Continuous interleaved process for conducting an assay in a multi-well plate

• US 10,203,286 B2
• Filed: 04/10/2009
• Issued: 02/12/2019
• Est. Priority Date: 04/11/2008
• Status: Active Grant

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Diagnostics, LLC
• Inventors: Clinton, Charles M., Glezer, Eli N., West, Sharon, Sigal, George, Stevens, Carl, Vock, Michael L.
• Primary Examiner(s): Hixson, Christopher Adam
• Assistant Examiner(s): Adams, Michelle
• Application Number:
  US12/422,081
• Publication Number:
  US 20090263904A1
• Time in Patent Office: 3,595 Days
• Field of Search: None
• CPC Class Codes: G01N 2035/00524 Mixing by agitating sample carrier
• CPC Class Codes: G01N 2035/0425 Stacks, magazines or elevators for plates
• CPC Class Codes: G01N 21/69 specially adapted for fluids , e.g. molten metal
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 35/0092 Scheduling
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/1079 with means for piercing stoppers or septums
• CPC Class Codes: Y10T 436/114165 with step of insertion or removal from test line

Abstract

We describe a method for conducting an assay in a multi-well assay plate using an apparatus comprising a light detection subsystem, a liquid handling subsystem, and a plate handling subsystem. This is particularly well suited for conducting automated sampling, sample preparation, and analysis in a multi-well plate assay format. For example, it may be used for automated analysis of particulates in air and/or liquid samples derived therefrom in environmental monitoring.

Claims

1. A method for conducting an assay in a multi-well assay plate using an apparatus comprising (a) a light detection subsystem, (b) a liquid handling subsystem, (c) a plate handling subsystem, and (d) a computer for controlling operation of said apparatus;

said method comprising a computer-implemented continuous interleaved process including the following steps, performed sequentially, for a plurality of individual wells in said multi-well plate;

(i) adding sample to an individual well during a sample addition phase comprising time slice n;

(ii) incubating said sample in said individual well during an incubation phase comprising time slice m;

(iii) adding reagent to said individual well during a reagent addition phase comprising time slice p such that, in said continuous interleaved process, steps (i)-(iii) are carried out on a first well of said plurality of wells, steps (i)-(iii) are repeated on another at least two wells of said plurality of wells after at least step (i) is completed on said first well, and at least step (i) is completed on said another at least two wells of said plurality of wells before step (ii) is completed on said first well;

(iv) tracking, via a software scheduler, status of steps (i)-(iii) for each of said individual wells; and

(v) comparing an assay signal from said individual well to a pre-defined threshold and repeating steps (i)-(iv) if said assay signal differs from said threshold.

2. The method of claim 1, wherein

a first sample is added to said first well in a first sample addition phase;

said first sample is incubated in said first well in a first incubation phase;

during said first incubation phase, a second sample is added to a second well of said plurality of wells in a second sample addition phase;

after said second sample addition phase, reagent is added to said first well in a first reagent addition phase,

during said first reagent addition phase, said second sample in said second well is incubated in a second incubation phase; and

after said first reagent addition phase, reagent is added to said second well during a second reagent addition phase.

3. The method of claim 2, which further comprises automatically repeating steps (i)-(v) on said first sample or said second sample if said first sample or said second sample, respectively, has an assay signal that differs from a pre-defined threshold.

4. The method of claim 3, wherein a sample that has an assay signal that differs from a pre-defined threshold is sent to the front of an assay queue.

5. The method of claim 1, wherein said first well and said another at least two wells of said plurality of wells are contiguous wells of said multi-well plate.

6. The method of claim 1, wherein said first well and said another at least two wells of said plurality of wells are non-contiguous wells of said multi-well plate.

7. The method of claim 1, wherein

step (i) further comprises piercing a seal of said individual well prior to adding sample.

8. The method of claim 1, wherein

step (i) further comprises adding an additional reagent to said individual well.

9. The method of claim 7, wherein

step (i) further comprises adding an additional reagent to said individual well.

10. The method of claim 1, wherein

step (iii) further comprises aspirating fluid from said individual well.

11. The method of claim 1 further comprising step (vi) detecting luminescence from said individual well, and steps (i)-(vi) are carried out on said first well and steps (i)-(vi) are repeated on said another at least two wells of said plurality of wells after at least step (i) is completed on said first well.

12. The method of claim 1 further comprising step (vi) incubating said individual well in a second incubation phase, and (vii) adding a second reagent to said individual well in a second reagent addition phase, wherein steps (i)-(vii) are carried out on said first well and steps (i)-(vii) are repeated on said another at least two wells of said plurality of wells after at least step (i) is completed on said first well.

13. The method of claim 1, which comprises exclusively dedicating said sample addition phase to said time slice n and exclusively dedicating said reagent addition phase to said time slice p.

14. The method of claim 2 further comprising, during a subsequent incubation phase, adding sample to another well of said plurality of wells if sample is available, and idling a pipettor of said apparatus if no sample is available.

15. The method of claim 2 further comprising adding reagent to said individual well during time slice p if said individual well has reached the end of time slice m, and idling said apparatus for time slice p if said individual well is in time slice m and no other individual well has reached the end of its incubation slice m.

16. The method of claim 15, wherein time slices n and p alternate.

17. The method of claim 1, which further comprises random access sampling.

18. The method of claim 1, wherein the steps further include receiving user-configured values for one or more of time slices n, m, or p for one or more wells of said multi-well plate, and via said software scheduler, adjusting a number of said other wells on which said steps (i)-(iii) are repeated.