ASSAY APPARATUSES, METHODS AND REAGENTS

• US 20090263904A1
• Filed: 04/10/2009
• Published: 10/22/2009
• Est. Priority Date: 04/11/2008
• Status: Active Grant

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Vock, Michael L., Glezer, Eli N., Clinton, Charles M., Stevens, Carl, West, Sharon, Sigal, George
• Granted Patent: US 10,203,286 B2
• Time in Patent Office: Days
• US Class Current: 436/48
• 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 apparatuses, systems, method, reagents, and kits for conducting assays as well as process for their preparation. They are particularly well suited for conducting automated sampling, sample preparation, and analysis in a multi-well plate assay format. For example, they may be used for automated analysis of particulates in air and/or liquid samples derived therefrom in environmental monitoring.

Claims

1. An apparatus for conducting luminescence assays in multi-well plates, the apparatus comprising (a) a light detection subsystem;

(b) a liquid handling subsystem; and

(c) a plate handling subsystem, wherein said apparatus processes assay samples by a continuous interleaved process.

2. The apparatus of claim 1 further comprising a sample-focused graphical user interface.

3. An apparatus of claim 1, wherein said plate handling subsystem comprises:

(a) a light-tight enclosure comprising;

(i) one or more plate elevators with a plate lifting platform that can be raised and lowered;

(ii) a light-tight enclosure top having one or more plate introduction apertures positioned above said plate elevators and an imaging aperture, wherein said enclosure top comprises a sliding light-tight door for sealing said plate introduction apertures; and

(iii) a plate translation stage for translating a plate in one or more horizontal directions, wherein said stage comprises a plate carriage for supporting the plate, said plate carriage has an opening to allow said plate elevators positioned below the plate carriage to access and lift the plate, and said plate translation stage is configured to position plates below said imaging aperture and to position said plates above said plate elevators;

(b) one or more plate stackers mounted on said enclosure top, above said plate introduction apertures, wherein said plate stackers are configured to receive or deliver plates to said plate elevators; and

(c) a light detector mounted on said enclosure top and coupled to said imaging aperture with a light-tight seal.

4. The apparatus of claim 1, wherein said liquid handling subsystem further comprises a pipetting system for delivering liquids to or removing liquids from the wells of an assay plate in said apparatus.

5. The apparatus of claim 4, wherein

(a) said pipetting system comprises a pipetting probe mounted on a pipette translation stage for translating said pipetting probe in a vertical direction and, optionally, in one or more horizontal directions;

(b) said enclosure top has one or more pipetting apertures;

(c) said sliding light-tight door has one or more pipetting apertures, wherein said sliding light-tight door has a pipetting position where said pipetting apertures in said enclosure top align with said pipetting apertures in said sliding light-tight door; and

(d) said pipette translation stage is mounted on said enclosure top and configured to allow, when said sliding light-tight door is in said pipetting position, lowering said pipetting probe so as to access wells positioned under said pipetting apertures in said enclosure top.

6. The apparatus of claim 5 further comprising a component selected from the group consisting of reagent and/or sample delivery station, reagent and/or sample tube rack, probe wash station, waste station, and combinations thereof, wherein said pipette translation stage is configured to move in one or more horizontal directions to access liquids in and/or deliver liquids to said component.

7. The apparatus of claim 3 further comprising a plate-seal piercing probe.

8. The apparatus of claim 3 further comprising a plate-seal piercing probe, wherein

(i) said enclosure top has a piercing probe aperture;

(ii) said sliding light-tight door has a piercing probe aperture, wherein said sliding light-tight door has a piercing position where said piercing probe aperture in said enclosure top aligns with said piercing probe aperture in said sliding light-tight door; and

(iii) said piercing probe is mounted on said enclosure top and configured to allow, when said sliding light-tight door is in said piercing position, lowering said piercing probe so as to pierce seals on wells positioned under said piercing apertures in said enclosure top.

9. The apparatus of claim 8, wherein said pipette translation stage comprises a probe translation element and said pipette translation stage is configured to travel horizontally to contact said piercing probe with said probe translation element and to travel vertically to lower and raise said piercing probe with said probe translation element.

10. The apparatus of claim 3 further comprising plate contacts for providing electrical energy to electrodes in wells positioned under said light detector.

11. A method for conducting an assay using the apparatus of claim 1, the method comprising:

(a) introducing a multi-well plate to one of said plate stackers,

(b) sliding said sliding light-tight door so as to expose a plate introduction aperture under said one of said plate stackers,

(c) using one of said plate elevators to lower said plate from said one of said plate stackers to said plate carriage,

(d) sliding said sliding light-tight door to seal said plate introduction apertures,

(e) translating said plate carriage to position one or more wells under said light detector,

(f) detecting luminescence from said one or more wells,

(g) sliding said sliding light-tight door to expose at least one of said plate introduction apertures,

(h) translating said plate carriage to position said plate below said one of said plate introduction apertures, and

(i) raising one of said plate elevators to raise said plate to one of said plate stackers;

wherein said apparatus processes a plurality of wells in said multi-well plate in a continuous interleaved process.

12. The method of claim 11 further comprising one or more of pipetting sample and/or reagent into or out of one of said wells, removing seals from one or more of said wells, or applying electrical energy to electrodes in one or more of said wells.

13. The method of claim 11 further comprising the following steps in order:

(a) pipetting sample and/or reagent into a first well of said multi-well plate;

(b) applying electrical energy to electrodes in said first well;

(c) translating said plate carriage to position said first well of said multi-well plate under said light detector;

(d) detecting luminescence from said first well;

and repeating steps (a)-(d) on one or more additional wells of said multi-well plate after step (a) is completed on said first well.

14. The method of claim 11 further comprising the following steps in order:

(a) removing seals from one or more of said wells;

(b) pipetting sample and/or reagent into a first well of said multi-well plate;

(c) applying electrical energy to electrodes in said first well;

(d) translating said plate carriage to position said first well of said multi-well plate under said light detector;

(e) detecting luminescence from said first well;

and repeating steps (a)-(e) on one or more additional wells of said multi-well plate after step (a) is completed on said first well.

15. The method of claim 14, wherein said seal is removed in step (a) from said first well.

16. A method for conducting an assay using the apparatus of claim 1, the method comprising:

(a) introducing a plate to one of said plate stackers,

(b) sliding said sliding light-tight door so as to expose one of said plate introduction apertures,

(c) using one of said plate elevators to lower said plate from said one of said plate stackers to said plate carriage,

(d) sliding said sliding light-tight door to said piercing position,

(e) aligning a well of said plate under said piercing probe and piercing a seal on said well,

(f) sliding said sliding light-tight door to said pipetting position,

(g) using said pipetting probe to introduce and/or remove reagent and/or sample from one or more wells of said plate,

(h) sliding said sliding light-tight door to seal said plate introduction apertures,

(i) translating said plate carriage to position one or more wells under said light detector,

(j) detecting luminescence from said one or more wells,

(k) sliding said sliding light-tight door to expose one of said plate introduction apertures,

(l) translating said plate carriage to position said plate above one of said plate elevators, and

(m) raising said plate elevator to raise said plate to one of said plate stackers;

wherein said apparatus processes a plurality of wells in said multi-well plate in a continuous interleaved process.

17. The method of claim 16 further comprising one or more of pipetting sample and/or reagent into or out of one of said wells, removing seals from one or more of said wells, or applying electrical energy to electrodes in one or more of said wells.

18. The method of claim 16 further comprising the following steps in order:

(a) pipetting sample and/or reagent(s) into a first well of said multi-well plate,

(b) applying electrical energy to electrodes in said first well,

(c) translating said plate carriage to position said first well of said multi-well plate under said light detector, and

(d) detecting luminescence from said first well;

and repeating steps (a)-(d) on one or more additional wells of said multi-well plate after step (a) is completed on said first well

19. The method of claim 16 further comprising the following steps in order:

(a) removing seals from one or more of said wells,

(b) pipetting sample and/or reagent(s) into a first well of said multi-well plate,

(c) applying electrical energy to electrodes in said first well,

(d) translating said plate carriage to position said first well of said multi-well plate under said light detector, and

(e) detecting luminescence from said first well;

and repeating steps (a)-(e) on one or more additional wells of said multi-well plate after step (a) is completed on said first well.

20. The method of claim 19, wherein said seal is removed in step (a) from said first well.

21. The method of claim 10, wherein said light detector is an imaging system.

22. The method of claim 21, wherein said imaging system is used to image luminescence from arrays of binding domains in said one or more wells and said apparatus reports luminescence values for luminescence emitted from individual elements of said arrays.

23. The method of claim 10, wherein one or more wells of said plate comprise dry assay reagents.

24. The method of claim 23, wherein said one or more wells comprising dry assay reagents are sealed to protect said dry reagents from the environment.