects of the present disclosure.
  • FIG. 13 illustrates an example of a computing device that is operable to support cell engineering workflows and methods according to aspects of the present disclosure.
  • This disclosure describes systems, apparatuses, and methods for performing cell therapies and/or cell engineering with high precision. It is desirable in some cell engineering workflows to be able to insert a cargo (e.g., a chemical, mechanical, or genetic material) precisely into a single cell using microneedles.
  • a cargo e.g., a chemical, mechanical, or genetic material
  • References made in this disclosure to microneedles, microneedle structures, or microneedle chips are intended to also refer respectively to, for example, nanoneedles, nanoneedle structures, and nanoneedle chips.
  • references made in this disclosure to nanoneedles, nanoneedle structures, and nanoneedle chips are intended to also refer respectively to, for example, microneedles, microneedle structures, or microneedle chips.
  • needle chips can include, for example, microneedle chips and/or nanoneedle chips.
  • single cells may be trapped at target locations on a cell trapping chip and then microneedles on a microneedle chip may be brought precisely into contact with the cells at the target locations.
  • An example of trapping cells at a target location may include trapping cells at target locations in a porous membrane.
  • Another example of trapping cells at a target location may include trapping cells in pores in a cell trapping chip.
  • Another example may include both a porous membrane and a cell trapping chip used in combination. Other examples of trapping cells at target locations may also be possible.
  • Unaligned contact between the target locations and needles can cause damage or failure of the needles, the cell trapping chip, or both.
  • alignment in this context is very difficult.
  • the dimensions of target locations (e.g., pores on a cell trapping chip) and of the microneedles on a microneedle chip can be made in such small scales that there is little room for error in aligning the target locations with the microneedles. This can be especially true because of the high aspect ratio to many microneedle structures (e.g., microneedles can be several times taller than they are wide) — if there is even a slight a discrepancy in planar alignment between cell trapping chips and microneedle chips, the needles may miss the cell target locations.
  • the needles are fragile and cannot withstand contact with non-cellular materials (e.g., with other structures). Cell trapping chips may also be fragile. This fragility means that contact alignment is typically not a viable option for aligning the chips.
  • the microneedle chips and the cell trapping chips are optically aligned such that the chips are co-planarized, or brought into a substantially parallel plane, to within 1 pm of error in four edges.
  • Systems and modules used to handle and bring the chips into alignment may require similar precision tolerances.
  • a system may execute cell engineering workflows by bringing cells onto a cell trapping chip having a plurality of cell traps, aligning the cell trapping chip with a microneedle or nanoneedle chip having a plurality of microneedles or nanoneedles such that each cell trap of the cell trapping chip is aligned with a single microneedle on the microneedle chip, and causing the chips to come into contact such that materials can be transferred into the cells from the microneedles.
  • System 100 may include an enclosure 110 for housing components and modules of the system.
  • the enclosure 110 may include an air filter 112 (e.g., a HEPA air filtration system) configured to maintain a clean environment within the enclosure 110 suitable for working with microscopic materials and/or for facilitating a sterile environment.
  • the system 100 may include vibration damping feet 114. Vibration damping feet 114 may be made of a hard rubber (e.g., a rubber having Shore rubber hardness of 90D) to reduce the effect of external vibrations on the system 100.
  • the vibration damping feet 114 may include integrated casters (not shown) so that the system 100 may be moved if needed.
  • the enclosure 110 may also include brackets 116 so that the system 100 may be secured in place.
  • brackets 116 may prevent the system 100 from tipping over (such as, for example, accidental tip over or tip over as a result of seismic activity).
  • the enclosure 110 may include a first clean air cavity 120 configured to house elements and/or modules of the of the system 100, including a module assembly 130.
  • the individual components and modules of module assembly 130 will be described in greater detail below with respect to FIGs. 2-7.
  • the module assembly 130 of the integrated system 100 may be configured to perform at least a portion of intracellular cargo delivery workflows.
  • a work light 122 may be included in the first clean air cavity for illuminating module assembly 130.
  • the work light 122 may facilitate optical alignment processes performed by the module assembly 130 or improve user visibility during operation of the system 100.
  • the first clean air cavity 120 may be enclosed by a first door 124.
  • First door 124 may include vents 126.
  • Enclosure 110 may include a second cavity 140 for housing a fluid handling system 150.
  • the second cavity 140 may be enclosed by a second door 144.
  • Fluid handling system 150 may include a plurality of reagent containers 152, a liquid waste container 154, a pump 155, a valve 156, and a tubing network 158.
  • the plurality of reagent containers 152 may store reagents, including as non-limiting examples, payload materials such as mRNA, DNA, RNP, and proteins, alcohols such as IPA or ethanol, buffers such as phosphate- buffered saline (PBS), and/or diluted bleach.
  • payload materials such as mRNA, DNA, RNP, and proteins
  • alcohols such as IPA or ethanol
  • buffers such as phosphate- buffered saline (PBS), and/or diluted bleach.
  • PBS phosphate- buffered saline
  • At least one of the plurality of reagent containers 152 may contain cell culture media.
  • a cell culture media may be a media in which cells may be able to live and grow.
  • the media may include cells (e.g., living cells) cultured within a solution.
  • the media may be a solution containing elements useful for cell survival and growth but into which cells have not been added.
  • the media may include a solution of PBS, Dulbecco's Modified Eagle Medium, Roswell Park Memorial Institute (RPMI) medium, and/or Fetal Bovine Serum.
  • reagents stored in the plurality of reagent containers may be considered precious reagents.
  • Precious reagents may include, for example, pay load materials and/or a cell culture media.
  • reagents stored in the plurality of reagent containers may include nonprecious reagents.
  • Non-precious reagents may include, for example, alcohols, buffers, and/or sterilizing materials.
  • Liquid waste container 154 stores liquid waste from the system, including liquid waste from the module assembly 130. In various implementations, more than one liquid waste container 154 may be used. More than one liquid waste container 154 may, for example, prevent hazardous reactions between chemicals and/or reagents used during one or more processes using the system 100.
  • the fluid handling system 150 may include fluid transport devices for directing the flow of fluids within the system, such as pump 155, valve 156, and tubing network 158.
  • Pump 155 may include more than one pump.
  • Nonlimiting examples of the pump 155 may include a syringe pump, a peristaltic pump, a diaphragm pump, a vacuum pump, a pressure-based pump, and electroosmotic pump, a displacement pump and/or a piston pump.
  • a pump 155 may include, for example, a syringe integrated with a rotary valve.
  • Valve 156 may include more than one valve (e.g., rotary valves).
  • Tubing network 158 may be made of flexible tubing materials such as Teflon and/or silicone. The tubing network 158 may be in fluid communication with the module assembly 130 and the other elements of the fluid handling system 150.
  • Module assembly 200 presents an example of the module assembly 130 in FIG. 1.
  • Module assembly 200 may include a needle chip handling module 210 (e.g. a needle chip handler), a gantry module 220, a tip-tilt platform module 230 (e.g., an alignment module), a microscope module 240, a fluidics module 250, and vibration damping feet 260.
  • Needle chip handling module 210 stores needle chips (not shown) disposed in needle chip holders 212 and supports the needle chips during system processes and workflows.
  • Gantry module 220 moves needle chips, including during transport and alignment.
  • Tip-tilt platform module 230 enables the movement and angular alignment of a cell trapping chip in a cassette 252 (e.g., a cartridge that holds and/or carries a cell-trapping chip).
  • Microscope module 240 may facilitate coplanarization and optical alignment of the needle chips and cell trapping chips.
  • Fluidics module 250 may include a cell trapping chip (not shown) disposed in a cassette 252. Fluidics module also may include devices and channels for facilitating fluid transport into and out of the module assembly 200. These modules will be described in greater detail below.
  • module assembly 200 may include vibration damping feet 260.
  • Vibration damping feet 260 may mechanically isolate the module assembly 200 from at least a substantial amount of vibration.
  • vibration damping feet may dampen vibrations from the system 100 with respect to the module assembly 200 (e.g., module assembly 130).
  • Vibration damping feet 260 may be distinct from and/or in addition to vibration damping feet 114.
  • vibration damping feet 260 may be made of a hard rubber or some other suitable vibration damping material.
  • the modules of module assembly 200 to enable the precise automation of workflows contemplated by this disclosure, the modules will preferably be integrated with and/or interface with other modules in a highly precise and faithful manner.
  • the modules may be integrated together within build tolerances of 1 pm to 500 pm.
  • build tolerances for the module assembly 200 may be at least within industry standards for build tolerances. Build tolerances may also exceed the industry standard tolerance ranges.
  • the build tolerances may include machining tolerance ranges and/or registering tolerance ranges. An exemplary machining tolerance range may be between a tolerance of 150 pm and 200 pm. An exemplary range of registering tolerance may be within a tolerance of 25 pm to 200 pm.
  • Precisely integrated modules such as those discussed above may enable more precise motion control during cell engineering processes discussed more fully below with reference to FIGs. 8-10.
  • the individual modules of the module assembly 200 will now be discussed in greater detail.
  • Needle chip handling module 300 presents an example of the needle chip handling module 210 of FIG. 2.
  • Needle chip handling module 300 may include a plurality of needle chip holders 310, a hopper 320, a hopper rest 330, and a robotic gripper 340.
  • Each of the needle chip holders 310 may hold a needle chip (not shown).
  • the needle chip may include an array of microneedles or nanoneedles. Because microneedles and nanoneedles can be fragile, handling needle chips directly can cause the needles to break.
  • the needle chip holders 310 enable the chips to be handled and manipulated within the system with a significantly lowered chance of breaking the needles. Indirect handling of the needle chips may also help to preserve sterility of the needle chip.
  • the needle chip holders 310 are examples the needle chip holders 212 of FIG. 2. As illustrated In FIG. 3B, for example, the needle chip holders 310 may include a grip section 312, a stop section 314, and a needle chip holding section 316.
  • Grip section 312 enables a robotic gripper 340 to grip the needle chip holder 310.
  • Stop section 314 may prevent the robotic gripper 340 from gripping portions of the chip holder 310 where the needle chip is stored. Stop section 314 may also protrude such that the needle chip holder 310 may rest by the protruding portion of stop section 314 in a well 322 of the hopper 320.
  • the needle chip holding section 316 may grip, be adhered to, or otherwise hold the needle chip for use in the system. Needle chip holding section 316 may be at least partially hollow and may be open on an end to expose the needle chip. In other implementations, needle chip holding section 316 may be formed as a unitary structure with the needle chip disposed on an end of the needle chip holding section 316.
  • the hopper 320 may hold the plurality of needle chip holders 310 in wells 322.
  • a hopper may be any suitable container for housing needle chip holders 310.
  • the hopper 320 includes three wells 322, but a person of ordinary skill would recognize that configurations with either more or fewer than three wells could be made from this disclosure. Having a plurality of wells 322 may enable greater throughput and efficiency for the system 100.
  • the hopper 320 may be disposed in a hopper rest 330.
  • the hopper rest 330 may include a plurality of sensors 332.
  • the sensors 332 may be able to detect the presence and/or the position of a needle chip in a corresponding needle chip holder 310.
  • the plurality of sensors 332 may provide a well-defined position of the needle chip holders to the system 100, and/or to controllers for the system 100. For example, sensing the position of needle chip holders with precision may help automate cell engineering workflows.
  • Needle chip handler 300 may include a robotic gripper 340 for securely gripping the needle chip holder 310, as shown in FIG. 3B.
  • the robotic gripper 340 may include, for example, gripper forceps 342, a limit sensor 344, pneumatic connections 346, and a mounting bracket 348.
  • the gripper forceps 342 grip a needle chip holder 310 and hold it securely during system processes and workflows.
  • Limit sensor 344 may detect the force applied by the gripper forceps to the needle chip holder, and may provide feedback to a system controller (not shown) to limit or increase the amount of force applied by the gripper forceps.
  • limit sensor 344 may measure pneumatic pressure and/or some other indicator of the force applied by the gripper forceps 342.
  • the gripper forceps 342 may be activated by pneumatic pressure to achieve precision-controlled motion.
  • the robotic gripper may include pneumatic connections 346.
  • the robotic gripper may be mounted onto a vertical linear stage 416 of gantry module 400 by a mounting bracket 348, as illustrated in FIG. 4. While robotic gripper 340 is illustrated herein in FIG. 3B as a gripper that mechanically grasps needle chip holder (e.g., through pneumatic activation), robotic gripper 340 may be configured in other ways that would be known to those of ordinary skill in the art. For example, robotic gripper 340 may configured to lock into place with the chip holders 310 based on the shape of the robotic gripper, the chip holder, or a combination thereof.
  • Gantry module 400 can represent an example of the gantry module 220 of FIG. 2.
  • the gantry module 400 may be used to move robotic gripper 340 to perform cell engineering workflows.
  • gantry module 400 may enable highly precise motion of the robotic gripper to pick up, move, and/or adjust the position of needle chips.
  • the precision motion of the gantry module 400 may bring needle chips into alignment for cell engineering workflows.
  • Gantry module 400 may include linear stages, including a first horizontal linear stage 410, a second horizontal linear stage 412, a third horizontal linear stage 414, and a vertical linear stage 416.
  • Gantry module 400 may be a 4-degree of freedom (4DOF) gantry module.
  • the gantry module 400 may be configured to move gantry positions in each of the X, Y, and Z dimensions indicated by the reference coordinate system shown in FIG. 4.
  • the X dimension roughly corresponds to a direction parallel to first horizontal linear stage 410 (or roughly left-right in the view of FIG. 4)
  • the Y dimension roughly corresponds to a direction parallel to the second horizontal linear stage 412 and the third horizontal linear stage 414
  • the Z dimension roughly corresponds to a direction parallel to the vertical linear stage 416.
  • the first horizontal linear stage 410 may also be termed an X linear stage 410
  • the second horizontal linear stage 412 and the third horizontal linear stage 414 may be termed a first Y linear stage 412 and a second Y linear stage 416, respectively.
  • the linear stages 410, 412, 414, and 416 may include linear actuators 420, 422, 424, and 426 (e.g., ball screw linear actuators) to enable precise motion control of the linear stages.
  • Gantry module 400 may also rotate in a substantially horizontal plane about an axis substantially parallel to the Z dimension (e.g., yaw rotation).
  • the rotational motion of the gantry module 400 may be facilitated by rotary compliance stages 432 and 434 coupled between the X linear stage and the respective Y linear stages.
  • Rotational motion of the gantry module 400 in the horizontal plane may be achieved, for example, by extending one of the Y linear stages a different distance or at a different rate from the other Y linear stage.
  • the gantry module 400 may include sensors. Such sensors may be coupled to the linear stages and may precisely locate the position of the linear stages and/or determine the coordinates of the gantry module 400 within a coordinate frame of reference.
  • the gantry module 400 may include a first sensor 440, a second sensor 442, a third sensor 444, and a fourth sensor 446.
  • the sensors 440, 442, 444, and 446 may include, as non-limiting examples, encoders, optical sensors, mechanical sensors, and/or electromagnetic sensors.
  • the motions of the linear stages 410, 412, 414, and 416 may be controlled by a controller or plurality of controllers (not shown). In various implementations the controller or controllers may control the gantry module 400’ s motions based on feedback or other information obtained from the sensors 440, 442, 444, and 446.
  • FIG. 5 illustrates a perspective view of an example of a tip-tilt platform module 500 (e.g. an alignment module) according to aspects of the present disclosure.
  • the tip-tilt platform module 500 presents an example of the tip-tilt platform module 230 of FIG. 2.
  • the tip-tilt platform module 500 may include a platform 510.
  • the platform 510 may be a custom platform (e.g., it may be custom machined, milled, molded, or otherwise shaped).
  • Tip-tilt platform module 500 may be configured to adjust the pitch angle and/or roll angle of the platform 510 relative to the horizontal. This may allow the alignment module (e.g tip-tilt platform module) to adjust the planar angle of a chip (e.g., a cell trapping chip, or a needle chip) to align with the planar angle another chip.
  • the pitch and/or roll angle of the alignment module may be adjusted with a high degree of precision to enable planar alignment of microneedle structures.
  • the platform 510 may be supported by a first positional linear stage 520 (e.g., a first pusher) coupled to the platform 510 via a first ball bearing 522, a second positional linear stage 530 (e.g., a second pusher) coupled to the platform 510 via a second ball bearing 532, and a third positional linear stage 540 (e.g., a third pusher) coupled to the platform 510 via a third ball bearing 542.
  • Each of the pushers, including first positional linear stage 520, second positional linear stage 530 and third positional linear stage 540 may include a corresponding motor that extends or retracts the pusher. Examples of the motors are shown in FIG. 5 as first motor 524, second motor 534, and third motor 544.
  • the motor positions may range between -12.5 and 12.5 mm relative to a resting position. In various implementations, when each of the motors 524, 534, and 544 are set to the same value, the pushers points of contact at the ball bearings 522, 532, and 542 will be the same height across the platform 510 and the platform will be level.
  • the positional linear stages may form a triangle with the ball bearings located at the vertices.
  • the triangle may be an equilateral (or substantially equilateral) triangle.
  • the base length of such an equilateral triangle may range between 285 mm and 365 mm.
  • the base of the equilateral triangle may be 337.75 mm and the height of the equilateral triangle may be 292.5 mm.
  • the platform 510 may be tilted up to 5 degrees above or below the horizontal by extending and/or retracting at least one of the positional linear stages.
  • the platform 510 may include a first aperture 512, a cassette socket 514, a second aperture 516, a securing member 518, and a fastener 519.
  • First aperture 512 may accommodate a needle chip holder 310.
  • First aperture 512 may allow a microscope module (e.g., microscope module 240 of FIG. 2 or microscope module 600 of FIG. 6A) to determine the relative position of a needle chip (not shown) in the needle chip holder 310.
  • the first aperture 512 may have a diameter larger than the diameter of the needle chip holder 310 to provide sufficient clearance between the needle chip holder 310 and the walls of the first aperture 512. Such clearance may enable alignment adjustments to be made to the needle chip holder 310.
  • Cassette socket 514 may be formed in platform 510.
  • cassette socket 514 may be formed in a face of the platform 510 opposite from the positional linear stages (pushers) 520, 530, and 540.
  • Cassette socket 514 may be configured to accept a cassette (not shown), such as the cassettes described in greater detail with respect to FIG. 7.
  • the cassette may be secured in place in the cassette socket 514 via securing member 518.
  • securing member 518 may be able to be fastened in place with fastener 519 (e.g., a hand-driven screw, a bolt, a spring fastener, a tabular fastener, a latch, a locking mechanism, or the like).
  • the second aperture 516 may allow for a microscope module (such as microscope module 240 or 600) to determine the relative position of a cell trapping chip disposed in a cassette on the platform 510.
  • determining the relative position of a cell trapping chip may allow the pitch and/or roll angle of the cell trapping chip to be aligned with the pitch and roll angle of the needle chip.
  • the needle trapping chip may be formed with the same pitch of features and the same alignment fiducials as the needle chip to facilitate this alignment.
  • the tip-tilt platform module 500 can create an alignment system with six degrees of freedom for aligning the nanoneedle chip and the cell trapping chip.
  • the gantry stage module 400 can control the motion of the needle chip in the X, Y, and Z directions, and its rotational (yaw) angle.
  • a cell trapping chip can be mounted on the tip-tilt platform of the tip-tilt platform module 500, and the pushers (e.g., positional linear stages) 520, 530, and 540 can be engaged to adjust the platform 510 so that the pitch and roll of the cell-trapping chip matches the pitch and roll of the needle chip.
  • Positional linear stages 520, 530, and 540 may also be configured to adjust the Z position of the cell trapping chip.
  • Each of the pushers 520, 530, and 540 may include a respective interface connection 526, 536, and 546 such that the respective motors 524, 534, and 544 may be powered and/or controlled by a controller (not shown).
  • the interface connections 526, 536, and 546 may communicate pusher position information as feedback to the controller.
  • the controller may be configured using a motor to world (MTOW) transformation.
  • the MTOW transformation may translate a desired pitch and/or roll angle for the platform 510 into the required linear motion of the pushers 520, 530, and 540.
  • Microscope module 600 presents an example of the microscope module 240 of FIG. 2.
  • Microscope module 600 may include a plurality of linear stages (collectively referred to as an XYZ linear stage).
  • Microscope module 600 may include a first horizontal linear stage 610 (e.g., an actuator in the X dimension) driven by a first motor 612, a second horizontal linear stage 620 (e.g., an actuator in the Y dimension) driven by a second motor 622, and a linear stage actuator 630 ((e.g., an actuator in the Z.
  • microscope module 600 may also include one or more sources of illumination.
  • the microscope module will be preferably integrated precisely and faithfully into the module assembly 200 along with the other modules discussed above.
  • the two microscopes may be mounted on the linear stage actuator 630.
  • the motors 612, 622, and 632 may be configured to move the microscopes 640 and 650 along the axes of the XYZ linear stage into a position to view and/or adjust focus on the needle chips and cell trapping chips (e.g., during alignment and/or during cell engineering workflows) and to adjust the focus plane of each of the two microscopes.
  • FIG. 6B A perspective view of aspects of the microscope module 600 including low magnification microscope 640 and high magnification microscope 650 is shown at FIG. 6B.
  • the low magnification microscope 640 may be optically coupled to a first camera 642.
  • the high magnification microscope 650 may be optically coupled to a second camera 652.
  • the high magnification microscope 650 may be optically coupled to the second camera 652 via a mirror assembly 654.
  • Mirror assembly 654 may enable the second camera 652 to detect images through the high magnification microscope 650 without needing to be in the straight-line optical path of light entering the high magnification microscope 650 through its lens 656.
  • mirror assemblies may be likewise applied to the low magnification microscope 640, or may be included inside the low magnification microscope 640, such that the path light takes from lens 646 may be directed to the first camera 642 coupled to the low magnification microscope 640.
  • Configuring microscopes to use mirror assemblies like mirror assembly 654 may allow for a lower profile microscope module 600 to be used in the system 100.
  • the low magnification microscope 640 may be configured to perform rough alignment of the needle chip and the cell trapping chip. In various implementations, the low magnification microscope may be configured to evaluate cell trapping efficiency after a cell trapping process is performed.
  • the high magnification microscope 650 may be configured to accurately determine and focus on the plane of the needle chip and cell trapping chip. Accurately determining the plane of the chips may enable precise alignment of the nanoneedles on the needle chip to the previously discussed target locations (e.g., pores) of the trapped cells on the celltrapping chip. Precise planar alignment thus enables the system 100 to perform cell membrane penetration and delivery of biomaterials into cells.
  • the microscope module 600 may be controlled with an off-the-shelf control circuit (not shown) and programmed to focus on the respective fiduciary marks of the nanoneedle chip and cell trapping chip using a focus algorithm, such as the predictive focus algorithm discussed in greater detail below.
  • a focus algorithm such as the predictive focus algorithm discussed in greater detail below.
  • Fluidics module 700 presents an example of the fluidics module 250 of FIG. 2.
  • Fluidics module 700 is shown as integrated with other modules and elements of the module assembly 200, including needle chip handling module 300 and tip-tilt platform module 500 (e.g., alignment module).
  • Fluidics module 700 may include a cassette 710, an opening 716, a cell trapping chip 718, a first syringe 720, a second syringe 722, a first syringe actuator 724, a second syringe actuator 726, a fluid handling manifold 730, fluid connections 732, and a flow rate sensor 734.
  • Cassette 710 may be disposed in a cassette socket 514 formed in the surface of the platform 510.
  • Cassette 710 may also be referred to as a cartridge (e.g., a fluidic cartridge), a housing, or as a carrier for a cell trapping chip 718.
  • the cassette 710 may include a cavity or chamber for fluid transport, storage, separation, etc.
  • the cassette 710 may be secured in place in the cassette socket 514 via securing member 518. In various implementations, securing member 518 may be able to be fastened in place with fastener 519.
  • the cassette 710 may contain an opening 716. Opening 716 may expose cell trapping chip 718. Opening 716 may facilitate chip alignment or other cell engineering processes, as a microscope could view the cell trapping chip 718 through the opening.
  • the cell trapping chip 718 may include a porous membrane.
  • the porous membrane can include target positions for cells, which may include pores. Those pores, for example, can be smaller than a dimension of the cells to be trapped on the cell trapping chip. These pores can be dimensioned such that when cells are present on the cell trapping chip 718 and an air pressure difference is applied to the pores, single cells become trapped at the pores.
  • microneedles on the needle chip align with the previously discussed target locations and can impact or poke the cells to deliver biomaterials into the cells.
  • cell trapping chip 718 may be transparent or translucent.
  • a transparent chip may enable better alignment between a needle chip and the cell trapping chip, as alignment marks on the needle chip could be seen through the cell trapping chip and aligned with alignment marks on the cell trapping chip.
  • Transparent chips may also enable better determinations of cell trapping effectiveness. For example, if cells were to be trapped at target locations on the cell trapping chip, after the cell trapping, a microscope could be used to see through the cell trapping chip and determine the percentage of target locations that effectively trapped cells at their respective locations.
  • the fluidics module 700 may include syringes, such as first syringe 720 and second syringe 722, configured to deliver and/or retrieve fluids into the cassette 710 for cell engineering workflows.
  • the syringes 720 and/or 722 may deliver solutions containing cells (e.g., a blood sample, a plasma sample, or a cell culture media) onto the cell trapping chip 718.
  • pumps having a fluid connection or connections with the cassette 710 may be used in place of syringes to transport material onto or off of the cell trapping chip 718.
  • displacement pumps and/or pressure control systems may be used to transport fluids into the cassette 710.
  • Other nonlimiting examples of fluidic pumps that could be employed in fluid transport in place of and/or in addition to syringes 720/722 may include a syringe pump, a peristaltic pump, a diaphragm pump, a vacuum pump, a pressure-based pump, and electroosmotic pump, a displacement pump and/or a piston pump.
  • Other devices typical to fluid handling systems may be included, such as flow sensors, leak monitors, valves, flow regulators, and the like.
  • First syringe 720 and/or second syringe 722 may retrieve cells during a cell engineering workflow or after a cell engineering workflow has completed. Additionally or alternatively, the first syringe 720 and/or second syringe 722 may deliver and/or retrieve other fluids (e.g., buffers, reagents, cell culture media, and so on) to/from the cassette 710 for use in cell engineering workflows.
  • First syringe 720 may be actuated by a first syringe actuator 724, and second syringe 722 may be actuated by a second syringe actuator 726.
  • Fluids may also be delivered to and/or retrieved from the cassette 710 via a fluid handling manifold 730.
  • Fluid handling manifold may include fluid connections 732.
  • Fluid flow rate in the fluidics module 700 may be monitored by a flow rate sensor 734.
  • information from the flow rate sensor 734 may be provided as feedback to a fluid flow rate controller (not shown).
  • the fluid manifold 730 may be in fluid communication with the fluid handling system 150 of FIG. 1. Transport of fluids between the fluid handling system 150 and the fluidics module 700 may be accomplished using pumping mechanisms and other fluid transport devices, such as those discussed above.
  • a cell trapping chip may be housed in a cassette or cartridge, such that a needle chip may be moved into contact with the cell trapping chip
  • various implementations may include having the needle chip housed in a fluidic cartridge (e.g., a cassette) and the cell trapping chip may be configured to move to the needle chip’s location for cell engineering procedures.
  • corresponding adjustments may need to be made to the other modules to achieve equivalent functionality.
  • FIG. 8 illustrates a flow diagram of an example process for performing a cell engineering workflow according to various aspects of the present disclosure.
  • FIG. 8 illustrates a process 800 that may include a series of processes for performing cell engineering operations.
  • process 800 may include a series of operations.
  • a needle chip and a cell trapping chip are provided.
  • the needle chip and the cell trapping chip may correspond to, but not limited to, various needle chips and cell trapping chips as described herein.
  • Operation 802 may include the step of functionalizing the needle chip.
  • functionalizing the needle chips may include performing steps to prepare the needles on the needle chips to hold cargo during the workflow.
  • functionalizing the needle chips may include applying a chemical treatment to a surface of the chips.
  • functionalizing the needle chips may include applying a chemical treatment to a region of a needle chip including the needles.
  • functionalizing the needle chip may include performing a secondary treatment, non-limiting examples of which include exposing the needle chip to UV light, applying a second chemical treatment to the needle chip, and/or flowing a gas over the needle chip.
  • functionalizing the needle chip may include adding a surface chemistry to the needle chip, where the surface chemistry may enable a payload to be carried by the needles.
  • the surface chemistry may enable and/or facilitate a pay load’s adhesion to the needle chip. Additionally or alternatively, the surface chemistry may facilitate a pay load’s release into a cell.
  • functionalizing the needle chip may include storing the needle chip in a chemical solution.
  • functionalizing the needle chip may include allowing a chemical treatment to dry and/or to cure on the needle chip.
  • functionalized chips may be stored for later use. Depending on the chemical treatment applied, functionalized chips may be stored dry or wet to maintain their functionalization.
  • functionalization of the needle chip or chips may include or correspond to functionalization operations discussed below in the examples of FIGs. 9 A and 10A.
  • Operation 803 may include the step of functionalizing the cell trapping chip(s).
  • Examples of functionalizing the cell trapping chip(s) may include or correspond to various operations as described below with respect to FIGs. 9B and 10B.
  • functionalizing the cell trapping chip may include providing cell material, payload materials, buffers, or other materials to a portion or portions of the cell trapping chip.
  • Functionalizing the cell trapping chip(s) may include preparing a housing for the cell trapping chip(s) (e.g., cassette 710 as shown in FIG. 7) to receive or include cell material, payload materials, buffers, or other materials useful in performing cellular engineering applications.
  • Operation 804 may include the step of disposing the needle chip on an alignment module.
  • disposing the needle chip on an alignment module may include or correspond to operations as discussed below with respect to FIGs. 9A (e.g., with respect to operation 813) and 10A (e.g., with respect to operation 1018).
  • the alignment module may include or correspond to a portion or portions of the module assembly 200, such as, for example, the gantry stage module 220, the tip-tilt platform module 230, and/or the microscope module 240.
  • Operation 805 includes the step of aligning, by the alignment module, the needle chip with the cell trapping chip.
  • Examples of the alignment process may include or correspond to operations discussed below with respect to at least FIG. 9A (e.g., operation 819) and/or FIG. 10D.
  • alignment of the needle chip and cell trapping chip may be performed according to one or more methods as described below with respect to FIGs. 11 and 12A-12C.
  • Operation 806 may include the step of trapping a cell at a target location of the cell trapping chip by applying a pressure differential to the cell trapping chip.
  • trapping a cell at a target location of the cell trapping chip may include or correspond to functionality discussed above with respect to the fluidics module of FIG. 7.
  • trapping a cell at a target location of the cell trapping chip may include or correspond to operations discussed below with respect to FIG. 9C (e.g., operation 834) and/or FIG. 10E (e.g., operation 1052).
  • Operation 807 may include the step of impacting the cell with a needle on the needle chip by moving the needle chip toward the cell trapping chip.
  • impacting the cell with a needle on the needle chip may include or correspond to operations discussed below with respect to FIG. 9C (e.g., operation 836) and/or FIG. 10E (e.g., operation 1061).
  • Operation 808 may include the step of removing the needle chip from contacting the cell trapping chip.
  • removing the needle chip from contacting the cell trapping chip may include or correspond to, for example, operations discussed below with respect to FIG. 9C (e.g., operation 838) and/or FIG. 10E (e.g., process 1060 and/or process 1070).
  • Operation 809 may include the step of retrieving the cell from the cell trapping chip.
  • retrieving the cell from the cell trapping chip may include or correspond to functionality discussed above with respect to the fluidics module of FIG. 7. Additionally or alternatively, according to various implementations, retrieving the cell from the cell trapping chip may include or correspond to operations discussed below with respect to FIG. 9C (e.g., operation 838), FIG. 9D (e.g., operations 843, 848, and/or 851), FIG. 10E (e.g., process 1070), and/or FIG. 10F (e.g., process 1080).
  • FIG. 9C e.g., operation 838
  • FIG. 9D e.g., operations 843, 848, and/or 851
  • FIG. 10E e.g., process 1070
  • FIG. 10F e.g., process 1080.
  • FIGs. 9A-9D a flow diagram of an example process for performing a cell engineering workflow according to various aspects of the present disclosure is shown as a series of processes, including a process for loading needle chips 810, a process for loading cells 820, a process for poking cells 830, and a process for unloading cells 840.
  • FIGs. 9A-9D may include or correspond to portions of process 800 of FIG. 8. According to various aspects, FIGs. 9A-9D may illustrate an example of the processes described above with respect to FIG. 8.
  • the needle chip loading process 810 shown in FIG. 9 A may include a series of operations.
  • Operation 811 may include the step of functionalizing needle chips for use in the workflow. This functionalizing step at operation 811 may include or correspond to the functionalizing of needle chips as described in operation 802 above.
  • Functionalizing the needle chips may include performing steps to prepare the needles on the needle chips to hold cargo during the workflow.
  • Functionalizing the needle chips may include applying a chemical treatment to a surface of the chips.
  • functionalizing the needle chips may include applying a chemical treatment to a region of a needle chip including the needles.
  • functionalizing the needle chip may include performing a secondary treatment, non-limiting examples of which include exposing the needle chip to UV light, applying a second chemical treatment to the needle chip, and/or flowing a gas over the needle chip.
  • functionalizing the needle chip may include adding a surface chemistry to the needle chip, where the surface chemistry may enable a payload to be carried by the needles.
  • the surface chemistry may enable and/or facilitate a pay load’s adhesion to the needle chip. Additionally or alternatively, the surface chemistry may facilitate a pay load’s release into a cell.
  • functionalizing the needle chip may include storing the needle chip in a chemical solution.
  • functionalizing the needle chip may include allowing a chemical treatment to dry and/or to cure on the needle chip.
  • functionalized chips may be stored for later use. Depending on the chemical treatment applied, functionalized chips may be stored dry or wet to maintain their functionalization .
  • Operation 813 may include the step of installing needle chips in a component of cell engineering platform, such as, for example, needle chip holders (e.g. needle chip holder 310).
  • needle chip holders e.g. needle chip holder 310.
  • operation 815 may include the step of consolidating the needle chip holders to an organizational component on the platform, such as, for example, a hopper (e.g., hopper 320 discussed previously), a receptacle, a rest, a mount, a nest, a holder, a jacket, a chamber, a storage unit, or some other such device that can organize, receive, store, and/or hold needle chips and/or needle chip holders.
  • a hopper e.g., hopper 320 discussed previously
  • a receptacle e.g., hopper 320 discussed previously
  • a receptacle e.g., hopper 320 discussed previously
  • a receptacle e.g., a rest, a mount, a nest, a holder, a jacket, a chamber, a storage unit, or some other such device that can organize, receive, store, and/or hold needle chips and/or needle chip holders.
  • Operation 817 may include the step of selecting a needle chip for the workflow.
  • a needle chip may be selected based on its position in a needle chip holder such as have been described above with reference to FIG. 3.
  • Operation 819 may include the step of positioning the needle chip use in the work flow. Operation 819 may include securing the needle chip in its initial position in the component of the cell engineering platform. Alternatively, operation 819 may include moving the needle chip into an aligning position so that the needle chip can be co-planarized with a cell trapping chip. In various implementations, once the needle chip has been co-planarized, it may be moved into a position where it can be directly aligned with the cell trapping chip. When the needle chip is in position, the needle positioning process may be complete. Once the cell loading process 820 described below is also complete, the cell poking process 830 may be started. [0078] Referring now to FIG. 9B, FIG.
  • FIG. 9B illustrates a flow diagram of a cell loading process 820, in accordance with various implementations.
  • the cell loading process 820 may include a series of operations.
  • the operations illustrated in FIG. 9B may include or correspond to operations to functionalize the cell trapping chip.
  • functionalizing the cell trapping chip may include preparing the cell trapping chip to receive cell material.
  • Operation 821 may include the step of collecting cells in a first syringe.
  • Operation 822 may include the step of collecting a payload in a second syringe.
  • Operation 823 may include the step of installing precious materials into a cassette (e.g., a fluidics cartridge, such as cassette 710).
  • a cassette may be configured to hold precious materials and/or nonprecious materials.
  • Precious materials may include cells and/or a culture of cells in a cell culture media.
  • Precious materials may also include a genetic payload material that will be delivered to the cells. In some cases genetic payload materials (e.g. mRNA, RNP) can be expensive.
  • Precious materials may be introduced to the cassette by syringes or wells directly attached to the cassette itself in order to minimize dead volume. In some instances, precious materials may include materials with a significant cost, limited supply, and/or significant effort needed to make the material.
  • the cassette can be installed onto an alignment module. This operation may include securing the cassette in place.
  • the cassette may be installed at a predetermined location.
  • the cassette and the cell trapping chip may be primed with non-precious materials. This may include flowing a sequence of different solutions into the cassette.
  • the solutions may be transferred using fluid transportation devices, such as tubing, pumps and valves.
  • the flow rate during this step may be monitored by a flow rate sensor.
  • Operation 826 may include the step of priming the cassette with precious materials, including cell samples. Operation 826 may include flowing a cell or cells into the cassette from a syringe by applying positive pressure. [0083] In various implementations, at least a portion of the cell loading process 820 may be performed, for example, before at least a portion of the needle chip loading process 810. For example, operation 821 through at least operation 824 may be performed prior to the needle chip loading process 810 to ensure the cassette is installed before the needle chip is positioned. This may also prevent unnecessary vibrations or displacement of the needle chip after it is held in position.
  • At least a portion of the cell loading process 820 may be performed in parallel or approximately at a concurrent time as a portion of the needle chip loading process 810 (e.g., at least operations 825 and 826 may be performed concurrently with operations 811, 813, and 815).
  • FIG. 9C shows an example a cell poking process 830 according to aspects of the disclosure.
  • Operation 832 may include the step of loading precious cargo onto the needles of the needle chip.
  • the precious cargo may include materials to be inserted into a cell (e.g., payload materials, genetic materials, pharmaceutical materials, and/or chemical materials).
  • a vacuum may be applied to the cell trapping chip.
  • a vacuum may be applied to attract cells toward target locations as discussed previously. These target positions can be pores in the cell trapping chip. In various implementations, pores can be dimensioned with a smaller diameter than the cells. Thus, the cells may become trapped at the pores.
  • applying a vacuum to the cell trapping chip may form a microfluidic channel between the microneedle chip and the cell trapping chip.
  • the vacuum may be applied when the needle chip reaches a preset height above the pores on the cell trapping chip.
  • the vacuum may be a hydraulic pressure difference in the fluid between one side of the cell trapping chip and the other side.
  • a low magnification microscope may scan over the cell trapping chip to determine the cell trapping efficiency.
  • the user will input the desired poking parameters (e.g., approaching velocity, insertion distance, and/or needle in-cytoplasm duration). Inputting the desired parameters may cause a controller to adjust the vacuum level on the chips, add fluid to the system, introduce secondary stage materials to the microfluidic interface, and/or adjust a timer for a system process.
  • the relevant parameters may be preprogrammed into the system controller based on a particular cell engineering workflow.
  • Operation 836 may include the step of poking the cells with the needles on the needle chip. This operation may include causing the microneedles to contact, impact, or poke into the cells.
  • the cell poking may be performed according to the cell poking parameters.
  • a video feed may be configured to record data during the process.
  • sensors may be configured to record data during the process.
  • data from the video feed and/or sensor may be saved in storage (e.g., a computer memory).
  • data may be transmitted remotely to a storage device (e.g., a cloud server).
  • Operation 838 may include the steps of releasing the vacuum and recapturing the cells.
  • releasing the cell trapping chip from the vacuum may include applying positive pressure to the cell trapping chip for a preset time duration using a pump.
  • the pump may include a flow rate sensor configured to provide flow rate feedback to the system.
  • Recapturing the cells may include collecting the cells to a retrieval chamber in the cassette. For example, a syringe may be used to provide the necessary pressure to move the processed cells into the retrieval chamber.
  • FIG. 9D shows an example cell unloading process 840.
  • Cell unloading process 840 may include a series of determinations and operations, examples of which are outlined below.
  • determination 841 of the cell unloading process 840 a determination may be made as to whether a cycle of a cell engineering process is complete. If the cycle of the cell engineering process is complete, then at operation 843 cells may be collected. Collecting cells may include flowing the cells out of the cassette. This may be done, for example, using a syringe. Collecting cells may be alternatively performed by a pumping system. The cells may then be collected into a container.
  • poked and unpoked cells may be flowed into respective dedicated output reservoirs (e.g., chambers or cavities) in the fluidics cartridge such that at the end of the poking operation, users can retrieve cells from those reservoirs.
  • a fluidics cartridge may be used for multiple operations (e.g., multiple batches of the same cell engineering process). It may be desirable in such circumstances to have cell samples stored in a reservoir so they may all be retrieved at one time.
  • a determination 844 may include determining whether a new cycle is to be performed. If a new cycle is to be performed, then the cell unloading process 840 may end and the new cycle may be started from operation 823 of FIG. 9B. In that case, in various instances, time to install a new cassette may be provided. If, at determination 841 of FIG. 9D, a new cycle is not to be performed, then at operation 845 the cell engineering process may completed, and a shutdown procedure may be initiated.
  • a determination 842 may be made as to whether to reuse the cassette. If the cassette is not to be reused, the process may go to operation 843 and follow the same flow as if the cycle were complete, as described above.
  • a determination 846 may be made as to whether to reuse the needle chip. If the needle chip is to be reused, then a determination 847 may be made as to whether to collect cells. In various implementations it may be inefficient to collect all the cells each time the process is run, and so cell collection may be preferably delayed. If cells are to be collected, then at operation 848 the cells may be collected. Operation 848 may be functionally equivalent to operation 843 described above. After the cells are collected at operation 848, or if the determination 847 cells were not to be collected, then at operation 849 the old needle chip may be returned to the location it was retrieved from. The cell unloading process 840 would then end, and needle chip loading process 810 of FIG. 9A may be rejoined at operation 817, where a new needle chip would be selected.
  • a determination 850 may be made as to whether to collect cells. If cells are to be collected, then at operation 851 the cells may be collected. Operation 851 may be functionally equivalent to operation 843 described above.
  • a determination 852 may be made as to whether the needle chip is to be cleaned. If the needle chip is to be cleaned, then the process flows to operation 853. Operation 853 may include the step of cleaning the needle chip. In various implementations cleaning the needle chip may include washing the needle chip with ethanol, and then with phosphate buffered saline (PBS) or some other solvent compatible with the needle cargo.
  • PBS phosphate buffered saline
  • FIGs. 10A through 10G illustrate flow diagrams of an example process for performing a cell engineering workflow according to aspects of the present disclosure. Portions of FIGs. 10A to 10G may correspond to and/or overlap with portions of FIGs. 9 A through 9D.
  • Cell engineering processes, steps, and subprocesses illustrated in FIGs. 10A to 10G may include the processes and steps as presented in FIGs. 10A to 10G, or a subset of the processes and steps as presented in FIGs. 10A to 10G. In various implementations, the processes and steps may be performed in a different order to the extent that such reordering is reasonable. Some of the processes, routines, subroutines, and steps may be optional and may not need to be performed in every instance.
  • FIGs. 10A through 10G illustrate flow diagrams of an example process for performing a cell engineering workflow according to various aspects of the present disclosure.
  • the flow diagrams of FIGs. 10A through 10G may include or correspond to methods or portions of methods discussed herein, including with respect to FIG. 8 and FIGs. 9A-9D.
  • the flow diagrams of FIGs. 10A through 10G may include several processes and/or subprocesses, including, for example, 1) a needle chip preparation process 1010, 2) a cassette assembly process 1020, 3) a needle chip and cartridge loading process 1030, 4) a chip planes synchronization and final XY alignment process 1040, 5) a cell capture and payload loading process 1050, 6) a cell poking process 1060, 7) a cell release and collection process 1070, 8) a post poke cell handling process 1080, and 9) a system shutdown process 1090.
  • a needle chip preparation process 1010 2) a cassette assembly process 1020, 3) a needle chip and cartridge loading process 1030
  • 4) a chip planes synchronization and final XY alignment process 1040 a cell capture and payload loading process 1050, 6) a cell poking process 1060, 7) a cell release and collection process 1070, 8) a post poke cell handling process 1080, and 9) a system shutdown process 1090.
  • a needle chip preparation process 1010 may include various subprocesses and/or operations. According to various operations, the needle chip preparation process 1010 may include operations to functionalize the needle chips.
  • the system may be configured to accept delivery of a needle chip or a plurality of needle chips.
  • the needle chips may be formed in a large batch and/or attached to each other, such as may be the case, for example, when needle chips are formed on and/or from a silicon substrate (e.g., a silicon wafer), on or from other semiconductor materials such as gallium arsenide, or on and/or from other substrates or materials suitable for microfabrication of needle chips.
  • the needle chip(s) may be broken into individual dice of a single chip or multiple needle chips.
  • needle chip wafers may be scribed and broken, mechanically sawn (e.g., with a dicing saw), or cut with a laser cutter. This process may also be referred to as die singulation.
  • the surface of the needle chip(s) may be cleaned.
  • cleaning the surface may include dipping and/or spraying operations to remove impurities.
  • dipping may include dipping the wafer, dice, and/or individual needle chip(s) into a chemical solution or ultra-pure deionized water.
  • spraying may include spraying a chemical in liquid or gas form onto a spinning wafer or onto individual chips.
  • surface plasma activation may be performed.
  • surface plasma activation may include sputtering with an inert gas and/or etching with chemically reactive plasma.
  • the chip(s) may be immersed in a thiol solution. The immersion .
  • the chip(s) may be removed from the thiol solution and rinsed in an appropriate rinsing material.
  • the chip(s) may be dried and stored.
  • the needle chip(s) may be mounted into needle chip holders, such as, for example needle chip holders 310 as have been described herein.
  • the cassette assembly process 1010 may include operations to functionalize cell trapping chips and/or cassettes.
  • the cell trapping chip(s) e.g., vacuum traps
  • the cell trapping chips may be singulated (e.g., separated into dice of individual chips or multiple chips).
  • the cell trapping chips may be separated from each other by being scribed and broken, mechanically sawn (e.g., with a dicing saw), or cut with a laser cutter.
  • Operation 1021 may also include plasma treating the cell trapping 1 chip(s).
  • the cell trapping chip(s) may be treated by sputtering the cell trapping chip(s) with an inert gas and/or etching with chemically reactive plasma.
  • the cell trapping chip(s) e.g., vacuum trap(s)
  • the process may include a step for loading priming syringes into the cassette and priming the syringe connection.
  • cells e.g., cells in a cell solution
  • Cell syringes may be distinct from priming syringes.
  • the priming syringes may be swapped for (e.g., removed and replaced by) cell syringe(s) and payload syringe(s).
  • Payload syringes may be prepared to include payload materials (e.g., genetic payloads), such as have been discussed herein.
  • FIG. 10C a flow diagram of a needle chip and cartridge loading process 1030 is illustrated.
  • a system check may be performed.
  • a system initialization may be performed.
  • needle chip(s) may be mounted on to the system.
  • the needle chips may be functionalized and provided to operation 1033 from the output of operation 1018.
  • the needle chip(s) may be mounted in needle chip holders.
  • mounting needle chip(s) on to the system may include mounting a chip holder or needle chip holders in wells in a needle chip hopper or other organizational component as has been described herein. Additionally or alternatively, mounting needle chip(s) on to the system may include mounting the organizational component to the system.
  • the cassette may be loaded onto the system.
  • the cassette may be prepared for mounting from the output of operation 1025.
  • the cassette may be secured in place by clamps or other securing mechanisms (e.g., securing member 518).
  • the cassette may be primed.
  • Operation 1036 may include moving (e.g., picking up) a needle chip holder with a robotic gripper (e.g., robotic gripper 340 described above).
  • the synchronization and final XY alignment process 1040 may be configured to align the needle chip with the cell trapping chip (or, more precisely, to align a needle on the needle trapping chip with target locations at which cells may become trapped in the cell trapping chip).
  • the location of the vacuum chip (e.g., cell trapping chip) and the location of the needle chip may be determined.
  • the locations of the cell trapping chip and needle chip may be precisely determined according to methods described herein (e.g., with reference to the functionality described relative to the sensors 332 in FIG.
  • the chips may be translated to a pre-insertion XY position.
  • the pre-insertion XY position may include or correspond to an aperture in the cassette corresponding to the cell trapping chip.
  • the needle holder may be configured to be insertable into the aperture in the cassette.
  • the needle chip (and the needle chip holder) may be inserted into the cassette.
  • a diaphragm seal may be engaged. The diaphragm seal may be engaged until it is within a specified value at determination 1046. When the diaphragm seal is within a specified value, at operation 1047 the needle chip may be translated in a Z direction to its prepoke distance.
  • a final XY translation may be performed to complete the alignment process.
  • FIG. 10E flow diagrams of a cell capture and payload loading process 1050, a poking process 1060, and a release and collection process 1070 are illustrated.
  • a payload may be loaded into or onto the needles of the needle chip.
  • cells may be flowed into the cell trapping chip (e.g., from the cassette) and trapped at target locations in the cell trapping chip (e.g., by applying a pressure differential to the cell trapping chip).
  • the system may cause the needles of the needle chips to poke the cells trapped at the target locations of the cell trapping chip.
  • the system may be configured to perform the poking process automatically.
  • the poking process may include process steps to retract needles from the cell trapping chip after the poking has been accomplished.
  • parameters may be input and/or captured during the process.
  • the poking process may be configured based on parameters such as a descent velocity, a starting position, a poking distance (e.g., a distance into a cell that a needle may be poked), a residence time (e.g. how long the needle remains in the cell), an ascent velocity, and/or a number of times the cell is to be poked.
  • Other parameters may be factored into the poking process, and not all parameters need to be measured and/or input for the process to be performed.
  • the flow on the bottom of the cassette may be reversed. This may cause cells to flow into an output reservoir in the cassette.
  • cells may be collected on or in a cassette well (e.g., the output reservoir).
  • the needle chip may be retracted.
  • the cells may be removed from the cassette well (e.g., the output reservoir). For example, the cells may be removed using a pipette, or the cells may be removed through some other method of transporting cell materials and/or cell solutions.
  • the cells may be provided for operation 1081 as described below.
  • the cell trapping chip e.g., vacuum trap
  • the cells may be moved (e.g., using a pipette) into a plate.
  • the cells may be moved into a well or wells of a well plate, such as a 6-well plate, a 24-well plate, a 48-well plate, a 96-well plate, a 384 well plate, a 1536-well plate, a 3456-well plate, or a 9600-well plate.
  • a well plate is described here only by example, as the cells may be moved into another appropriate location at this step.
  • the cell media may be modified (e.g., to facilitate analysis of the cells).
  • the cell or cells may be plated into an appropriate well plate. This may include a different well of the well plate described above, or a different well plate.
  • the cell health may be analyzed. For example, a cell count may be performed. Additionally or alternatively, cell viability may be determined. For example, cell viability may be calculated using a ratio of the total living cells to the total number of cells in the sample.
  • analysis may be performed on the cells and on cargo delivered to the cells. For example, changes in the cell viability may be analyzed. This analysis may take place over an extended period of time, such as over a day or a number of days.
  • FIG. 10G a flow diagram of a system shutdown process 1090 is illustrated.
  • the needle chip may be removed and mounted on a microscope fixture.
  • the microscope fixture may include or correspond to the microscope module 600 described herein, or it may include some other microscope fixture.
  • the needle chip may be imaged off system.
  • the cartridge may be disassembled and disposed of.
  • the system may enter a system shut-down routine.
  • the system, or individual system components may be cleaned and/or decontaminated as needed.
  • some of the processes of FIGs. 10A-10G can be run in parallel. Processes that may run in parallel may start at the same time, or may be configured to start at different times.
  • the needle chip preparation process 1010 of FIG. 10A, the cassette assembly process 1020 of FIG. 10B, and the needle chip and cartridge loading process 1030 of FIG. 10C may be configured to run at least partially in parallel and provide inputs and/or outputs to each other, such as is shown in FIGs. 10A-10C.
  • processes may end at different times, disregarding which of the processes finish first.
  • the post poke cell handling process and the system shutdown process may finish at different times, as shown in FIGs. 10F and 10G.
  • FIGs. 10A-10G may be performed by users or by automated systems. Any indication of whether a process is performed by a user or an automated system is included as an example and is not intended to limit the manner in which a process may be performed.
  • FIGs. 10F and 10G highlight examples of cell handling and system shut down processes that may be optional to the method.
  • analyzing cell health or analyzing cargo as in FIG. 10E may not need to be performed in every instance, or as part of the same process flow.
  • the timing indicated for the analysis in the last step of post poke cell handling process 8 in FIG. 10E may be performed within a few days, but is not limited to this time frame. In various instances the analysis may be performed momentarily after the cell poking process, and in other instances more time such as days later may be needed for proper analysis. In any case, the time frame is given as an example and not as a limitation. In FIG.
  • shutting down the system after each time the process is performed may not be necessary, such as, for example, in instances when multiple processes may be performed concurrently using the same needle chip and/or cell trapping chip, or with new needle chips and/or cell trapping chips without shutting down the system completely.
  • FIGs. 11A-11D illustrate an example of aspects of the needle chip loading process 810 and the cell loading process 820 as applied to a system in accordance with various implementations, including, for example, system 100.
  • a user may load a cassette 710 into the cassette socket 514 on the platform 510.
  • the cassette 710 may be secured in place with securing member 518 and fastened in place with fastener 519.
  • FIG. 11A may include or correspond to operation 824 of cell loading process 820.
  • the cassette 710 may be primed with fluids, including, for example, any number of a cell sample culture, a buffer, and/or a reagent. This may occur before the cassette 710 is loaded onto the platform 510, or after the cassette is loaded onto the platform 510.
  • the hopper 320 loaded with needle chip holders 310 may be loaded onto the hopper rest 330 on platform 510. This may include or correspond to operation 815 of needle chip loading process 810 described above with reference to FIG. 9A.
  • the hopper 320 may be installed such that the needle chip holders 310 are located at predefined known positions.
  • the system may detect the number of needle chips being loaded.
  • the hopper 320 may be configured to allow the system 100 to determine the position of all the needle chips beforehand, for example by using the sensors 332 in the hopper rest 330. When the positions of the needle chips have been determined, the robotic gripper 340 may move to the needle chip position and picks it up as shown in FIG. 11B. This process may include or correspond to operation 817 of needle chip loading process 810.
  • FIGs. 11C-11D may include or correspond to operation 819 of needle chip loading process 810.
  • the robotic gripper 340 gripping the needle chip holder 310 may be moved using the gantry stage module 400 (can also be referred to as a gantry module) to a first aperture 512 in the platform 510.
  • the system may estimate the rough location of the needle chip using the low magnification microscope (e.g., low magnification microscope 640 of FIG. 6A) and a focus algorithm, such as a predictive focus algorithm.
  • the system may also determine the precise focus plane of the needle chip and the cell trapping chip 718 using the high magnification microscope (e.g., high magnification microscope 650 of FIG. 6A).
  • an image processing routine may be used to determine the precise location of the needle chip and cell trapping chip 718.
  • the gantry stage module 400 and the tip-tilt platform module 500 may adjust the positions of the needle chip and the cell trapping chip 718 to bring them into the same or a substantially parallel plane.
  • the system may move the needle chip into the cassette 710 through opening 716 and into contact with the cell trapping chip 718.
  • FIGs. 11A-11D may be performed by users or by automated systems. Any indication of whether a process is performed by a user or an automated system is included as an example and is not intended to limit the manner in which a process may be performed.
  • An alignment process for a cell engineering workflow may include estimating the coordinates of features of a needle chip, estimating the coordinates of features of a cell trapping chip, and based on the estimated coordinates of the features of the needle chip and the estimated coordinates of the features of the cell trapping chip, moving at least one of the needle chip and the cell trapping chip into alignment with the other chip. After the alignment process is complete, the needle chip and the cell trapping chip will have surfaces aligned into the same plane.
  • the chips to be aligned may be a microneedle chip and a cell trapping chip.
  • the needle trapping chip may be formed with the same pitch of features and the same alignment fiducials as the needle chip. Chips formed in this manner can enable the use of optical alignment systems and robotics to align every nanoneedle to every cell trap.
  • the chips may be co-planarized to within 1 pm of error in four edges. In order to ensure every nanoneedle on the needle chip pokes into a single cell at a target location (e.g in pores) in the cell trapping chip, careful alignment is required. However, the nanoneedles on the chips may be fragile, and thus cannot afford to perform contact alignment. Cell trapping chips may also be too fragile for contact alignment.
  • Chips designed for contact alignment would need a more complex mechanical design to protect the nanoneedles. Instead, alignments may be made using a microscope to locate key features of the chips in three-dimensional space and a tip-tilt platform to move one of the chips into alignment with the other chips as described below. Alignments made using a tip-tilt platform may be confirmed optically before cell engineering processes on the chips are begun. In various implementations, at least one of the cell trapping chip and the needle chip may be formed of a transparent material. This may facilitate effective alignment between the chips.
  • FIGs. 12A to 12C show a diagram of an example alignment process according to aspects of the present disclosure.
  • FIGs. 12A-12C include illustrations of stages of an alignment process 1200.
  • a high magnification microscope 1220 estimates the coordinates of key features of the needle chip 1210 using a sample of data points.
  • the key features may include, for example, a first edge 1212 of the needle chip 1210, a second edge 1214 of the needle chip 1210, corners of the needle chip 1210, and/or fiducial marks located on the needle chip 1210.
  • the coordinates of key features may first be roughly estimated by a low magnification microscope (not shown).
  • a high magnification microscope 1220 can provide a narrow field of view. Using a low magnification microscope in a first stage can provide a wider field of view for an initial coordinate estimate.
  • the coordinate estimate may correspond to the position of three of the edges of the needle chip. Identifying, estimating, and/or locating the unique coordinates of three key features of the needle chip 1210 may define a plane in which the needle chip 1210 is located. In various implementations, the coordinate estimate may correspond to the location of fiducial marks. Such fiducial marks may be located at the edge of the needle chip 1210. Again, the coordinates of at least 3 fiducial marks may be needed to define the plane of the needle chip 1210.
  • the coordinates may correspond to a coordinate vector in three dimensional space.
  • coordinates may be estimated with reference to a cartesian coordinate system (e.g., X, Y, and Z coordinates). Additionally or alternatively, the coordinates may be estimated with reference to angles of rotation. For example, in FIG. 12A the coordinates of an edge 1212 of the needle chip 1210 are suggested by reference to an offset Az, an x plane, and an angle 9 between the needle chip 1210 and the x plane.
  • the coordinate information may be stored, for example in a computer memory, for use in alignment algorithms as described with respect to FIG. 12C below.
  • the alignment process may use a microscope to accurately focus on fiducial marks that may be located at the corners of the needle chip and cell trapping chip.
  • a predictive focus algorithm may be used to help locate a focus plane of the chip. Examples of a predictive focus algorithm will be discussed in further detail below.
  • Identifying, estimating, and/or locating the unique coordinates of three key features of the needle chip 1210 may define a plane in which the needle chip 1210 is located.
  • the coordinates of one of the key features of the needle chip 1210 are estimated, the coordinates of at least two other separate key features of the needle chip 1210 may be estimated in a similar manner.
  • the coordinates of a first fiducial on a first corner (e.g., the top-left comer) of the needle chip 1210 may be determined and subsequently recorded.
  • the coordinates of a second fiducial on a second comer e.g., the top-right comer
  • a third fiducial on a third corner (e.g., the bottom-right corner) of the needle chip 1210 may be determined and subsequently recorded.
  • This example is intended to be illustrative and not limiting, and it is anticipated that any of several alternate key features or locations on a needle chip may be used to identify the a plane in which the needle chip may be located, just as any of several alternative orders in which a needle chip’s key features are located may be used.
  • the microscope 1220 may be used to estimate the coordinates of the key features of the cell trapping chip 1240. This may be done in a similar manner to estimating the coordinates of key features of the needle chip 1210.
  • the coordinates of key features of the cell trapping chip 1240 may be estimated using a high magnification microscope 1220 or multiple microscopes (e.g. a low magnification microscope and high magnification microscope 1220). The coordinates may be defined using the same coordinate system as used for key features of the needle chip 1210 for ease of comparing the coordinates of the cell trapping chip 1240 to the coordinates of the needle chip 1210.
  • the estimated coordinates of three separate key features of the cell trapping chip 1240 may be estimated in a similar manner to the three separate key features of the needle chip 1210.
  • the three key features of the cell trapping chip 1240 may correspond to the three key features of the needle chip.
  • FIG. 12C illustrates an example of aligning the cell trapping chip 1240 into planar alignment with needle chip 1210 by adjusting the angle of cell trapping chip 1240.
  • estimating and/or locating the unique coordinates of three key features of the needle chip 1210 may define a plane in which the needle chip 1210 is located.
  • Estimating and/or locating the unique coordinates of three key features of the cell trapping chip 1240 may define a plane in which the cell trapping chip 1240 is located.
  • Alignment processes as discussed here in reference to stage c) may function to bring the cell trapping chip 1240 into the same plane as the needle chip 1210.
  • an alignment algorithm may be used to compute the necessary movement parameters to align the chips based on the estimated coordinates of the needle chip 1210 and the cell trapping chip 1240.
  • the computed movement parameters may correspond to an offset between the coordinate vectors of the needle chip 1210 and the corresponding coordinate vectors of the cell trapping chip 1240.
  • the coordinate vectors from the key features on each of the chips may form a pair of coordinate vectors.
  • the correction angles needed may be computed. These corrections may then be converted to the position of motorized stages of a tip-tilt platform.
  • a tiptilt platform may be used to move at least one of the cell trapping chip 1240 and the needle chip 1210 into a calculated orientation such that the chips are aligned into the same plane.
  • the goal of the alignment process is to manipulate the needle chip so that the separation distance between at least three corners of the cell trapping chip 1240 and at least three corresponding corners of the needle chip 1210 is the same. If the separation distance is the same between the corresponding comers of the chips is the same for each corner, that implies that the chips are planarized.
  • needle chips and cell trapping chips may be aligned using an alignment algorithm.
  • the alignment algorithm may include steps of identifying the coordinates of key features of a needle chip, identifying the coordinates of key features of a cell trapping chip, determining the plane in which the needle chip is positioned, determining the plane in which the cell trapping chip is positioned, determining a misalignment between the plane of the cell trapping chip and the plane of the needle chip, calculating the movement of one of the chips to bring the chip into the plane of the other chip, and moving the chip into alignment.
  • a needle chip or a cell trapping chip may be disposed on a tip-tilt platform (e.g., an alignment module).
  • the chip that is not disposed on the tip tilt-platform may be held in a fixed position relative to the other chip.
  • a tip-tilt platform may include pushers (e.g., linear stages).
  • the pushers may be arranged at comers of the tip-tilt platform.
  • the pushers may include motors.
  • the motors may be connected to a controller. The controller may be configured to run the motors and extend and/or retract the pushers.
  • Extending and/or retracting the pushers may move the tip-tilt platform and/or alter its pitch and/or roll angle relative to the fixed position of the other chip.
  • the controller may be configured using a transformation.
  • the transformation may translate a calculated plane of alignment into a calculated set of pusher positions. Moving the pushers to the calculated set of pusher positions may bring the chip on the tip-tilt platform into the calculated plane of alignment.
  • a microscope with a high magnification objective lens may be used to focus on fiducial markings on both the microneedle chip and the cell-trapping chip.
  • a high magnification microscope can desirably create a shallow depth of field to determine the accurate focus plane.
  • a high magnification microscope gives a narrow field of view. But the narrow field of view can be mitigated by the following exemplary predictive focus algorithm.
  • the predictive focus algorithm may have 4 stages: image acquisition, image analysis, optimal focus determination, and focus adjustment. The stages are described below. [0135] (1) Image acquisition.
  • the high magnification microscope captures a series of images at different focal planes (or Z-positions) within a predefined range above and below an initial focal plane.
  • the range of focal planes will contain the focal plane at which the chip (either the microneedle chip or the cell-trapping chip) is located.
  • the images may be analyzed to determine the sharpness and/or contrast within each image.
  • Several methods can be used to analyze image sharpness, such as, for example, calculating the variance, sum-modified Laplacian, or Tenengrad function of the image.
  • the microscope's hardware components such as, for example a motorized stage or a focusing mechanism, moves the objective lens or the stage to the optimal focal plane as determined by the algorithm.
  • the predictive focus algorithm may be configured to focus its image analysis on the key features of the needle chip 1210 or the cell trapping chip 1240.
  • Alignment processes such as those described herein may enable precision automation of cell engineering workflows.
  • Precision automation may provide several benefits to cell engineering workflows and applications. For example, at least the following benefits may be achieved.
  • Consistency and reproducibility Automated systems can precisely follow predefined protocols, ensuring consistent results with minimal variability. This leads to improved reproducibility.
  • Increased throughput Automation allows for the simultaneous handling of multiple samples, increasing the overall throughput and efficiency of the process. This may be particularly valuable when working with large sample numbers or high-throughput screening applications.
  • Reduced human error Automated systems minimize the risk of human error, such as pipetting mistakes or inconsistencies in handling, which can compromise the quality of the transfection process and the subsequent data.
  • Enhanced safety Some transfection methods involve the use of hazardous materials or biohazardous samples. Automated systems can reduce the risk of exposure to these materials, ensuring a safer working environment.
  • FIG. 13 an example of a computing device that is operable to support cell engineering workflows and methods according to one or more aspects of the present disclosure is shown as a computing environment 1300 that includes a computing device 1310.
  • the computing device 1310 may be operable to initiate or control cell engineering workflows including chip alignment processes or other cell engineering workflows of the stages of any of the processes described with reference to, for example, FIGs. 8-12.
  • the computing device 1310 includes at least one processor 1320 and system memory 1330.
  • the system memory 1330 may be volatile (such as random access memory or “RAM”), non-volatile (such as read-only memory or “ROM,” flash memory, and similar memory devices that maintain stored data even when power is not provided) or some combination of the two.
  • the system memory 1330 typically includes instructions 1332 and one or more applications.
  • the at least one processor 1320 may be operable to execute the instructions 1332 to perform one or more operations described herein, including, for example, operations of the method 810 of FIG. 9A, the method 820 of FIG. 9B, the method 830 of FIG. 9C, the method 840 of FIG. 9D, the method 900 of FIG.
  • the instructions 1332, the applications, or both may be located at multiple computing devices, where the multiple computing devices are part of a distributed computing system.
  • one or more of the multiple computing devices of the distributed system may comprise the representative computing device 1310.
  • the computing device 1310 may also have additional features or functionality.
  • the computing device 1310 may also include removable and/or non-removable data storage devices such as magnetic disks, optical disks, tape, and standard- sized or miniature flash memory cards.
  • Such additional storage is illustrated in FIG. 13 by storage 1340.
  • Computer storage media may include volatile and/or non-volatile storage and removable and/or non-removable media implemented in any method or technology for storage of information such as computer- readable instructions, data structures, program components or other data.
  • the system memory 1330 and the storage 1340 are examples of computer storage media.
  • the computer storage media may include, but is not limited to, RAM, ROM, electrically erasable programmable read-only memory (EEPROM), flash memory or other memory technology, compact disks (CD), digital versatile disks (DVD) or other optical storage, magnetic cassettes, magnetic tape, magnetic disk storage or other magnetic storage devices, or any other medium that can be used to store information and that can be accessed by computing device 1310. Any such computer storage media may be part of the computing device 1310.
  • the computing device 1310 may also have input/output (VO) device(s) 1350, which may include input devices, such as a keyboard, mouse, pen, voice input device, touch input device, etc., output device(s), such as a display, speakers, a printer, etc., or a combination thereof.
  • VO input/output
  • the computing device 1310 also contains one or more communication interface(s) 1360 that allow the computing device 1310 to communicate with a cell engineering system 1380 via a wired or a wireless network 1370.
  • the cell engineering system 1380 may include one or more cell engineering process modules, one or more computing devices, cell engineering tools or devices, or a combination thereof.
  • the cell engineering system 1380 may initiate or facilitate, for example, any of the stages or operations of the processes described with reference to FIGs. 9A-9D, FIGs. 10A-10G, FIGs. 11A-1 ID, or FIGs. 12A-12C.
  • the communication interface(s) 1360 are an example of communication media.
  • communication media may include wired media such as a wired network or direct-wired connection, and wireless media, such as acoustic, radio frequency (RF), infrared and other wireless media.
  • RF radio frequency
  • the VO device(s) 1350 may be optional.
  • Components, the functional blocks, and the modules described herein include processors, electronics devices, hardware devices, electronics components, logical circuits, memories, software codes, firmware codes, among other examples, or any combination thereof.
  • features discussed herein may be implemented via specialized processor circuitry, via executable instructions, or combinations thereof.
  • the hardware and data processing apparatus used to implement the various illustrative logics, logical blocks, modules, and circuits described in connection with the aspects disclosed herein may be implemented or performed with a general purpose single- or multi-chip processor, a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic device, discrete gate or transistor logic, discrete hardware components, or any combination thereof designed to perform the functions described herein.
  • a general purpose processor may be a microprocessor, or any conventional processor, controller, microcontroller, or state machine.
  • a processor may also be implemented as a combination of computing devices, such as a combination of a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors in conjunction with a DSP core, or any other such configuration.
  • particular processes and methods may be performed by circuitry that is specific to a given function.
  • the functions described may be implemented in hardware, digital electronic circuitry, computer software, firmware, including the structures disclosed in this specification and their structural equivalents thereof, or any combination thereof. Implementations of the subject matter described in this specification also may be implemented as one or more computer programs, that is one or more modules of computer program instructions, encoded on a computer storage media for execution by, or to control the operation of, data processing apparatus.
  • the functions may be stored on or transmitted over as one or more instructions or code on a computer-readable medium.
  • the processes of a method or algorithm disclosed herein may be implemented in a processor-executable software module which may reside on a computer-readable medium.
  • Computer-readable media includes both computer storage media and communication media including any medium that may be enabled to transfer a computer program from one place to another.
  • a storage media may be any available media that may be accessed by a computer.
  • Such computer- readable media can include random-access memory (RAM), read-only memory (ROM), electrically erasable programmable read-only memory (EEPROM), CD-ROM or other optical disk storage, magnetic disk storage or other magnetic storage devices, or any other medium that may be used to store desired program code in the form of instructions or data structures and that may be accessed by a computer. Also, any connection may be properly termed a computer-readable medium.
  • Disk and disc includes compact disc (CD), laser disc, optical disc, digital versatile disc (DVD), floppy disk, hard disk, solid state disk, and Blu-ray disc where disks usually reproduce data magnetically, while discs reproduce data optically with lasers.
  • an ordinal term e.g., “first,” “second,” “third,” etc.
  • an element such as a structure, a component, an operation, etc.
  • an ordinal term does not by itself indicate any priority or order of the element with respect to another element, but rather merely distinguishes the element from another element having a same name (but for use of the ordinal term).
  • Coupled is defined as connected, although not necessarily directly, and not necessarily mechanically; two items that are “coupled” may be unitary with each other.
  • compositions when used in a list of two or more items, means that any one of the listed items may be employed by itself, or any combination of two or more of the listed items may be employed. For example, if a composition is described as containing components A, B, or C, the composition may contain A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.
  • A, B, and/or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.
  • “and/or” operates as an inclusive “or.”
  • substantially is defined as largely but not necessarily wholly what is specified - and includes what is specified; e.g., substantially 90 degrees includes 90 degrees and substantially parallel includes parallel - as understood by a person of ordinary skill in the art.
  • the term “substantially” may be substituted with “within [a percentage] of’ what is specified, where the percentage includes 0.1, 1, 5, and 10 percent; and the term “approximately” may be substituted with “within 10 percent of’ what is specified.

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Abstract

Système d'administration de macromolécules ou d'autres matériaux dans des cellules pouvant comprendre des micro-aiguilles formées sur des puces à micro-aiguilles, des puces de piégeage de cellules, et des systèmes d'alignement de puces à micro-aiguilles avec des puces de piégeage de cellules. Selon divers modes de réalisation, des systèmes peuvent comprendre des appareils pour manipuler des puces à micro-aiguilles, un portique, un module d'alignement, un module de microscope et un module fluidique. L'invention concerne également des procédés d'alignement des puces de piégeage de cellules et des puces de micro-aiguilles et des procédés de réalisation de processus d'ingénierie cellulaire à l'aide des systèmes.
PCT/US2024/034800 2023-06-21 2024-06-20 Système intégré de distribution de macromolécules dans des cellules individuelles à l'aide d'une commande et d'une automatisation de mouvement précis Pending WO2024263765A2 (fr)

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US7270730B2 (en) * 2000-08-04 2007-09-18 Essen Instruments, Inc. High-throughput electrophysiological measurement system
GB0502556D0 (en) * 2005-02-08 2005-03-16 Lab901 Ltd Analysis instrument
JP5099688B2 (ja) * 2007-11-13 2012-12-19 Ｎｔｎ株式会社 液状材料塗布装置およびそれを用いた欠陥修正装置
EP2344893B1 (fr) * 2008-09-16 2014-10-15 Ibis Biosciences, Inc. Systèmes et procédés de manipulation de microplaques
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