Loading station
Inventors
Lai, Janice H. • Spuhler, Philipp S. • Facer, Geoffrey R. • Li, Sixing • Cesar, Christopher G.
Assignees
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Publication Number
US-11061043-B2
Publication Date
2021-07-13
Expiration Date
Abstract
Disclosed herein are methods, devices, and systems for loading and retrieval of particles. In some embodiments, a loading station comprise a tray configured to receive a microwell array, a first magnet, a second magnet, and an actuation mechanism configured to cause movement of at least one of the first magnet and the second magnet.
Core Innovation
A loading station is provided for single-cell stochastic barcoding workflows, including a tray that receives a cartridge with a flowcell and a microwell array. The tray positions the cartridge in the loading station so that fluids are introduced via an inlet port and delivered through a fluidic channel over the microwells, and so that outflow occurs via an outlet port. The loading station includes a magnetic-based arrangement for controlling magnetic nucleic acid barcode-bearing beads or magnetic barcode-bearing beads within the microwells.
The loading station includes a first magnet and a second magnet that are movable by an actuation mechanism between active and inactive positions. The magnets are arranged so that they apply magnetic forces in different directions, including a first direction and a second direction different from the first direction. The magnets have spatial relationships to the flowcell characterized as inferior and superior spacing constraints, and mechanical coupling ensures only one magnet is active at a time.
The loading station further includes a drawer and tube receptacle arrangement for routing waste versus bead collection, with positions such as a waste collection position and a bead collection position. Cells and barcode-bearing beads are introduced into the microwells, lysis is performed while holding beads using the first magnet, beads are retrieved using the second magnet, cells are washed away, and then beads are released and flushed out for collection through a collection tube.
Claims Coverage
The partial content provides two independent method claims that cover bead retention and directionally controlled bead movement using two magnets in different force directions, and bead maintenance, cell lysis, and subsequent bead release and flush using first and second magnets positioned inferior and superior to the flow cell, respectively. Each independent claim is characterized by multiple inventive features focused on cartridge flowcell microwells, magnetic barcode-bearing beads, and magnet positioning and force direction control to move beads out of the cartridge.
Fluid introduction into a microwell flowcell cartridge with cells
introducing a fluid comprising a plurality of cells into a flowcell of a cartridge positioned within a loading station, wherein the flowcell comprises a fluidic channel having a bottom comprising a plurality of microwells, and wherein the liquid is introduced into the inlet port and flowed across the plurality of microwells so that cells are positioned in the microwells
Introducing magnetic barcode-bearing beads sized for microwells
introducing a fluid comprising a plurality of magnetic barcode-bearing beads into the flowcell, wherein each microwell is dimensioned to receive at least one cell and at least one magnetic barcode-bearing bead, and wherein the fluid is introduced into the inlet port and flowed across the plurality of microwells
Two magnets exert different-direction magnetic forces to control bead flow
moving a first magnet of the loading station to a position sufficient to exert a first magnetic force on at least some of the magnetic barcode-bearing beads in a first direction, moving a second magnet of the loading station to a position sufficient to exert a second magnetic force on at least some of the magnetic barcode-bearing beads in a second direction different than the first direction
Flush beads out through the outlet port by introducing a fluid
introducing a fluid into the flowcell to cause at least some of the barcode-bearing beads to flow through the flowcell and out of the flowcell through the outlet port
Inferior first magnet maintains beads while lysing cells
moving a first magnet of the loading station from a first magnet inactive position to a position inferior to the flow cell sufficient to exert a first magnetic force on at least some of the magnetic nucleic acid barcode-bearing beads to maintain the at least some of the magnetic nucleic acid barcode-bearing beads positioned within the plurality of microwells, and lysing cells positioned within the plurality of microwells
Superior second magnet releases beads after lysis
moving a second magnet of the loading station from a second magnet inactive position to a position superior to the flow cell to exert a second magnetic force on at least some of the magnetic nucleic acid barcode-bearing beads positioned within the plurality of microwells
Introduce fluid through inlet to move nucleic acid beads to outlet
introducing a fluid into the flowcell through the inlet port to cause at least some of the magnetic nucleic acid barcode-bearing beads to flow through the flowcell and out the outlet port
Across the independent claims, inventive coverage centers on a cartridge flowcell having a fluidic channel and microwells receiving cells and magnetic barcode-bearing beads, use of a loading station with two movable magnets to exert different-direction magnetic forces to control bead positioning, retention, and release, specific positioning of a first magnet inferior to the flowcell for maintenance during lysing, and subsequent bead release and flush through the outlet port by introducing fluid after magnet repositioning and cell lysis.
Stated Advantages
Documented Applications
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