Loading station

Patent metadata

Field	Value
Publication number	US-11061043-B2
Application number	US-202016824318-A
Country	US
Kind code	B2
Filing date	Mar 19, 2020
Priority date	Aug 15, 2017
Publication date	Jul 13, 2021
Grant date	Jul 13, 2021

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.

First claim

What is claimed is:

1. A method comprising:
introducing a fluid comprising a plurality of cells into a flowcell of a cartridge positioned within a loading station, the flowcell comprising a fluidic channel, an inlet port, and an outlet port, wherein the fluidic channel comprises a bottom comprising a plurality of microwells, wherein the liquid is introduced into the inlet port of the flowcell and flowed across the plurality of microwells;
introducing a fluid comprising plurality of magnetic barcode-bearing beads into the flowcell, wherein the fluid is introduced into the inlet port of the flowcell and flowed across the plurality of microwells, wherein each microwell is dimensioned to receive at least one cell of the plurality of cells and at least one magnetic barcode-bearing bead of the plurality of magnetic barcode-bearing beads;
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 positioned within the plurality of microwells 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 positioned within the plurality of microwells in a second direction different than the first direction; and
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.

2. The method of claim 1 , wherein at least one of the first magnetic force and the second magnetic force are of a magnitude sufficient to prevent magnetic barcode-bearing beads on which the at least one of the first magnetic force and second magnetic force are exerted from flowing through the flowcell when the fluid is introduced into the flowcell.

3. The method of claim 1 , wherein each of the first magnet and the second magnet are coupled to an actuation mechanism comprising an actuator.

4. The method of claim 3 , wherein moving the first magnet of the loading station to exert a first magnetic force on at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells in a first direction comprises moving the actuator to a first actuator position.

5. The method of claim 4 , wherein moving the second magnet of the loading station to exert a second magnetic force on at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells in a second direction different than the first direction comprises moving the actuator to a second actuator position from the first actuator position.

6. The method of claim 5 , wherein moving the actuator from the first actuator position to the second actuator position causes the first magnet to move to a position in which less magnetic force is exerted on the at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells by the first magnet than when the actuator is positioned within the first position.

7. The method of claim 5 , wherein moving the actuator from the first actuator position to the second actuator position causes the second magnet to move to position in which more magnetic force is exerted on the at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells by the second magnet than when the actuator is positioned within the first position.

8. The method of claim 1 , wherein moving the first magnet of the loading station to the position sufficient to exert the first magnetic force on at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells in the first direction comprises moving the first magnet to a position inferior to the flowcell.

9. The method of claim 1 , wherein moving the second magnet of the loading station to the position sufficient to exert the second magnetic force on at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells in the second direction different than the first direction comprises moving the second magnet to a position superior to the flowcell.

10. The method of claim 1 , wherein the method further comprises lysing cells in the plurality of the microwells between moving of the first and second magnets.

11. The method of claim 1 , wherein the method further comprises positioning the cartridge in the loader.

12. A method comprising:
introducing a fluid comprising a plurality of cells into a flowcell of a cartridge positioned within a loading station, the flowcell comprising a fluidic channel, an inlet port, and an outlet port, wherein the fluidic channel comprises a bottom comprising a plurality of microwells, wherein the liquid is introduced into the inlet port of the flowcell and flowed across the plurality of microwells so that cells are positioned in the microwells;
introducing a fluid comprising a plurality of magnetic nucleic acid barcode-bearing beads into the flowcell, wherein the fluid is introduced into the inlet port of the flowcell and flowed across the plurality of microwells, wherein each microwell is dimensioned to receive at least one cell of the plurality of cells and at least one magnetic nucleic acid barcode-bearing bead of the plurality of magnetic barcode-bearing beads;
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 positioned within the plurality of microwells to maintain the at least some of the magnetic barcode-bearing beads positioned within the plurality of microwells;
lysing cells positioned within the plurality of microwells;
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 microwell; and
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.

13. The method of claim 12 , wherein the method further comprises moving the first magnet to the first magnet inactive position following lysing.

14. The method of claim 12 , wherein the method further comprises moving the second magnet to the second magnet inactive before introducing the 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.

15. The method of claim 12 , wherein the lysing comprises introducing lysing buffer into the inlet port of the flowcell and flowing the lysis buffer across the plurality of microwells.

16. The method of claim 12 , wherein the method further comprises, following lysing cells, introducing a wash fluid into the inlet port of the flow cell and flowing the wash fluid across the plurality of microwells.

17. The method of claim 12 , wherein the method further comprises, introducing a wash fluid into the inlet port of the flow cell and flowing the wash fluid across the plurality of microwells with the second magnet is in the position superior to the flowcell.

18. The method of claim 12 , wherein each of the first magnet and the second magnet are coupled to an actuation mechanism comprising an actuator.

19. The method of claim 12 , wherein moving the firs

Assignees

• Cellular Res Inc

Inventors

• Lai Janice H
• Spuhler Philipp S
• Facer Geoffrey R
• Li Sixing
• Cesar Christopher G

Classifications

• C12N15/1013 Primary

  by using magnetic beads · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• G01N2035/00158

  Elements containing microarrays, i.e. "biochip" · CPC title

• G01N35/1095

  for supplying the samples to flow-through analysers (for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00) · CPC title

• C12N15/1065

  Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags · CPC title