Assay apparatuses, methods and reagents

• US 10,302,649 B2
• Filed: 12/21/2006
• Issued: 05/28/2019
• Est. Priority Date: 12/21/2005
• Status: Active Grant

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Clinton, Charles M., Glezer, Eli N., Jeffrey-Coker, Bandele, Kovacs, Sandor L., Kumar, Sudeep M., Sigal, George, Stevens, Carl, Vock, Michael
• Primary Examiner(s): Turk, Neil N
• Application Number:
  US11/642,968
• Publication Number:
  US 20070231217A1
• Time in Patent Office: 4,541 Days
• Field of Search: None
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B67B 7/24 Hole-piercing devices
• CPC Class Codes: G01N 2035/0405 manipulating closing or opening means, e.g. stoppers, screw caps, lids or covers
• CPC Class Codes: G01N 2035/0425 Stacks, magazines or elevators for plates
• CPC Class Codes: G01N 2035/1025 Fluid level sensing
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/66 electrically excited, e.g. electroluminescence
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/1002 Reagent dispensers
• CPC Class Codes: G01N 35/1011 Control of the position or alignment of the transfer device
• CPC Class Codes: G01N 35/1079 with means for piercing stoppers or septums
• CPC Class Codes: G01N 35/1081 characterised by the means for relatively moving the transfer device and the containers in an horizontal plane G01N35/1011 takes precedence
• CPC Class Codes: G01N 35/109 with two horizontal degrees of freedom
• CPC Class Codes: Y10T 83/9314 Pointed perforators

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 measuring signals from wells of sealed multi-well assay plates, the apparatus comprising:

(a) a sealed multi-well assay plate comprising wells that are square or square with rounded corners;

(b) a seal removal tool for removing seals from wells of said multi-well plates, wherein said seal removal tool comprises a piercing probe with a seal piercing tip and said piercing probe comprises;

(i) a piercing section with external surfaces that taper to a vertex so as to form said piercing tip at one end of a piercing direction, wherein said piercing section is pyramidal and said piercing section includes exposed cutting edges extending in a radial direction from said piercing tip, wherein said exposed cutting edges are positioned in said piercing section to align with the corners of said wells;

(ii) a seal displacement section, arranged adjacent to said piercing section along said piercing direction, with a cross-sectional shape, perpendicular to said piercing direction, that is square or square with rounded corners, and slightly undersized relative to said wells;

(iii) a plate stop section adjacent to said seal displacement section, wherein said plate stop section is sized to prevent entry of the stop section into said wells, thereby defining the maximal distance of travel of said piercing probe into said wells; and

(iv) a through-hole parallel to said piercing direction and off-set from said piercing tip;

(c) a piercing probe guide having an opening formed therein along which the piercing probe slides in the piercing direction;

(d)(c) a pipetting probe that is fixedly attached to the piercing probe guide and movably located in said through-hole such that it can be withdrawn into said piercing probe during seal removal operations and extended from said piercing probe during pipetting operations; and

(e)(d) a spring disposed in the opening in the piercing probe guide and arranged to couple that couples said pipetting probe to said piercing probe, wherein said spring urges the piercing probe away from the piercing probe guide along the piercing direction and has a spring constant selected such that (i) said pipetting probe does not extend from the through-hole when said piercing probe is fully withdrawn from said wells, (ii) translation of said pipetting probe towards said wells results in cotranslation of said piercing probe and delivery of sufficient force to remove seals on said wells, and (iii) continued translation past the maximal distance of travel of said piercing probe results in compression of the spring and extension of said pipetting probe from the through-hole into said wells.

2. The apparatus of claim 1, wherein said piercing probe is spring loaded such that the maximum downward force, along said piercing direction, of said piercing probe on the plate seal is defined by said spring constant.

3. The apparatus of claim 1, wherein movement of said piercing probe and said pipetting probe are controlled independently.

6. The apparatus of claim 1, wherein said seal displacement section is slightly undersized relative to said wells to allow said piercing section to enter the wells, to cut the seal into sections during piercing, and to fold said sections against each well'"'"'s internal wall.

4. A method of detecting a signal from wells of a multi-well plate using a seal removal tool for removing seals from said wells, a piercing probe guide, a pipetting probe, and a spring;

wherein said wells are square or square with rounded corners, wherein said seal removal tool comprises a piercing probe, and wherein said pipetting probe is coupled to said piercing probe by said spring;

wherein said piercing probe comprises;

(i) a piercing section with external surfaces that taper to a vertex so as to form a piercing tip at one end of a piercing direction, wherein said piercing section is pyramidal and said piercing section includes exposed cutting edges extending in a radial direction from said piercing tip, wherein said exposed cutting edges are positioned in said piercing section to align with the corners of said wells;

(ii) a seal displacement section, arranged adjacent to said piercing section along said piercing direction, with a cross-sectional shape, perpendicular to said piercing direction, that is square or square with rounded corners;

(iii) a plate stop section adjacent to said seal displacement section, wherein said plate stop section is sized to prevent entry of the stop section into said wells, thereby defining the maximal distance of travel of said piercing probe into said wells; and

(iv) a through-hole parallel to said piercing direction and off-set from said piercing tip;

wherein said piercing probe guide has an opening formed therein along which the piercing probe slides in the piercing direction;

wherein said pipetting probe is fixedly attached to the piercing probe guide and movably located in said through-hole such that it can be withdrawn into said piercing probe during seal removal operations and extended from said piercing probe during pipetting operations;

wherein said seal displacement section is slightly undersized relative to said wells to allow said piercing section to enter the well, to cut the seal into sections during piercing, and to fold said sections against each well'"'"'s internal wall; and

wherein said spring is disposed in the opening in the piercing guide probe, said spring urging the piercing probe away from the piercing probe guide along the piercing direction and having has a spring constant selected such that (i) said pipetting probe does not extend from the through-hole when said piercing probe is fully withdrawn from said well, (ii) translation of said pipetting probe towards said well results in cotranslation of said piercing probe and delivery of sufficient force to remove a seal on said well, and (iii) continued translation past the maximal distance of travel of said piercing probe results in compression of the spring and extension of said pipetting probe from the through-hole into said well;

the method comprising;

(a) removing a seal on a well of said multi-well plate by cutting said seal into sections and folding said sections against each well'"'"'s internal wall, wherein said pipetting probe is withdrawn into said piercing probe during said removal step;

(b) extending said pipetting probe from said piercing probe;

(c) pipetting, from said pipetting probe, sample and/or assay reagent into said well;

(d) pipetting, from said pipetting probe, assay reagent into said well; and

(e) detecting said signal from said well.

5. The method of claim 4 further comprising repeating said method on one or more additional wells of said plate.

7. The method of claim 4 further comprising removing a liquid from said well.

8. The method of claim 4 further comprising washing said well.

9. The method of claim 4 further comprising illuminating said well.

10. The method of claim 4 further comprising applying an electrical potential to said well.