Rotary platform for cell lysing and purification and method of use

Patent metadata

Field	Value
Publication number	US-11638917-B2
Application number	US-202217818144-A
Country	US
Kind code	B2
Filing date	Aug 8, 2022
Priority date	Jul 3, 2019
Publication date	May 2, 2023
Grant date	May 2, 2023

Abstract

Magnetic beads having cell components of interest are translated between a sequence of processing wells in a tray without need for pipetting. The circular tray contains one or more sequences of wells each interconnected by a respective channel. The tray is rotated about a central axis and a magnet, an agitator, and a heater provided external to the tray enable magnetic bead translation, mixing, and incubation, respectively. The magnet proximate a well forms a cluster of beads. Manipulation of the tray in rotation and elevation results in translation of the cluster from one well, through a channel, and into an adjacent well. The well containing a cluster may be rotationally positioned in front of the agitator, the agitator extended into contact with the well, followed by mechanical agitation. The heater, disposed beneath the tray, may accept a well lowered thereto for selective heating.

First claim

We claim:

1. A method of lysing, purifying, and eluting a fluid sample, comprising:
disposing a lysis buffer in a primary well of a sequence of plural wells of a rotary platform on a vertically aligned actuator, the rotary platform comprising
a substantially circular tray having an upper surface, a lower surface, a thickness between the upper surface and the lower surface, an axis of symmetry orthogonal to the upper and lower surfaces, and an outer radial edge,
the sequence of plural wells, each well having an outer surface extending from the lower surface of the tray about a periphery of the tray and having an interior volume and an opening in the tray upper surface, the opening extending into the respective well interior volume, the wells of the sequence of plural wells being disposed in an arc equidistant from the axis of symmetry,
a channel formed in the tray upper surface intermediate each pair of adjacent ones of the plural wells in the sequence and intersecting the opening of each of the plural wells in the respective sequence, the channel having a depth less than the thickness and extending between the openings of the adjacent ones of the plural wells, and the channel describing an arc equidistant from the axis of symmetry, and
a rotor mating interface coaxial with the tray axis of symmetry and in mechanical communication with an actuator,
wherein the channel has inner walls and an outer wall, all points on each of the inner walls having a first identical radial distance from the axis of symmetry and all points on the outer wall having a second radial distance from the axis of symmetry, and each inner wall extends between adjacent wells in the sequence, and
wherein the channel has a floor that is parallel to the tray upper and lower surfaces;

disposing wash buffers in at least one secondary well of the sequence;
disposing an elution buffer in a tertiary well of the sequence;
disposing a fluid sample into the primary well of the sequence;
disposing magnetic beads into the primary well of the sequence;
rotating the tray through selective actuation of the actuator to align the primary well with a magnetic member, which is configured to selectively horizontally extend and retract a magnet with respect to a first position adjacent to the tray;
horizontally extending the magnet, by the magnetic member, adjacent the primary well, whereby the magnetic beads are accumulated into a cluster on an interior wall of the primary well;
lowering the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels up the primary well wall to the respective channel as the tray is lowered;
rotating the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels across the respective channel to the opening of a first of the secondary wells as the tray is rotated;
raising the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels down the well wall of the first of the secondary wells as the tray is raised;
horizontally retracting the magnetic member whereby the cluster of magnetic beads is released;
repeating the sequential steps of extending the magnetic member, lowering the tray, rotating the tray, raising the tray, and retracting the magnet for each subsequent secondary well including a last secondary well;
extending the magnetic member whereby the magnet is adjacent the last secondary well and whereby the magnetic beads are accumulated into a cluster on an interior wall of the last secondary well;
lowering the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels up the last secondary well wall to the respective channel as the tray is lowered;
rotating the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels across the respective channel to the opening of the tertiary well as the tray is rotated;
raising the tray through selective actuation of the actuator, whereby the cluster of magnetic beads, attracted by the magnet, travels down the well wall of the tertiary well as the tray is raised; and
aspirating eluate from the tertiary well.

2. The method of claim 1 , further comprising;
providing an agitating member configured to selectively horizontally extend and retract an agitator with respect to a second position adjacent to the tray; and
rotating the tray through selective actuation of the actuator, whereby the well containing the magnetic beads is aligned with the agitating member;
extending the agitating member whereby the agitator is in mechanical communication with the well containing the magnetic beads;
agitating the well containing the magnetic beads; and
retracting the agitating member.

3. The method of claim 1 , further comprising:
providing a heater at a position radially offset from the axis of symmetry substantially equal to the radial offset of each well in the sequence of wells;
rotating the tray through selective actuation of the actuator to dispose a target well from the plural wells to be heated above the heater;
lowering the tray through selective actuation of the actuator to dispose the target well within the heater;
actuating the heater to heat the target well to a desired temperature or temperature profile for a desired time period; and
raising the tray through selective actuation of the actuator, whereby the target well is retracted from the heater.

4. The method of claim 1 , wherein the outer surface of each well is frustoconical.

5. The method of claim 1 , wherein each well extends substantially orthogonally from the lower surface of the tray.

6. The method of claim 1 , wherein the outer surface of each well narrows in a direction distal from the tray.

7. The method of claim 1 , wherein the opening of each well is substantially circular and coplanar with the upper surface of the tray, and
wherein the outer wall of the channel has a radial distance that is substantially equal to a maximum radial distance, relative to the axis of symmetry, of the opening of each of the adjacent ones of the plural wells in the sequence.

8. The method of claim 7 , wherein each substantially circular opening has a respective center point, and
wherein each inner wall of the channel has a radial distance that is substantially equal to a center-point radial distance, relative to the axis of symmetry, of the center point of each of the adjacent ones of the plural wells in the sequence.

9. The method of claim 1 , wherein the channel has a floor that is parallel to the upper and lower surfaces of the tray.

10. The method of claim 1 , wherein the sequence of plural wells is a first sequence of plural wells and the channel is a first channel, and wherein the rotary platform further comprises a second sequence of plural wells, identical to the first sequence of wells, and a second channel, identical to the first channel.

11. The method of claim 10 , wherein the first and second sequences of plural wells are symmetrically distributed about the periphery of the tray.

12. The method of claim 1 , wherein the secondary wells comprise two secondary wells.

13. A method of lysing, purifying, and eluting a fluid sample, comprising:
disposing a lysis buffer in a primary well of a sequence of plural wells of a rotary platform on a vertically aligned actuator, the rotary platform comprising:
a substantially circular tray having an upper surface, a lower surface, a thickness between the upper surface and the lower surface, an

Assignees

• Siemens Healthcare Diagnostics Inc

Inventors

• Bryant Jason
• Yeaton Eric
• Harris Dan
• Hackendorf Josiah
• Allen Steven
• Talmer Mark

Classifications

• G01N2035/00554

  using ultrasound · CPC title

• B01L3/5085

  for multiple samples, e.g. microtitration plates · CPC title

• G01N35/0098

  involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation (magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C) · CPC title

• B01L3/502 Primary

  with fluid transport, e.g. in multi-compartment structures · CPC title

• B01L2200/0631

  Purification arrangements, e.g. solid phase extraction [SPE] · CPC title