Assay devices, methods and reagents

Abstract

Devices, systems, methods, reagents, and kits for performing assays and processes for making them are described. They are particularly suitable for automated analysis in a multi-well plate assay format. A method of focusing an optical sensor to a spaced apart platform is provided, comprising the steps of: providing at least one patterned upper, middle, and lower surface, wherein the patterned middle surface and the platform are aligned with each other, and a first distance between the patterned upper and middle surfaces and a second distance between the middle surface and the patterned lower surface are substantially equal; obtaining a first difference in contrast values between the patterned upper and middle surfaces with the optical sensor; (c) obtaining a second difference in contrast values between the patterned middle and lower surfaces with the optical sensor; and (d) comparing the first and second differences in contrast values.

Description

[0001] This application is a divisional application of patent application No. 201910174980.X, filed on January 3, 2014, entitled "Laboratory Apparatus, Method, and Reagents". Patent application No. 201910174980.X is a divisional application of PCT patent application No. 201480012891.0, filed on January 3, 2014, entitled "Laboratory Apparatus, Method, and Reagents". Cross-references to related applications

[0002] This patent application claims priority to U.S. Provisional Application No. 61 / 749,097, filed January 4, 2013, entitled “Assay Apparatus, Method, and Reagents,” the entire description of which is incorporated herein by reference. Also incorporated herein by reference are U.S. Applications Publication Nos. 2011 / 0143947, 2012 / 0195800, 2007 / 0231217, 2009 / 0263904, and 2011 / 025663, the entire descriptions of which are also incorporated herein by reference. Technical Field

[0003] This invention relates to apparatus, systems, methods, reagents, and complete sets of equipment for performing laboratory tests. Certain embodiments of the apparatus, systems, methods, reagents, and complete sets of equipment of this invention can be used for automated sampling, sample preparation, and / or sample analysis in multi-well plate testing. Background Technology

[0004] Numerous methods and systems have been developed for performing chemical, biochemical, and / or biological assays. These methods and systems are essential in a wide range of applications, including medical diagnostics, dietary testing, environmental monitoring, manufacturing quality control, drug discovery, and fundamental scientific research.

[0005] Multi-well plates (also known as microtiter plates or microplates) have become the standard format for the processing and analysis of multiple samples. Multi-well plates can be available in various forms, sizes, and shapes. For convenience, several standards have been established for instruments used to process large volumes of samples for analysis. Multi-well plates are typically manufactured in standard sizes and shapes and have standard well arrangements. Well arrangements include those found on 96-well plates (12x8 array), 384-well plates (24x16 array), and 1536-well plates (48x32 array). The Society for Biomolecular Screening has published recommended microplate specifications for various plate shapes (see http: / / www.sbsonline.org).

[0006] Various plate reading devices can be used to perform analytical measurements on multi-well plates containing reading devices, measuring changes in light absorption, light emission (e.g., fluorescence, phosphorescence, chemiluminescence, and electrochemiluminescence), radiation, light scattering, and magnetic field. U.S. Patent Application Publication 2004 / 0022677 by Wohlstadter et al. and U.S. Patent No. 7,842,246 describe schemes for single and multiple ECL assays using multi-well plate formats. These include plates comprising a plate top with through-holes forming well walls and a plate bottom sealing the plate top to form well bottoms. The plate bottom has a patterned conductive layer that provides electrode surfaces for the wells, acting as a solid-phase support for the bonding reaction and electrodes for inducing electrochemiluminescence (ECL). The conductive layer may also include electrical contacts for applying electrical energy to the electrode surfaces.

[0007] Despite the existence of these known methods and systems for performing tests, there is still a need for improved devices, systems, methods, reagents, and complete sets of equipment for automated sampling, sample preparation, and / or sample analysis using multi-well plate testing. Summary of the Invention

[0008] Therefore, the present invention provides a method for focusing an optical sensor onto a spaced-out platform, comprising the steps of: (a) providing at least one patterned upper, middle, and lower surface, wherein the patterned middle surface and the platform are aligned with each other, and a first distance between the patterned upper surface and the patterned middle surface and a second distance between the middle surface and the patterned lower surface are substantially equal; (b) obtaining a first difference in contrast values ​​between the patterned upper surface and the middle surface using an optical sensor; (c) obtaining a second difference in contrast values ​​between the patterned middle surface and the lower surface using an optical sensor; and (d) comparing the first difference and the second difference in contrast values.

[0009] Therefore, the present invention also provides a focusing mechanism for an optical sensor, comprising at least one patterned upper, middle, and lower surface spaced apart from the optical sensor; wherein the patterned middle surface is aligned with a target surface to be focused by the optical sensor and the patterned middle surface, wherein a first distance between the patterned upper and middle surfaces is substantially equal to a second distance between the middle and lower surfaces, wherein the optical sensor and the patterned surfaces are moved relative to each other until the difference between a first and a second difference in contrast values ​​between the patterned upper and middle surfaces and between the patterned middle and lower surfaces is less than a predetermined value; and a light source is provided to allow light to pass through the patterned upper, middle, and lower surfaces toward the optical sensor.

[0010] The present invention carefully considers an instrument comprising: (a) a contact platform comprising a plurality of interrogation regions, each interrogation region comprising at least one pair of electrical contacts for applying a potential to the interrogation region; (b) a controller operatively connected to a voltage source, wherein the voltage source is connectable to one or more pairs of electrical contacts; and (c) a multiplexer connected to the controller and the voltage source to selectively connect the voltage source to one pair of electrical contacts in a single interrogation region, or to connect the voltage source to multiple pairs of electrical contacts in more than one interrogation region.

[0011] The instrument of the present invention further includes: (a) a contact platform, wherein the platform includes a plurality of interrogation areas, each interrogation area including at least one pair of electrical contacts conducting an electrical potential to the interrogation area; (b) a controller operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts; and (c) means for connecting the controller and the voltage source to switch from a first connection between the voltage source and the electrical contacts of a single interrogation area to a second connection between the voltage source and the electrical contacts of one or more interrogation areas.

[0012] The instrument is preferably suitable for interrogating samples contained in a multi-well plate and includes: (a) a carriage frame for supporting the multi-well plate and movable relative to a contact platform, wherein the multi-well plate includes a plurality of wells arranged in an MxN matrix, and wherein the contact platform includes a plurality of interrogation regions, wherein each interrogation region includes at least one pair of electrical contacts conducting potential to at least one well; (b) a controller operatively connected to a motor to move the carriage frame relative to the contact platform, and the controller operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts; and (c) a multiplexer connected to the controller and the voltage source to selectively connect the voltage source to a pair of electrical contacts in a single interrogation region, or to at least one pair of electrical contacts in more than one interrogation region.

[0013] Another embodiment of the present invention is a method for querying samples contained in a multi-well plate of wells having an MxN matrix, comprising the steps of: (a) providing a contact platform having a plurality of query regions, (b) providing at least one pair of electrical contacts for each query region, wherein each query region is adapted to query a single well, (c) selectively applying a potential to: (i) one query region to query one or more wells simultaneously, or (ii) multiple query regions to query multiple wells, and (d) moving the multi-well plate relative to the platform to query additional wells.

[0014] In one specific embodiment, the present invention includes an instrument for performing luminescent assays in a multi-well plate. The instrument includes a light detection subsystem and a plate processing subsystem, wherein the plate processing subsystem includes: (a) A light-shielding enclosure, comprising a housing and a movable drawer, wherein (x) The housing includes a housing top, a housing front, one or more plate inlet holes, a detection hole, a sliding light-shielding door for sealing the plate inlet holes, and multiple alignment features, wherein the housing is adapted to receive a movable drawer, and (y) Movable drawers, including: (i) xy subframe, including multiple mating alignment features for engaging and meshing with the numerous alignment features to align a movable drawer within the housing relative to the photodetector subsystem, wherein the weight of the movable drawer is supported by the top of the housing; (ii) One or more plate lifts having a plate lifting platform that can be raised and lowered, wherein the position of the one or more plate lifts is below the plate inlet hole; (iii) A plate translation platform for translating a plate along one or more horizontal directions, wherein the platform includes a plate carriage for supporting the plate, the plate carriage having an opening allowing a plate lifter positioned below the plate carriage to approach and lift the plate, and the plate translation platform is used to position the plate below a detection hole and above the plate lifter; and (b) One or more plate stackers mounted on top of the housing, above the plate inlet holes, wherein the plate stackers are configured to receive plates or transport plates to a plate lifter; and The photodetector subsystem includes a photodetector mounted on the top of the housing and integrated into a detection aperture with a light-shielding seal.

[0015] This instrument can be used for luminescence analysis in multi-well plates and includes a plate handling subsystem. This subsystem includes a plate carriage for supporting the multi-well plate, wherein the plate carriage includes a frame and a plate locking mechanism. The plate locking mechanism includes: (a) Plate carriage flange; (b) A plate clamping arm perpendicular to the flange, including a proximal end and a distal end relative to the flange, wherein the arm is attached to the frame at its proximal end and the arm is rotatable in the XY plane at its distal end, and the arm also includes an upper clamp including an inclined surface configured to engage with the plate. (c) A plate positioning element, comprising a rod, a pedal, and a spring, wherein the rod is substantially perpendicular to the arm, substantially parallel to the flange, and attached to the distal end of the arm via the spring, while the pedal is attached to the rod at an angle; and (d) A plate wall that is substantially parallel to the arm and substantially perpendicular to the positioning element and the flange, and is disposed between the two, the plate wall comprising (i) a lower plate clamp configured to engage with the skirt of the multi-well plate, and (ii) a lower plate clamp ramp for pushing the lower plate clamp toward the skirt.

[0016] The present invention also relates to a method for embedding a multi-well plate into the instrument just discussed above. The method includes the following steps: (a) Place the board on the frame; (b) The spring of the compression plate positioning element pushes the pedal against the flange against the plate and causes the arm to rotate toward the plate in the XY plane; (c) Make the upper clamp contact the plate, thereby pushing the plate against the carriage wall; (d) Make the lower plate clamp contact with the skirt, thereby locking the plate in the carriage.

[0017] Furthermore, the present invention provides an instrument for performing luminescence analysis in a multi-well plate, and includes a plate processing subsystem comprising a plate carriage for supporting the multi-well plate and a plate locking mechanism.

[0018] The multi-well plate has at least one first, second, third, and fourth side surface, with the first and third side surfaces being substantially parallel to each other, and the second and fourth side surfaces being substantially parallel to each other. The plate carriage defines a hole whose shape is basically the same as that of the multi-well plate and whose size is smaller than that of the multi-well plate to support a flange located around the multi-well plate. The plate carriage also includes first (501) and second (513) limiting members corresponding to the first and second sides of the multi-well plate, respectively.

[0019] The plate locking mechanism can be moved from an open configuration for receiving a multi-well plate to a clamping configuration for locking the multi-well plate onto the plate carriage. The plate locking mechanism includes a first locking element (509) biased to a clamping position and a plate clamping arm (502) biased to a clamping position. The first locking element has a pedal (511) adapted to push a first side of the multi-well plate toward a first limiting member. The plate clamping arm has a bracket (503) pivotally connected to the plate clamping arm (502) and adapted to push a second side toward a second limiting member (513). The first locking mechanism (509) is connected to the plate clamping arm (502), and... The plate locking mechanism includes at least one biasing clamp (515) positioned adjacent to the second limiting member (513) to clamp the skirt of the multi-well tray onto the plate carriage.

[0020] Furthermore, the present invention provides a system comprising: (i) Multi-well test plates, selected from the group consisting of single-well addressable plates or multi-well addressable plates; and (ii) A device configured to measure electrochemiluminescence (ECL) from a single well of a single-well addressable plate and a group of wells of a multi-well addressable plate.

[0021] The invention also includes a device for measuring light emission from a multi-well plate, the plate type of which is selected from the group consisting of single-well addressable plates or multi-well addressable plates, the device comprising: (i) Board type identification interface for identifying board type; (ii) A plate translation platform for holding and translating multi-well plates in the XY plane; (iii) A plate contact mechanism comprising a plurality of contact probes and positioned below a plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift that can raise and lower the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) A voltage source that applies a potential to the plate via a contact probe; and (v) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism, wherein (a) The imaging system is configured for PxQ matrix imaging of a well, the plate contact mechanism is configured to contact the bottom contact surface associated with the matrix, and the plate translation stage is configured to translate the plate to position the matrix in alignment with the imaging system and the plate contact mechanism; (b) The device is configured to sequentially apply voltage to each well in a matrix of single-well addressable plates and to perform matrix imaging; and (c) The device is configured to simultaneously apply voltage to each well in a matrix of multi-well addressable plates and to perform matrix imaging.

[0022] A method for measuring the emission from a single-well addressable plate or a multi-well addressable plate is also provided, wherein the method includes: (a) Load the plate onto the plate translation platform; (b) Identify whether the board is a single-well or multi-well addressable board; (c) Move the plate translation stage to align the first PxQ matrix of the well with the plate contact mechanism and the imaging system; (d) A lifting plate contact mechanism so that the contact probe on the contact mechanism contacts the bottom contact surface associated with the PxQ matrix of the well; (e) If the board is a single-well addressable board, then while the group is being imaged, light emission is generated in the PxQ matrix and imaged by sequentially applying voltage to each well in the group; (f) If the board is a multi-well addressable board, then while imaging the matrix, emission is generated in the PxQ matrix and imaged by simultaneously applying voltage to each well in the matrix; and (g) For the additional PxQ matrix in the board, repeat steps (c) to (f).

[0023] In some embodiments, the present invention provides a method for focusing an optical sensor onto spaced-out platforms, comprising the following steps: a. Provide at least one patterned upper, middle and lower surface, wherein the patterned middle surface and the platform are aligned with each other, and a first distance between the patterned upper and middle surfaces is substantially equal to a second distance between the middle surface and the patterned lower surface; b. Obtain the first contrast difference between the patterned upper and middle surfaces using an optical sensor; c. Obtain the second contrast difference between the patterned middle and lower surfaces using an optical sensor; d. Compare the difference between the first and second contrast values.

[0024] In some embodiments, the method further includes step (e) adjusting the distance between the optical sensor and the platform, and repeating steps (b) to (d) until the difference between the first and second contrast values ​​is less than a predetermined value.

[0025] In a method according to some embodiments, the patterned middle surface is substantially aligned with a plane at the same level as the bottom surface of a tray that carries at least one sample to be interrogated by an optical sensor.

[0026] In a method according to some embodiments, the platform includes a plurality of electrodes that contact the bottom surface of a tray to conduct current to at least one sample.

[0027] In the method according to some embodiments, the patterned upper, middle and lower surfaces are located on parallel planes.

[0028] In the method according to some embodiments, the patterned upper, middle, and lower surfaces comprise substantially the same pattern.

[0029] In some embodiments of the method, the same pattern includes a grid.

[0030] In the method according to some embodiments, the predetermined value is less than about ±4.0.

[0031] In the method according to some embodiments, the predetermined value is less than about ±3.0.

[0032] In the method according to some embodiments, the predetermined value is less than about ±2.0.

[0033] In the method according to some embodiments, the distance between the patterned intermediate surface and the platform is between about 4 mm and about 4.75 mm.

[0034] In the method according to some embodiments, the distance is between about 4.5 mm and about 4.7 mm.

[0035] In the method according to some embodiments, the distance is between about 4.6 mm and about 4.7 mm.

[0036] In the method according to some embodiments, the patterned upper surface, middle surface and lower surface are positioned adjacent to each other.

[0037] In the method according to some embodiments, the patterned upper, middle and lower surfaces are located in one quadrant.

[0038] In the method according to some embodiments, the patterned upper, middle, and lower surfaces are illuminated by a light source positioned opposite the optical sensor.

[0039] In the methods according to some embodiments, the optical sensor includes a camera, a CCD sensor, or a CMOS sensor.

[0040] In some embodiments, the present invention provides a focusing mechanism for an optical sensor, comprising at least a patterned upper surface, a middle surface, and a lower surface spaced apart from the optical sensor; The patterned mid-surface is aligned with the target surface that needs to be focused by the optical sensor and the patterned mid-surface. The first distance between the patterned upper and middle surfaces is substantially equal to the second distance between the middle surface and the patterned lower surface. The optical sensor and the patterned surface move relative to each other until the difference between the first and second contrast values ​​is less than a predetermined value; and The light source is positioned such that light passes through patterned upper, middle, and lower surfaces and is directed toward the optical sensor.

[0041] In a focusing mechanism according to some embodiments, the target surface includes a reference plane of the platform, which selectively conducts current to the sample being interrogated by the optical sensor.

[0042] In a focusing mechanism according to some embodiments, the patterned middle surface is aligned with the reference surface of the platform by a predetermined amount.

[0043] In a focusing mechanism according to some embodiments, the distance is between about 4 mm and about 4.75 mm.

[0044] In a focusing mechanism according to some embodiments, the distance is between about 4.5 mm and about 4.7 mm.

[0045] In a focusing mechanism according to some embodiments, the distance is between about 4.6 mm and about 4.7 mm.

[0046] In a focusing mechanism according to some embodiments, the target surface includes the bottom surface of a tray that holds at least one sample to be interrogated by an optical sensor.

[0047] In a focusing mechanism according to some embodiments, the patterned middle surface and the bottom surface of the tray are substantially aligned with the same horizontal plane.

[0048] In a focusing mechanism according to some embodiments, the patterned upper, middle, and lower surfaces are positioned close to each other.

[0049] In a focusing mechanism according to some embodiments, the patterned upper, middle, and lower surfaces are located in one quadrant.

[0050] In a focusing mechanism according to some embodiments, the optical sensor includes a camera, a CCD sensor, or a CMOS sensor.

[0051] In a focusing mechanism according to some embodiments, the patterned upper, middle, and lower surfaces are located on parallel planes.

[0052] In a focusing mechanism according to some embodiments, the patterned upper, middle, and lower surfaces comprise substantially the same pattern.

[0053] In a focusing mechanism according to some embodiments, the same pattern includes a grid.

[0054] According to some embodiments, the present invention provides an instrument comprising: A contact platform, wherein the contact platform includes multiple interrogation areas, and each interrogation area includes at least one pair of electrical contacts that conduct potential to the interrogation area, and A controller that is operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts, and A multiplexer connected to a controller and a voltage source to selectively connect the voltage source to a pair of electrical contacts in a single interrogation area, or to multiple pairs of electrical contacts in more than one interrogation area.

[0055] In an instrument according to some embodiments, multiple interrogation regions are arranged in a PxQ matrix.

[0056] In the instrument according to some embodiments, the PxQ matrix is ​​a 2x2 matrix.

[0057] In the instrument according to some embodiments, multiple pairs of electrical contacts on the contact platform include upright pins.

[0058] In some embodiments of the instrument, the upright pin is spring-loaded.

[0059] The instrument according to some embodiments also includes an optical sensor located above the contact platform.

[0060] In some embodiments of the instrument, a first alignment mechanism is also included, which includes a light source projected from the platform toward an optical sensor to align the platform relative to the optical sensor.

[0061] In some embodiments of the instrument, a plate carriage platform is also included, which is adapted to transport multiple well plates and position the plates relative to a contact platform so that an electric potential can be applied to one or more wells on the plate.

[0062] In some embodiments of the instrument, a second alignment mechanism is also included, which includes a plurality of holes located on the plate carriage frame, and wherein a light source shines from the platform through the holes to further align the plate carriage frame with the contact platform.

[0063] In the instrument according to some embodiments, the plate carriage frame includes a rectangular opening whose specifications and dimensions are made suitable for supporting the skirt around the multi-well plate.

[0064] In an instrument according to some embodiments, the plurality of holes are positioned on at least two sides of a rectangular opening.

[0065] In the instrument according to some embodiments, the plate carriage frame includes a locking mechanism to hold multiple well plates to the plate carriage frame.

[0066] In some embodiments of the instrument, a focusing mechanism located on the plate carriage frame is also included, which allows the optical sensor to be focused relative to the focusing mechanism.

[0067] In some embodiments of the instrument, a third alignment mechanism is also included, which includes a conductive surface located on the plate carriage frame, such that when an electrical contact on the contact platform comes into contact with the conductive surface, current flows between the electrical contacts on the contact platform to indicate a predetermined distance between the electrical contacts and the plate carriage frame.

[0068] In some embodiments, the present invention further provides an instrument comprising: A contact platform, wherein the platform includes multiple interrogation areas, and each interrogation area includes at least one pair of electrical contacts for conducting electrical potential to the interrogation area, and A controller operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts, and A device connected to a controller and a voltage source to switch from a first connection between the voltage source and electrical contacts of a single interrogation area to a second connection between the voltage source and electrical contacts of one or more interrogation areas.

[0069] In some embodiments, the present invention provides an instrument suitable for querying samples contained in a multi-well plate, comprising: The carriage frame is configured to support multiple well plates, and this carriage frame is movable relative to the contact platform. The multi-well plate comprises multiple wells arranged in an MxN matrix. The contact platform includes multiple interrogation areas, each interrogation area including at least one pair of electrical contacts that conduct potential to at least one well. A controller, operatively connected to a motor to move the carriage frame relative to the contact platform, and operatively connected to a voltage source capable of connecting to one or more pairs of electrical contacts, and A multiplexer connected to the controller and the voltage source is configured to selectively connect the voltage source to the pair of electrical contacts in a single interrogation area, or to connect the voltage source to at least one pair of electrical contacts in more than one interrogation area.

[0070] In an instrument according to some embodiments, the interrogation regions are arranged as a PxQ matrix, and the MxN matrix is ​​larger than the PxQ matrix.

[0071] In the instrument according to some embodiments, the PxQ matrix is ​​a 2x2 matrix.

[0072] In an instrument according to some embodiments, a multi-well plate includes bottom electrical contacts for each well located on the bottom surface of the plate, wherein the bottom electrical contacts are adapted to contact multiple pairs of electrical contacts on a contact platform.

[0073] In some embodiments of the instrument, the multi-well plate also includes internal electrical contacts connected to bottom electrical contacts to conduct electrical potential into the well.

[0074] In the instrument according to some embodiments, the bottom electrical contact for at least one well is electrically insulated from the bottom electrical contact of an adjacent well.

[0075] In the instrument according to some embodiments, the internal electrical contacts for at least one well are electrically insulated from the bottom electrical contacts of adjacent wells.

[0076] In the instrument according to some embodiments, the electrical contacts on the platform include a plurality of working electrodes that are selectively connected by a controller to a voltage source to determine the number of wells to be queried.

[0077] In the instrument according to some embodiments, one of the working electrodes is connected to a well.

[0078] In an instrument according to some embodiments, one of the working electrodes is connected to multiple wells.

[0079] In the instrument according to some embodiments, unconnected working terminals are electrically insulated in the multiplexer.

[0080] In the instrument according to some embodiments, the electrical contacts on the platform also include a plurality of counter electrodes electrically connected to at least one ground wire.

[0081] In the instrument according to some embodiments, the bottom electrical contacts of the multi-well tray for multiple wells are electrically connected to the counter electrodes on the contact platform.

[0082] In the instrument according to some embodiments, the bottom electrical contacts of the multi-well tray for all wells are electrically connected to the counter electrodes on the contact platform.

[0083] In the instrument according to some embodiments, the bottom electrical contacts of the multi-well tray for at least one well, which are connected to the counter electrode on the contact platform, are electrically insulated.

[0084] In the instrument according to some embodiments, multiple pairs of electrical contacts on the contact platform include upright pins.

[0085] In some embodiments of the instrument, the upright pin is spring-loaded.

[0086] In the instrument according to some embodiments, the controller simultaneously queries the number of wells of PxQ.

[0087] In instruments according to some embodiments, the controller simultaneously queries fewer than the number of wells (PxQ).

[0088] In some embodiments, the present invention provides a method for querying samples contained in a multi-well plate of a well having an MxN matrix, comprising the following steps: (a) Provide a contact platform with multiple inquiry areas, (b) Provide at least one pair of electrical contacts for each interrogation area, wherein each interrogation area is suitable for interrogating a single well. (c) Selectively apply a potential to: (i) a query region to simultaneously query one or more wells, or (ii) multiple query regions to query multiple wells, and (d) Move the multi-well plate relative to the platform to query other wells.

[0089] In a method according to some embodiments, the query regions are arranged into a PxQ matrix, and the MxN matrix is ​​larger than the PxQ matrix.

[0090] In the method according to some embodiments, in step (c), (i) a single well is queried.

[0091] In the method according to some embodiments, in step (c), (ii) the number of wells of number MxN is queried.

[0092] In some embodiments of the method, step (e) is further included, which involves connecting at least one positive terminal of a plurality of pairs of electrical contacts on a contact platform to a potential, thereby controlling the potential applied in step (c).

[0093] In the method according to some embodiments, step (e) further includes the step of electrically insulating at least one positive terminal not connected to a potential.

[0094] In some embodiments of the method, step (f) is further included in providing bottom electrical contacts on the bottom surface of the multi-well plate.

[0095] In some embodiments of the method, step (g) is further included to electrically insulate at least one ground from the bottom electrical contact.

[0096] In the method according to some embodiments, in step (g), all grounding and bottom electrical contacts are electrically insulated from each other.

[0097] In some embodiments of the method, step (h) is further included: providing an optical sensor.

[0098] In some embodiments of the method, step (i) is further included: focusing the optical sensor onto the contact platform.

[0099] In some embodiments of the method, step (j) is further included to align the optical sensor with the contact platform.

[0100] In some embodiments of the method, step (k) is also included to align a pallet carriage suitable for carrying a multi-well pallet with a contact platform.

[0101] In some embodiments of the method, step (1) is further included to align multiple pairs of electrical contacts of the contact platform with the bottom surface of the multi-well tray.

[0102] In the method according to some embodiments, step (1) further includes providing a conductive surface of the tray carriage frame.

[0103] In some embodiments of the method, step (m) is also included: locking the multi-well tray to the tray carriage frame.

[0104] In some embodiments, the present invention provides an instrument for luminescent assays in a multi-well plate, the instrument comprising a light detection subsystem and a plate processing subsystem, wherein the plate processing subsystem includes: (a) A light-shielding enclosure, comprising a housing and a movable drawer, wherein (x) The housing includes a housing top, a housing front, one or more plate inlet holes, a detection hole, a sliding light-shielding door for sealing the plate inlet holes, and multiple alignment features, wherein the housing is adapted to receive a movable drawer, and (y) Movable drawers, including: (i) an xy subframe comprising a plurality of mating alignment features configured to engage and mesh with the plurality of alignment features to align a movable drawer within the housing relative to the photodetector subsystem, wherein the weight of the movable drawer is supported by the top of the housing; (ii) One or more plate lifts having a plate lifting platform that can be raised and lowered, wherein the position of the one or more plate lifts is below the plate inlet hole; (iii) A plate translation platform for translating a plate along one or more horizontal directions, wherein the platform includes a plate carriage for supporting the plate, the plate carriage having an opening allowing a plate lifter positioned below the plate carriage to approach and lift the plate, and the plate translation platform is configured to position the plate below a detection hole and above the plate lifter; and (b) One or more plate stackers mounted on top of the housing, above the plate inlet holes, wherein the plate stackers are configured to receive plates or transport plates to a plate lifter; and The photodetector subsystem includes a photodetector mounted on the top of the housing and integrated into a detection aperture with a light-shielding seal.

[0105] In some embodiments of the instrument, the xy subframe is mounted on top of the housing.

[0106] In an instrument according to some embodiments, the plurality of alignment features include a set of alignment pins that are always distributed within the housing, and the plurality of mating alignment features include a set of holes configured to engage with the set of alignment pins.

[0107] In an instrument according to some embodiments, the housing includes a plurality of electrical connectors, and the drawer includes a plurality of mating electrical connectors configured to contact and engage with the plurality of electrical connectors.

[0108] In the apparatus according to some embodiments, one or more plate stackers are mobile.

[0109] The instrument according to some embodiments also includes a plate stacker extension element configured to accommodate a plurality of plates, wherein the plate stacker extension element is placed on top of the plate stacker.

[0110] In some embodiments of the instrument, the height of the plate stacker extension element is adjustable.

[0111] In some embodiments of the instrument, the plate stacker extension element is configured to accommodate up to 20 plates.

[0112] In some embodiments of the instrument, the plate stacker extension element is configured to accommodate up to 10 plates.

[0113] In some embodiments of the instrument, the plate stacker extension element is configured to accommodate up to 5 plates.

[0114] In some embodiments of the instrument, the board processing subsystem further includes a board sensor configured to detect boards in the subsystem.

[0115] In instruments according to some embodiments, the plate sensor includes a capacitive sensor, a contact switch, an ultrasonic sensor, a weight sensor, or an optical sensor.

[0116] In the instrument according to some embodiments, the drawer also includes a plurality of overflow collection mechanisms adjacent to one or more plate lifts and / or plate translation platforms.

[0117] In some embodiments of the instrument, the overflow collection mechanism includes a drip protection device.

[0118] In the instruments according to some embodiments, one or more plate lifting platforms include a non-slip surface.

[0119] In some embodiments of the instrument, one or more plate lifts include two adjacent lifting platforms connected by a cross mechanism for raising and lowering the lifting platforms.

[0120] In the instrument according to some embodiments, the plate translation stage includes a locking mechanism configured to prevent movement of the stage.

[0121] In the instrument according to some embodiments, the plate processing subsystem further includes one or more plate orientation sensors.

[0122] In the instrument according to some embodiments, one or more plate orientation sensors are placed on or connected to a plate carriage, the one or more plate stackers and combinations thereof.

[0123] In the instrument according to some embodiments, the light detection subsystem includes a camera and a camera focusing mechanism for focusing the camera in the x, y, z, and θ directions.

[0124] In some embodiments of the instrument, the camera focusing mechanism includes manually adjustable elements.

[0125] In some embodiments of the instrument, manually adjustable elements include knobs.

[0126] In some embodiments of the instrument, the light detection subsystem further includes a motor configured to drive a camera focusing mechanism.

[0127] In the instrument according to some embodiments, the photodetector subsystem includes a photodetector subsystem housing mounted to the board processing subsystem.

[0128] In some embodiments of the instrument, the light detection subsystem housing includes a clip that is fixed to the top of the light-shielding outer housing.

[0129] In some embodiments of the instrument, the clip also includes one or more light-shielding materials configured to prevent light from leaking from the light-shielding housing.

[0130] In some embodiments of the instrument, the housing of the light detection subsystem also includes a light shield surrounding the camera.

[0131] In some embodiments of the instrument, the camera includes a lens with an F-number in the range of 1.3-1.8.

[0132] In the instrument according to some embodiments, the housing includes an internal cooling fan, an air inlet located on a first side of the housing, and an exhaust port located on the opposite side of the housing relative to the first side.

[0133] In some embodiments, the present invention provides an instrument for performing luminescence assays on a multi-well plate, the instrument including a plate processing subsystem, the plate processing subsystem including a plate carriage for supporting the multi-well plate, wherein the plate carriage includes a frame and a plate locking mechanism, the plate locking mechanism including: (a) Plate carriage flange; (b) A plate clamping arm perpendicular to the flange, including a proximal end and a distal end relative to the flange, wherein the proximal end of the arm is attached to the frame and the distal end of the arm is rotatable on the XY plate, and the arm also includes an upper clamp including an inclined surface configured to engage with the plate. (c) A plate positioning element, comprising a rod, a pedal, and a spring, wherein the rod is perpendicular to the arm, parallel to the flange, and attached to the distal end of the arm by means of the spring, while the pedal is attached to the rod at an angle; and (d) A plate wall parallel to the arm, perpendicular to the positioning element and the flange, and disposed between the two, the plate wall including (i) a lower plate clamp for engaging with the skirt of the multi-well plate, and (ii) a lower plate clamp ramp configured to push the lower plate clamp toward the skirt.

[0134] In the instrument according to some embodiments, the plate wall also includes a plate discharge element configured to disengage the lower plate clamp from the skirt.

[0135] In some embodiments, the present invention provides a method for embedding a multi-well plate in an instrument according to some embodiments, comprising: (a) Place the board on the frame; (b) The spring of the compression plate positioning element pushes the pedal against the plate toward the flange and causes the arm to rotate toward the plate in the XY plane; (c) Make the upper clamp contact the plate, thereby pushing the plate against the carriage wall; (d) Make the lower plate clamp contact with the skirt, thereby locking the plate in the carriage.

[0136] In some embodiments, the present invention provides an instrument for performing luminescence analysis on a multi-well plate, the instrument comprising a plate processing subsystem and a plate locking mechanism, the plate processing subsystem comprising a plate carriage supporting the multi-well plate. The multi-well plate has at least a first, second, third, and fourth side surface, with the first and third side surfaces being substantially parallel to each other, and the second and fourth side surfaces being substantially parallel to each other. The plate carriage defines a hole whose shape is substantially the same as that of the multi-well plate, but whose size is smaller than that of the multi-well plate to support the flange located around the multi-well plate. The plate carriage also includes first (501) and second (513) limiting members corresponding to the first and second sides of the multi-well plate, respectively. The plate locking mechanism can be moved from an open configuration for receiving a multi-well plate to a clamping configuration for locking the multi-well plate to the plate carriage. The plate locking mechanism includes a first locking element (509) biased to a clamping position and a plate clamping arm (502) biased to a clamping position. The first locking element has a pedal (511) adapted to push a first side of the multi-well plate toward a first stop. The plate clamping arm has a bracket (503) pivotally connected to the plate clamping arm (502) and adapted to push a second side toward a second stop (513). The first locking mechanism (509) is connected to the plate clamping arm (502), and... The plate locking mechanism includes at least one bias clamp (515) positioned adjacent to the second limiter (513) to clamp the skirt of the multi-well tray to the plate carriage.

[0137] In the instrument according to some embodiments, the bracket (503) includes at least two legs (504, 506), both of which contact the fourth side of the multi-well tray.

[0138] In the instrument according to some embodiments, at least one leg (504, 506) includes a ramp (507, 508) to apply force perpendicular to the plane of the multi-well tray.

[0139] In the instrument according to some embodiments, the first locking element includes an actuating rod (510) biased to a clamping position by a spring (512).

[0140] In the instrument according to some embodiments, a pedal (511) is attached to an actuating rod (510), and the plate carriage has a third limiting member to push the pedal toward and away from the multi-well tray when the actuating rod is moved.

[0141] In the instrument according to some embodiments, the pedal and plate clamp arm are retracted into an opening configuration.

[0142] In some embodiments of the instrument, a discharge device is also included that moves the multi-well tray away from the first or second limiting member.

[0143] In some embodiments of the instrument, the discharge device is pressurized by a spring.

[0144] In some embodiments of the instrument, the discharger includes an over-drift protector.

[0145] In some embodiments, the present invention provides a system comprising: (i) Multi-well test plates, selected from the group consisting of single-well addressable plates or multi-well addressable plates; and (ii) A device configured to measure electrochemiluminescence (ECL) from a single well of a single-well addressable plate and a group of wells of a multi-well addressable plate.

[0146] In a system according to some embodiments, a multi-well assay plate includes a single-well addressable plate comprising a top plate having a top hole and a bottom plate cooperating with the top plate to define wells of the single-well addressable plate. The bottom plate includes a substrate having a top surface and a bottom surface, the top surface having patterned electrodes and the bottom surface having patterned electrical contacts, wherein the electrodes and contacts form a pattern to define a plurality of well bottoms of the single-well addressable plate, wherein the pattern within the well bottoms includes: (a) A working electrode on the top surface of the substrate, wherein the working electrode is electrically connected to an electrical contact; and (b) A counter electrode on the top surface of the substrate, wherein the counter electrode is electrically connected to an electrical contact rather than to other counter electrodes in other wells of a single-well addressable board.

[0147] In systems according to some embodiments, the single-well addressable board is a 4-well board, a 6-well board, a 24-well board, a 96-well board, a 384-well board, a 1536-well board, a 6144-well board, or a 9600-well board.

[0148] In systems according to some embodiments, electrodes and contacts can be addressed independently.

[0149] In some embodiments of the system, the electrodes comprise carbon particles.

[0150] In some embodiments of the system, the electrodes comprise printed conductive material.

[0151] In a system according to some embodiments, one or more electrodes include a plurality of test zones formed thereon.

[0152] In a system according to some embodiments, the plurality of testing areas includes at least four testing areas.

[0153] In a system according to some embodiments, the plurality of testing areas includes at least seven testing areas.

[0154] In a system according to some embodiments, the plurality of testing areas includes at least 10 testing areas.

[0155] In a system according to some embodiments, the plurality of test areas are defined by openings in one or more dielectric layers supported on a working electrode.

[0156] In a system according to some embodiments, a multi-well test plate includes a multi-well addressable plate comprising a top plate having a top hole and a bottom plate cooperating with the top plate to define wells of the multi-well addressable plate. The bottom plate includes a substrate having a top surface and a bottom surface, the top surface having patterned electrodes and the bottom surface having patterned electrical contacts, wherein the electrodes and contacts are patterned to define two or more independently addressable portions of two or more jointly addressable test wells, each portion including two or more wells and: (a) Jointly addressable working electrodes located on the top surface of the substrate, wherein each working electrode is electrically connected to each other and connected to at least the first of the electrical contacts; and (b) Jointly addressable counter electrodes located on the top surface of the substrate, wherein each of the counter electrodes is electrically connected to each other but not to the working electrode, and is connected to at least the second of the electrical contacts.

[0157] In systems according to some embodiments, the multi-well addressable board is a 4-well board, a 6-well board, a 24-well board, a 96-well board, a 384-well board, a 1536-well board, a 6144-well board, or a 9600-well board.

[0158] In a system according to some embodiments, the independently addressable portion includes less than 50% of the wells of a multi-well addressable plate.

[0159] In a system according to some embodiments, the independently addressable portion includes less than 20% of the wells of a multi-well addressable plate.

[0160] In a system according to some embodiments, the independently addressable portion includes a well of a 4x4 array.

[0161] In a system according to some embodiments, the multi-well plate includes independently addressable portions of a 2x3 array.

[0162] In a system according to some embodiments, the independently addressable portion includes one or more rows or one or more columns of wells.

[0163] In some embodiments of the system, the electrodes comprise carbon particles.

[0164] In some embodiments of the system, the electrodes comprise printed conductive material.

[0165] In a system according to some embodiments, one or more electrodes include a plurality of test zones formed thereon.

[0166] In a system according to some embodiments, the plurality of testing areas includes at least four testing areas.

[0167] In a system according to some embodiments, the plurality of testing areas includes at least seven testing areas.

[0168] In a system according to some embodiments, the plurality of testing areas includes at least 10 testing areas.

[0169] In a system according to some embodiments, the device includes: A contact platform, wherein the contact platform includes multiple interrogation areas, and each interrogation area includes at least one pair of electrical contacts for conducting potential to the interrogation area, and A controller that is operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts, and A multiplexer connected to a controller and a voltage source allows for the selective connection of a voltage source to a single pair of electrical contacts in a single interrogation area, or to multiple pairs of electrical contacts in more than one interrogation area.

[0170] In a system according to some embodiments, multiple query regions are arranged as a PxQ matrix.

[0171] In a system according to some embodiments, the PxQ matrix is ​​a 2x2 matrix.

[0172] In a system according to some embodiments, multiple pairs of electrical contacts on the contact platform include upright pins.

[0173] In some embodiments of the system, the upright pin is spring-loaded.

[0174] In some embodiments of the system, an optical sensor located above the contact platform is also included.

[0175] In some embodiments of the system, a first alignment mechanism is also included, which includes a light source that directs light from the contact platform toward the optical sensor to align the contact platform with the optical sensor.

[0176] In systems according to some embodiments, a plate carriage platform is also included, which is adapted to transport and position the multi-well plate relative to a contact platform so that an electric potential can be applied to one or more wells on the multi-well plate.

[0177] In some embodiments of the system, a second alignment mechanism is also included, which includes a plurality of holes located on the plate carriage frame, through which a light source shines from the contact platform to further align the plate carriage frame with the platform.

[0178] In systems according to some embodiments, the plate carriage frame includes a rectangular opening whose specifications and dimensions are made to support the skirt around the multi-well plate.

[0179] In a system according to some embodiments, a plurality of holes are positioned on at least two sides of a rectangular opening.

[0180] In systems according to some embodiments, the plate carriage includes a locking mechanism to hold multiple well plates to the plate carriage frame.

[0181] In some embodiments of the system, a focusing mechanism located on the plate carriage frame is also included to allow the optical sensor to be focused relative to the focusing mechanism.

[0182] In some embodiments of the system, a third alignment mechanism is also included, which includes a conductive surface located on the plate carriage frame such that when an electrical contact on the contact platform comes into contact with the conductive surface, current flows between the electrical contacts on the contact platform to indicate a predetermined distance between the electrical contacts and the plate carriage frame.

[0183] In a system according to some embodiments, the device includes: The platform includes multiple interrogation areas, and each interrogation area includes at least one pair of electrical contacts for conducting potential to the interrogation area. A controller that is operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts, and A device connected to a controller and a voltage source to switch from a first connection between the voltage source and electrical contacts of a single interrogation area to a second connection between the voltage source and electrical contacts of one or more interrogation areas.

[0184] In some embodiments, the present invention discloses an apparatus for measuring light emission from a multi-well plate, wherein the plate type of the multi-well plate is selected from a group consisting of single-well addressable plates or multi-well addressable plates, and the apparatus includes: (i) Board type identification interface for identifying board type; (ii) A plate translation platform for holding and translating a multi-well plate in the xy plane; (iii) A plate contact mechanism including multiple contact probes, the plate contact mechanism being positioned below the plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift capable of raising and lowering the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) A voltage source that applies a potential to the plate via a contact probe; and (v) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism, wherein: (a) The imaging system is configured to image the PxQ matrix of the well, the plate contact mechanism is configured to contact the bottom contact surface associated with the matrix, and the plate translation stage is configured to translate the plate to a position where the matrix is ​​aligned with the imaging system and the plate contact mechanism. (b) The device is configured to sequentially apply voltage to each well in a matrix of single-well addressable plates and to perform matrix imaging; and (c) The device is configured to simultaneously apply voltage to each well in a matrix of multi-well addressable plates and to image the matrix.

[0185] In the apparatus according to some embodiments, the PxQ matrix is ​​a well of a 2x2 array.

[0186] In an apparatus according to some embodiments, the imaging system collects separate images of the voltage of each well in a matrix of continuously applied plates to a single well addressable plate.

[0187] In some embodiments of the apparatus, the board type identification interface includes devices selected from the group consisting of a barcode reader, an EPROM reader, an EEPROM reader, or an RFID reader.

[0188] In the apparatus according to some embodiments, the board type identification interface includes a graphical user interface configured to allow a user to input board type identification information.

[0189] In some embodiments, the present invention provides a method for measuring light emission from a multi-well plate, wherein the plate type of the multi-well plate is selected from a group consisting of single-well addressable plates or multi-well addressable plates, and the apparatus includes: (i) Board type identification interface for identifying board type; (ii) A plate translation platform for holding and translating a multi-well plate in the xy plane; (iii) A plate contact mechanism including multiple contact probes, the plate contact mechanism being positioned below the plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift capable of raising and lowering the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) A voltage source that applies a potential to the plate via a contact probe; and (iv) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism. The method includes: (a) Load the plate onto the plate translation platform; (b) Identify whether the board is a single-well or multi-well addressable board; (c) Move the plate translation stage to align the first PxQ matrix of the well with the plate contact mechanism and the imaging system; (d) A lifting plate contact mechanism so that the contact probe on the contact mechanism contacts the bottom contact surface associated with the PxQ matrix of the well; (e) If the plate is a single-well addressable plate, then while the group is being imaged, light emission is generated in the PxQ matrix and imaged by sequentially applying voltage to each well in the group; (f) If the board is a multi-well addressable board, then while the matrix is ​​being imaged, light emission is generated and imaged in the PxQ matrix by simultaneously applying voltage to each well in the matrix; and (g) For the other PxQ matrices in the board, repeat steps (c) through (f).

[0190] In the method according to some embodiments, the PxQ matrix is ​​a 2x2 array of wells.

[0191] In a method according to some embodiments, the imaging system collects separate images of the voltage of each well in a matrix of continuously applied plates to a single well addressable plate. Attached Figure Description

[0192] Figures 1(a)-(b) show the front and rear views of the device 100 with a lid of a specific style, respectively, and Figures 1(c)-(d) show the corresponding front and rear views of the device without a lid, respectively.

[0193] Figures 2(a)-(c) show detailed diagrams of the board processing subsystem and the photodetector subsystem.

[0194] Figure 3 A diagram showing the movable drawer of the board handling subsystem within the device 100.

[0195] Figures 4(a)-(f) show detailed views of the movable drawer 240 and the various sub-components located inside the drawer.

[0196] Figures 5(a)-(o) show detailed diagrams of the plate carriage and the plate locking mechanism.

[0197] Figures 6(a)-(b) show two alternative embodiments of the optical focusing mechanism that can be incorporated into the device.

[0198] Figures 7(a)-(l) show detailed diagrams of the plate contact mechanism.

[0199] Figures 8(a)-(c) show the various components of the optical detection subsystem.

[0200] Figure 9 This illustrates a non-limiting embodiment of a lens structure that can be used in a photodetector subsystem. Detailed Implementation

[0201] The detailed description section describes certain embodiments of the invention, but this should not be considered limiting, but rather used to illustrate certain aspects of the invention. Unless otherwise specified herein, scientific and technical terms used with this invention should have the meaning commonly understood by one of ordinary skill in the art. Furthermore, unless the context requires, singular terms should include the plural, and plural terms should include the singular. The article "a" is used herein for the grammatical purpose of referring to one or more (i.e., at least one) articles. For example, "an element" means one or more elements. Furthermore, claims using "comprising" allow other elements to be included within the scope of the claims; the invention also uses claims that use transitional phrases such as "consistently made of..." (i.e., other elements are allowed to be included within the scope of the claims if they do not significantly alter the operation of the invention) or "consisting of..." (i.e., only elements listed in the claims are allowed, excluding auxiliary elements or irrelevant activities generally related to the invention). Any of these three variations can be used to claim protection for this invention.

[0202] This document describes an apparatus for assays using a multi-well plate format having one or more of the following desired properties: (i) high sensitivity, (ii) large dynamic range, (iii) small size and weight, (iv) array-based multiplication capability, (v) automated operation, and (vi) the ability to process multiple plates. We also describe components and subsystems used in this apparatus, as well as methods of using the apparatus and subsystems. The apparatus and methods can be used with a variety of assay detection techniques, including but not limited to techniques for measuring one or more detectable signals. Some of these are suitable for electrochemiluminescence measurements, and in particular, embodiments suitable for use with multi-well plates (and assay methods using these plates) with integrated electrodes, such as those disclosed in U.S. Publication 2004 / 0022677 and U.S. Patent No. 7,842,246 to Wohlstadter et al., and U.S. Application 11 / 642,970 to Glezer et al., respectively.

[0203] In a preferred embodiment, an apparatus for performing luminescence analysis in a multi-well plate is provided. One embodiment includes a light detection subsystem and a plate handling subsystem, wherein the plate handling subsystem includes a light-shielding housing providing a light-free environment in which luminescence measurements can be performed. The housing includes a casing and a movable drawer housed within the casing. The casing also includes a casing top having one or more plate inlet holes through which plates can be lowered onto or removed from a plate transfer stage within the drawer (manually or mechanically). Before luminescence measurements are performed, a sliding light-shielding door in the casing seals the plate inlet holes, isolating ambient light. The casing also includes detection holes coupled to a light detector mounted on the casing top and one or more plate stackers mounted on the casing top above the plate inlet holes, wherein the plate stackers are configured to receive plates or to transport plates to a plate lifter within the movable drawer. The movable drawer includes a plate transfer stage for horizontally translating plates within the drawer to an area within the apparatus for performing specific analytical procedures and / or testing steps. The movable drawer also includes one or more board lifts with board lifting platforms that can be raised and lowered within the drawer, wherein the board lifts are located below one or more board inlet holes. The board translation stage is configured to position the board below the detection hole and above the board lift on the board lifting platform.

[0204] The device also includes a photodetector (e.g., via a light-shielding connector or baffle) mounted on a detection port on the top of the housing. In some embodiments, the photodetector is an imaging photodetector, such as a CCD camera, and may also include a lens. The photodetector can be a conventional photodetector, such as a photodiode, avalanche photodiode, photomultiplier tube, etc. Suitable photodetectors also include arrays of such photodetectors. Photodetectors that can be used also include imaging systems, such as CCD and CMOS cameras. The photodetector may also include lenses, light guiding devices, etc., for guiding light, focusing light, and / or imaging onto the detector. In some specific embodiments, the imaging system is used to image the emission from an array of combined regions in one or more wells of the assay plate, and the assay device reports the emission values ​​of the emission emitted by individual elements of the array. The photodetector is mounted on the top of the housing with a light-shielding seal. Other components of the device include plate contacts (e.g., for sensing ECL) for electrical contact with the plate and for supplying power to electrodes located below the wells of the photodetector.

[0205] Specific embodiments of the device of the present invention are shown in the figures. Figures 1(a)-(b) show front and rear views of the device 100 with a particular style of lid, respectively, and Figures 1(c)-(d) show corresponding front and rear views of the device without the lid, respectively. As shown, for example, in Figure 1(c), the device includes a light detection subsystem 110 and a board processing subsystem 120. More detailed figures are provided in Figures 2(a)-(b). The board processing subsystem 120 includes a light-shielding housing 130, which includes a housing 231 having a housing top 232, a bottom 233, a front portion 234, and a rear portion 235. The housing also includes a plurality of alignment features, and the housing is adapted to receive a movable drawer 240, which includes a movable drawer front portion and is composed of an integrally cast element. The walls of the movable drawer define a rigid xy subframe, 415 in Figure 4(d), including a plurality of mating alignment features. When the drawer is properly placed within the housing, the alignment features and mating alignment features engage and interlock, thereby aligning the drawer and its components with the components of the photodetector subsystem. When the alignment / mating alignment features are engaged, the weight of the movable drawer is supported by the top of the housing. The movable drawer 240 in the device 100 depicted in Figures 1(a)-(b) is... Figure 3 The interior is best shown in a partially open or closed position. The movable drawer 240, also shown in Figure 4(a), carries the various internal subsystems described in detail below, while in Figure 4(b), it is mounted within the housing 231, where the rear 235 and sides of the housing are omitted for clarity. Figure 4(c) shows the housing 231 with openings and alignment features 405, 406, and 407, the positions and dimensions of which are configured to receive the movable drawer 240.

[0206] In one embodiment, the board processing subsystem further includes a board sensor for detecting boards within the subsystem. Suitable board sensors include, but are not limited to, capacitive sensors, contact switches, ultrasonic sensors, weight sensors, or optical sensors, or combinations thereof.

[0207] Referring to Figure 2(a), the top of the housing 232 also includes one or more plate inlet (and outlet) holes, 236 and 237, through which plates are lowered onto or removed from the plate translation stage (manually or mechanically). A sliding light-shielding door (shown as 239 in Figure 2(c)) is used to seal the plate inlet holes 236 and 237 to isolate ambient light before luminescence measurements are performed. Additionally, the top of the housing includes an identifier controller for reading and processing data stored on the identifiers on the plates. In one embodiment, the identifier controller is a barcode reader (238) mounted above an opening in the top of the housing by means of a light-shielding seal, wherein the barcode reader is configured to read barcodes on plates placed on the plate translation stage within the housing. In a preferred embodiment, the barcodes on the plates are read as soon as the plates have been lowered into the drawer. In alternative or other embodiments, the plates include EEPROM or RFID, and the top of the housing and / or the drawer includes an identifier controller suitable for communicating with each of these identifiers. In other embodiments, the identifier controller may be provided independently of the device. In this embodiment, information stored in an identifier is transmitted to the device via a computer and / or a network attached to the device and / or manually entered via a user interface of a computer and / or a network, the identifier being attached to or associated with a board or a group of boards. For this information, see U.S. applications Serial Nos. 12 / 844,345 and 13 / 191,000, the disclosures of which are incorporated herein by reference.

[0208] The plate handling subsystem also includes a plate stacker mounted on the top 232 of the housing, above the plate inlet holes 236, 237, wherein the plate stacker is used to receive plates or transport plates to a plate lift. The plate handling subsystem may optionally include heating and / or cooling mechanisms (e.g., resistance heaters, fans, heat sinks, or thermoelectric heaters / coolers) to maintain the temperature of the subsystem below required conditions. It may also include humidity control mechanisms (e.g., humidifiers and / or dehumidifiers, or desiccant chambers) to maintain the humidity of the subsystem below required conditions.

[0209] A detailed view of the movable drawer of the board handling subsystem is shown in Figure 4. Referring to Figure 4(a), the drawer includes (i) a board lifting mechanism 400 with board lifting platforms 401 and 402 that can be raised and lowered; and (ii) a board translation platform 403 that translates the board in one or more horizontal directions, wherein the platform includes a board carriage 404 for supporting the board. The board carriage 404 preferably has an opening 420 to allow the board lift 400, positioned below the board carriage 404, to approach and lift the board, and the board translation platform 403 is configured to position the board below the detection aperture on the top 232 of the housing and below the photodetector within the photodetector system 110, and to position the board above the board lift 400. The board lifting platforms 401 and 402 of the board lift 400 preferably include anti-slip surfaces to prevent the board from moving on the board lifting platforms during movement within the device. The plate translation stage 403 has horizontal movement, for example, on a substantially horizontal plane or along the X and Y directions, to horizontally translate the plate within the drawer to one or more areas within the device for specific testing procedures and / or detection steps. In a non-limiting embodiment, as shown in FIG4(e), the plate translation stage 403 can move along a track 422 in a horizontal direction, and the plate carriage 404 can move in an orthogonal horizontal direction along a track 424 on the plate translation stage 403. In a preferred embodiment, the plate translation stage has two axes of motion, x and y, and motors coupled to the axes of motion allow the plate to move automatically on the stage.

[0210] The inclusions of the movable drawer 240 within the light-shielding housing 130 enhance the applicability and manufacturability of the device. To ensure proper alignment of the drawer 240 within the housing 231, and consequently, proper alignment of the subsystems within the drawer 240 with the light detection subsystem 110, the housing includes multiple alignment features, while the drawer's xy-mount includes multiple mating alignment features configured to engage and mesh with the alignment features of the housing. Figure 4(b) shows a cross-sectional view of the drawer 240, which is placed within the housing 231 having the rear portion 235 and the sides of the housing (omitted for clarity), and properly aligned with the light detection subsystem 110.

[0211] In a preferred embodiment, the alignment features of drawer 240 include a plurality of holes, while the corresponding alignment features on housing 231 include a plurality of pins sized to fit into the holes. As shown in FIG4(c), housing 231 preferably includes at least three alignment pins: pins 405 and 406 located on the front portion 234 of housing and pin 407 located on the opposite end of housing. Additional alignment features may be included in housing and drawer as needed. Preferably, the alignment features are positioned or aligned relative to the top of housing so that the weight of drawer 240 is supported by the top 232 of housing. The mating alignment features on drawer configured to engage and engage with alignment pins 405, 406, and 407 are shown in FIG4(d) as holes 408, 409, and 410 (in the embodiment shown in FIG4(d), alignment pin 405 engages and engages with hole 408, pin 406 engages and engages with hole 409, and pin 407 engages and engages with hole 410). Additionally, the drawer includes alignment latches 416 and 417 (shown in Figure 4(a)), which engage with mating alignment latches 418 and 419 (Figure 4(c)) to lock / unlock the drawer inside the housing.

[0212] Because alignment features 405-407 and 408-410 are positioned or aligned with the housing top 232, when the movable drawer 240 is inserted into the housing 231 guided by the XY frame 415, the weight of the drawer 240 and its components is supported by the housing top 232 after the drawer 240 is fully inserted into the housing 231. An advantage of this feature is that, since the photodetector system 110 is also mounted on the housing top 232, any alignment or alignment of the subsystems on the drawer 240 with the photodetector system 110 can be performed directly relative to the photodetector system 110, without considering any gaps or intervals between the drawer 240 and the housing top 232.

[0213] One or more additional engagement / locking features may be included in the housing and / or drawer, for example, as shown in FIG4(e), where a spring-loaded pin 411 is mounted to drawer 240 and configured to engage with a hole 412 positioned in the board carriage 403. In one embodiment, a solenoid is used to actuate the spring-loaded pin, such as pin 411. In the embodiment shown in FIG4(f), when the board carriage and board translation platform are aligned, as shown in FIG4(f), the alignment feature, pin 411, in the board translation platform engages with a corresponding locking feature, element 412, in the board carriage. These alignment and / or engagement features lock the board carriage in place to prevent damage to the components during, for example, shipping and / or installation.

[0214] In another preferred embodiment, as shown in Figures 4(c)-(d), the top of the housing includes an electrical connection contact mechanism 413, while the front of the drawer includes a mating electrical connection element 414, wherein the electrical connection and its mating are configured to engage and mesh with each other once the drawer is properly inserted into and aligned within the housing.

[0215] Referring to Figure 4(a), in a preferred embodiment, the plate carriage includes a carriage platform 404 and a plate locking mechanism for receiving and engaging an exemplary plate, hereinafter referred to as 426, placed on and in the carriage platform 404, as shown in Figures 5(a)-(b) (Figure 5(a) shows a diagram of a plate carriage with a locked plate 426, while Figure 5(b) shows a diagram of the same components with a visible plate locking mechanism engaging with the plate in the locked position). As shown in Figure 5(b), the outer edge of the plate follows the standard design conventions of multi-well plates and includes a skirt 522 that surrounds the plate wall and is lower than the plate wall (an enlarged view is shown in Figure 5(o)). In other words, the skirt 522 is located near the bottom of the plate 426. The plate locking mechanism is designed to push the outer edge of the skirt on two orthogonal sides of the plate against two corresponding physical restraints in the plate carriage, thereby clearly and reproducibly positioning the plate in the carriage. The plate locking mechanism is also designed to apply a downward physical force to a defined position on the top of the plate skirt to reproducibly and securely hold the plate in the vertical dimension.

[0216] A diagram of the plate carriage 404, the plate locking mechanism, and the plate 420 is shown in Figures 5(a)-(b). The sequence of operation of the plate locking mechanism is shown in Figures 5(c)-5(f) and is described below. In one particular embodiment, the plate carriage 404 supports a multiwell plate 426 (or a consumable having the same base surface and external physical geometry as a multiwell / microtiter plate configured for use in the apparatus described herein), the multiwell plate having at least first, second, third, and fourth sides, wherein the first and third sides are substantially parallel to each other, and the second and fourth sides are substantially parallel to each other. The plate carriage 404 defines a hole 420 that is substantially the same shape as the multiwell plate 426 but smaller in size, to support a skirt or flange 522 located around the multiwell plate 426. The plate carriage also includes first (501) and second (513) limiting surfaces that, when the plate 426 is fully locked, respectively define the horizontal position of the skirt 522 on the first and second sides of the multiwell plate. The plate locking mechanism can be moved from the open structure of the receiving plate 426 (best shown in Figures 5(i) and 5(j)) to the clamping structure, which locks the plate to the plate carriage (best shown in Figures 5(a) and 5(b)).

[0217] The plate locking mechanism includes (i) a first locking element (509) biased to a clamping position, the locking element being provided by a pedal 511, an actuating rod 510, and a spring 512, the spring preferably having a high elasticity. The pedal (511) is adapted to push a first side of the multi-well plate 426 toward a first stop 501 and a plate clamping arm (502) also biased to a clamping position by the spring 512, wherein the first locking mechanism (509) is connected to the plate clamping arm (502). The plate locking mechanism also includes (ii) a bracket (503) pivotally connected to the plate clamping arm (502) and adapted to push a second side of the plate 426 toward a second stop (513). The plate locking mechanism also includes (iii) at least one biasing clamp (515) positioned adjacent to the second stop (513) to clamp the skirt 522 of the multi-well plate 426 to the plate carriage 404, thereby preventing vertical movement. The clamp 515 engages with the skirt of the plate and applies a downward force to the skirt. The bracket (503) preferably includes at least two legs (504, 506), both of which contact the fourth side of the multiwell plate. At least one leg (504, 506) includes a ramp (507, 508) to apply a lateral force to the second stop and apply a downward force to the skirt of the multiwell plate (as shown in Figures 5(e)-(i)).

[0218] The first locking element 509 includes an actuating rod (510) biased by a spring (512) to a clamped position, extending beyond one edge of the plate carriage (as shown in Figure 5(c)). During plate loading and unloading, as the plate carriage 404 moves to align with the plate lift, the extension 510a of the actuating rod (510) is pushed against a physical stop within the housing, such as the rear wall of drawer 240 or the rear of housing 235, as clearly shown in Figure 5(d) where the rod 510 is not engaged and Figure 5(e) where the rod 510 is pushed. The physical stop pushes the extension 510a of the rod (510) into the carriage. It should be noted that when the plate carriage 404 moves to abut against the physical stop, both the rod 510 and the two bias clamps 515 are pushed. For clarity, Figures 5(d) and 5(i) only show the retraction of the rod 510. The movement of the lever (510) pushes the pedal 511 back toward the lever 510 to make room for the plate 426. As shown in FIG5(c), the pedal 511 is a cantilever arm attached to the lever 510 and has the ability to bend like a spring. When the lever 510 is pushed inward, the fulcrum 524, which is fixedly attached to the plate carriage 404, pushes the pedal 511 back, or moves in the direction of the arrow shown in FIG5(d). As best shown in FIG5(a), the fulcrum 526 can also be placed on the sheath 526 covering the first locking element 509. One end 528 of the plate clamping arm 502 is preferably pivotally connected to the lever 510, while its opposite end 530 is preferably pivotally connected to the plate carriage 404. The bracket 503 is pivotally connected to the plate clamping arm 502 at pivot point 531. As best shown in Figure 5(d), when the lever 510 is pushed inward, the pedal 511, the plate clamp arm 502, and the bracket 503 retract or leave the opening 420.

[0219] The advantage of pivotally connecting the bracket 503 to the plate clamping arm 502 is that the bracket 503 can rotate, preferably slightly relative to the plate clamping arm 502, so that the two legs 504 and 506 of the bracket 503 can contact the plate 426 during locking.

[0220] As described above, when the tray carriage 404 moves to abut against the physical limiter, the rod 510 and both bias clamps 515 are pushed. When the extension 515a of the bias clamp 515 is pushed inward, this action overcomes the force of the spring 532 and lifts the bias end 515b upward. When the bias end 515b is lifted to the open position, its dimensions are configured to fit the skirt 522 of the receiving plate 426, and when the bias clamp 515 is released, the spring 532 causes the bias end 515b downward and clamps it onto the skirt 522 to prevent the tray 426 from moving upward.

[0221] The device also includes an ejector (516) that releases the plate 426 from the locking mechanism. The ejector 516 has an extended actuating element (521), and a similar actuating rod (510) is also pushed against a limiter within the instrument when the plate carriage is aligned with the plate lifter, so that the ejector moves the multi-well plate 426 away from the second limiter 513. The ejector 516 is preferably spring-loaded by a spring 514, and it optionally includes an overtravel protector 534. When the ejector 516 is activated, it pushes the tray 426 away from the limiter 513, and when the ejector 516 is activated, the rod 510 and the bias clamp 515 also move to the open position so that the tray 426 can be pushed away from the limiter 513 and the bias end 515b. The overtravel protector 534 can be elastically deformable to absorb some of the ejector's movement. The movement of the carriage plate 404 away from the plate loading / unloading position (i.e., aligned with the plate lift) causes the movement of the flip bar (510) and the discharger (516) to reset the locking mechanism to the locking configuration.

[0222] Figures 5(i)-(m) illustrate the engagement of the multi-well plate 426 with the plate locking mechanism to lock the plate 426 within the plate carriage 404. Figure 5(i) is similar to Figure 5(d), showing the first locking element 509 with the pedal 511 retracted and the arm 502 / bearing 503 in the open position. In Figure 5(j), the locking mechanism remains disengaged and in the open position, allowing the multi-well plate 426 to be placed above the opening 420 within the plate carriage 404. In the opening configuration depicted in Figure 5(j), the pedal 511, clamping arm 502, bracket 503, and bias clamp 515 are biased away from the opening 420 to allow the plate 426 to be loaded within the plate carriage 404. As shown in Figure 5(j), the extensions 510a and 515a are pushed inward by the movement of the plate carriage 404 against a reverse limiting element, such as the back of the drawer 240 or the rear of the housing 235.

[0223] When plate 426 is placed in plate carriage 404 as shown in FIG. 5(k) and plate carriage 404 moves away from the reverse limiter, pedal 511 moves away from fulcrum 524 and outwards to push tray 426 and bias it against the first limiter 501. Plate clamp arm 502 also moves with rod 510, allowing bracket 503 to push tray 426 against the second limiter 513. As shown in FIG. 5(k), only leg 504 contacts tray 426; however, due to the pivot connection at pivot point 531, the second leg 506 automatically and quickly contacts tray 426 as bracket 503 rotates about pivot 531. As shown in FIG. 5(1), bias clamp 515, preferably spring-pressed by spring 532, engages the skirt 522 of multi-well plate 426 on the second side of plate, and bracket 503 also engages with skirt 522 and presses down on skirt 522. As described above, the legs 504 and 506 of the bracket 503 have ramps 507 and 508 at the angle shown. When the legs 504 and 506 push the tray 426, the ramps 507 and 508 contact the skirt 522 and push the tray 426 in two directions: toward the second stop 513 and downward. As shown in Figure 5(m), the biasing clamp 515 engages with the skirt 522.

[0224] In a preferred embodiment, the slide carriage 404 further includes an optical focusing mechanism used by optical sensors in the device (e.g., photodetectors within the light detection system 110 as described above) to measure contrast and focus. The optical focusing mechanism includes at least two, or preferably at least three, patterned surfaces at different heights relative to the slide carriage and thus relative to the target surface for focusing (i.e., the bottom of the well held in the 96-well plate 426 within the slide carriage 404). The invention includes a method for imagerizing a plurality of surfaces and calculating the magnitude and direction of image conditioning required to bring the target surface to focus based on the images. In one embodiment, a contrast value is calculated for the image of each surface, and a focus height is determined as a height at which a change is minimal or reduced to below a predetermined threshold compared to a change in height.

[0225] In one embodiment, the slide carriage includes at least three patterned surfaces, each at a different height relative to the slide carriage. Two alternative embodiments of the optical focusing mechanism are shown in Figures 6(a)-(b). In some preferred embodiments, the surfaces have patterns with varying transparency (e.g., patterns etched or cut into an opaque substrate, or patterned opaque ink or film printed on a transparent surface) so that the patterns can be imaged using light transmitted through the substrate. In an alternative embodiment, the surface / pattern is opaque, and the pattern is imaged using a light source that reflects light away from the surface.

[0226] The focusing mechanism includes at least patterned upper, middle, and lower surfaces spaced apart from the optical sensor, wherein the patterned middle surface and the target surface are substantially aligned with the same horizontal plane, and a first distance between the patterned upper and middle surfaces is substantially equal to a second distance between the middle and lower surfaces. The optical sensor and the patterned surfaces move relative to each other until the difference between a first pair of contrast values ​​between the upper and middle patterns and a second pair of contrast values ​​between the middle and lower patterns is less than a predetermined value of approximately ±2.0 in dimensionless units, as explained below. This difference can be ±3.0 or ±4.0, or as low as ±1.0. Higher contrast differences allow for easier but less precise focusing, while lower contrast differences produce more difficult but more precise focusing.

[0227] As shown in Figures 6(a)-(b), the mechanism preferably includes a plurality of patterned surfaces, for example, at least two and optionally three patterned surfaces (601-603), and the patterned surfaces include substantially the same pattern, for example, a grid pattern. The patterned surfaces are preferably clustered close to each other. In the embodiment shown in Figure 6(a), the mechanism also includes an unpatterned surface 604. Preferably, each patterned surface lies on a parallel plane. In a preferred embodiment, the height of the patterned middle surface is substantially equivalent to the focal position of the wells filled with a predetermined amount of fluid on the multi-well tray 426. The height of the patterned lower surface is approximately 0.25 mm below the patterned middle surface, while the height of the patterned upper surface is approximately 0.25 mm above the patterned middle surface. In one embodiment, the height of the patterned lower surface is approximately 4-4.75 mm above the plate carriage (i.e., above the carriage platform on which the plate is placed). Preferably, the patterned lower surface is positioned at a height approximately 4.5-4.7 mm above the slide plate; most preferably, the patterned lower surface is positioned at a height approximately 4.6-4.7 mm above the slide plate. The patterned middle surface is positioned at a height approximately 4.5-5.0 mm above the slide plate; preferably, approximately 4.7-4.9 mm above the slide plate; most preferably, approximately 4.7-4.8 mm above the slide plate. Furthermore, the patterned upper surface is positioned at a height approximately 4.75-5.10 mm above the slide plate; preferably, approximately 4.8-5.0 mm above the slide plate platform; most preferably, approximately 4.85-4.95 mm above the slide plate. It should be noted that any of surfaces 601, 602, and 603 can be a middle patterned surface, an upper patterned surface, or a lower patterned surface. In a preferred embodiment, the optical focusing mechanism is adjacent to the slide plate.

[0228] Therefore, the present invention provides a method for focusing an optical sensor onto a target surface, comprising the following steps: (a) providing at least patterned upper, middle, and lower surfaces 601-603, wherein the middle patterned surface and the target surface are at the same focal height, and wherein a first distance between the patterned upper surface and the middle surface and a second distance between the middle surface and the patterned lower surface are substantially equal; (b) obtaining a first contrast value difference between the patterned upper surface and the middle surface using an optical sensor; (c) obtaining a second contrast value difference between the patterned middle surface and the lower surface using an optical sensor; and (d) comparing the first contrast value difference and the second contrast value difference, and determining whether the target surface is in focus and / or determining the magnitude and direction of focus adjustment required to place the target surface in focus.

[0229] During operation, the plate translation stage 403 translates the plate carriage 404 to position the optical focusing mechanism above the contact mechanism, as shown in Figures 7(a)-7(c). This mechanism includes a light source, such as light outlets 725-728 shown in Figure 7(c). Light outlets 725-728 may be connected to a single light-emitting diode (LED), or each light outlet may have its own LED or other light source. The light source illuminates, and a beam of light is displayed on the underside of the optical focusing mechanism, more specifically below surfaces 601-603. Preferably, light outlets 725-728 provide uniform illumination to surfaces 601-603. The optical sensor or camera in the light detection subsystem 110 thus images the optical focusing mechanism, calculates the difference in contrast values ​​as described above, and determines whether the target is in focus and / or determines the magnitude and direction of focus adjustment required to bring the target surface into focus. Based on the calculation, the focus of the optical sensor is adjusted accordingly, manually or automatically, for example, by using a mechanized focus adjustment mechanism. Preferably, the method further includes the step of adjusting the distance between the optical sensor and the target surface, and the step of repeatedly obtaining the first and second contrast values ​​and comparing these contrast values ​​until the difference between the first and second contrast values ​​is less than a predetermined value. The appropriate calculation for determining the contrast value uses a region of interest (ROI) of the image covered by the dot pattern of the focal target, for example, surfaces 601, 602, or 603, or a portion thereof. The average and standard deviation of all pixels within the ROI are measured and determined. The average (AVG) and standard deviation (StDEV) of the contrast value (%CV) used to calculate the ROI are measured or determined.

[0230] %CV = (StDEV / AVG) x 100 Then, subtract %CV (high and low) from each ROI to generate the difference, which is then reported to the operator. %CV, as shown above, is a unitless or dimensionless value.

[0231] By comparing the ECL value related to defocus with the nominal value, a preferred predetermined value for the difference in %CV contrast value was experimentally determined to be ±2.0. The magnitude of this difference can be varied depending on the contrast function. A certain amount of defocus is acceptable without altering the ECL. A preferred value of ±2 is within this range. Smaller values, such as ±1.5 or ±1.0, are more precise but also more difficult to achieve during focusing operations. Larger values, such as ±3.0 or ±4.0, are less precise but easier to achieve. Those skilled in the art can balance accuracy and operational difficulty according to the teachings of this invention. A difference in contrast value between ±1.0 and ±4.0 is within the scope of this invention.

[0232] Other methods for calculating or determining contrast values ​​may also be used, such as in Eli Peli's "Contrast in Complex Images," published in the Journal of the Optical Society of America, Vol. 10, October 1990, pp. 2032-2040. This reference is incorporated herein by reference in its entirety.

[0233] Additionally, the slide 404 includes multiple reference elements. One reference element includes a conductive bottom surface 536 located on the bottom surface of the slide 404, as shown in FIG. 5(n), which is used during the installation of the device to train the positioning of the contact mechanism for contacting the bottom of the plate 426 held in the slide 404. The contact mechanism, described in more detail below, includes a series of spring-loaded contact elements and can be raised to contact the bottom surface of the plate 426, for example, to initiate an ECL measurement. As shown in FIG. 5(n), the conductive bottom surface 536 is located on the underside of the slide 404 and is positioned at the same height as the bottom of the plate when the plate 426 is locked in the slide 404. During the installation or adjustment of the device, the contact mechanism is raised until it reaches the height of the contact element contact surface 536, which is detected by electrically measuring the resistance drop between the contact elements, providing a signal during the ECL measurement that the contact elements have properly contacted the conductive surface 536 and will properly contact the bottom of the plate. This measured height is used to set the height of the contact mechanism to contact the plate 426 held in the plate carriage 404.

[0234] Furthermore, the plate carriage 404 includes another reference element (depicted in FIG. 5(c) as a semi-circular hole cut into the plate carriage 404, i.e., element 517-520). A light source in the contact mechanism, such as a light outlet or LED 722, emits light through each hole 517-522. The plate translation stage 403, moving on the horizontal plane as described above, positions each hole 517-522 above the light outlet 722 as shown in FIG. 7(c). The light emitted through each hole is imaged by a photodetector in the photodetector system 110 as a reference for the position of the plate carriage 404 relative to other components of the device in xy space of the horizontal plane. In a preferred embodiment, the reference element includes one or more notches or cuts located at both ends of the reference surfaces / stoppers (501) and (503), for example on the edge of the plate platform, as shown in FIG. 5(c). Advantageously, these elements may also be imaged to determine whether the plate is in the correct orientation.

[0235] Light outlets 722 and 725-728 are preferably illuminated by a single LED. Suitable LEDs can be connected to a light tube or waveguide before reaching the light outlets. Depending on the applied voltage, suitable LEDs can have different output intensities. In one example, as shown in Figure 7(h), LED 739 is connected to a multiplexer 738. Microprocessor 729 can instruct multiplexer 738 to apply a first voltage to LED 739 to activate light outlet 722, and a second voltage to LED 739 to activate light outlets 725-728. Alternatively, multiple LEDs can be used for the light outlets.

[0236] The board handling subassembly also includes one or more shipping locks to lock the board carriage in place during transport, as described above and best illustrated in FIG. 4(e). In a preferred embodiment, the shipping lock includes a solenoid drive pin 411 on the movable drawer 240, received in a hole 412 on the board translation platform 403. The board carriage 404 travels on tracks 422, 424 and preferably includes a clamp to lock the carriage in place. Further, the board carriage 404 includes a board orientation sensor, such as an accelerometer or electronic level, to ensure that the multi-well board 426 placed on the board carriage 404 is in the correct orientation.

[0237] The plate processing subassembly 120 also includes a plate contact mechanism comprising electrical contact probes mounted on a plate contact lift, as described above, for raising the probes to electrical contacts on the bottom of the multi-well plate 426, which in turn connect to electrodes within the wells of the plate. The contact probes are used to apply a potential to the electrodes in one or more wells of the multi-well plate 426. The plate contact mechanism is aligned with the imaging device so that the electrical contacts are directly below the well or set of wells of the imaging device and within its image domain. The contact mechanism is shown in Figures 7(a)-(b) and includes a contact mechanism platform 701 comprising four interrogation regions 702-705, each region comprising a pair of electrical contact probes to conduct a potential to the interrogation region. Preferably, the interrogation regions 702-705 are arranged in multiple quadrants or a 2x2 matrix. However, the interrogation regions can be arranged linearly or in any PxQ matrix, where P and Q are integers and can be distinct from each other. As discussed in more detail below, the multi-well plate 426 that can be used in the instrument 100 of the present invention can be arranged in an MxN matrix, wherein the MxN matrix is ​​larger than the PxQ matrix. As mentioned above, the PxQ matrix can be 12x8, 24x16, 48x32 wells or any number of wells.

[0238] The device also includes a controller operatively connected to a voltage source, which can be connected to one or more pairs of electrical contact probes and a multiplexer connected to the controller and the voltage source for selectively connecting the voltage source to the pair of electrical contact probes in a single interrogation area, or connecting the voltage source to multiple pairs of electrical contact probes in more than one interrogation area. A block diagram of the components of the controller is shown in Figure 7(h), including a microprocessor 729 connected to a power supply 730 and a digital-to-analog converter 731 connected to low-pass filters 732 and 733, a current monitor 734, another optional power supply 737, an analog-to-digital converter 736, and a multiplexer 738. The controller is also operatively connected to an LED 739, which is a component of the aforementioned contact mechanism.

[0239] The multiplexer 737, controlled by processor 729, guides the application of the potential as indicated above, depending on the type of board used in the instrument. If the multi-well board 426 is configured to analyze one well at a time, referred to herein as a single-well addressable board, where the wells of the board correspond to areas of the contact mechanism platform, the multiplexer 737 guides the selective application of the potential by electrically insulating each area and selectively applying the potential only in the first area. On the other hand, if the multi-well board is configured to analyze two or more wells at a time, referred to herein as a multi-well addressable board, then the multiplexer 737 guides the selective application of the potential by electrically connecting two or more areas and selectively applying the potential in these two or more areas. In one embodiment, the board includes a barcode containing board structure information, and the device 100 includes a barcode reader 238 that reads the board structure information and identifies the type of board located in the stacker crane.

[0240] In a preferred embodiment, the device includes a plurality of interrogation regions 702-705 mounted in a PxQ matrix. Preferably, the PxQ matrix is ​​a 2x2 matrix. The plurality of pairs of electrical contact probes on the plate contact mechanism platform 701 preferably include upright pins, such as spring-loaded pins. Furthermore, the device preferably also includes an optical sensor, such as a photodetector in the photodetector system 110, positioned above the platform 701. The platform 701 includes a first alignment mechanism comprising a light source, such as a light outlet 722 emanating from the platform towards the optical sensor to align the platform 701 relative to the optical sensor. In one embodiment, the light source (e.g., an LED or other type of bulb) is positioned below an aperture in the contact mechanism and emits light through the aperture, for example, through an aperture (722) located at the center of the platform (701) as shown in FIG. 7(c). The device preferably further includes a second alignment mechanism comprising a plurality of holes on the plate carriage frame (e.g., elements 517-520 shown in FIG. 5(c)), through which a light source 722 can be illuminated from the platform 701 and detected by an optical sensor to further align the plate carriage frame with the platform 701. The plurality of holes may be provided on at least two sides of the plate carriage frame (as described above). Furthermore, the device preferably includes a third alignment mechanism comprising a conductive surface on the plate carriage frame (e.g., surface 536 in FIG. 5(n)) such that when an electrical contact on the platform contacts the conductive surface, current flows between the electrical contacts on the platform to indicate a predetermined distance between the electrical contacts and the plate carriage. The device preferably includes a fourth alignment / focusing mechanism comprising patterned focusing targets (e.g., surfaces 601-603 in FIG. 6(a) and 6(b)), and the contact mechanism platform includes one or more light sources to allow light to pass through the pattern, as described above, to image the pattern. As described above, the light source may be a light source below the hole (722). Alternatively, multiple light sources (e.g., LEDs or other types of bulbs) can be used to generate a wider and more uniform light field, such as the four LEDs (725-728) in the embedded plate contact mechanism platform shown in Figure 7(c).

[0241] In a preferred embodiment, the apparatus is suitable for interrogating samples contained in a multi-well plate, wherein the multi-well plate comprises a plurality of wells arranged in an MxN matrix, and the apparatus includes a carriage frame configured to support the multi-well plate, wherein the carriage frame is movable relative to a contact mechanism platform, the platform comprising a plurality of interrogation areas, wherein each interrogation area includes at least one pair of electrical contact probes that apply a potential to at least one well. The apparatus also includes a controller and a multiplexer, the controller being operatively connected to a motor to move the carriage frame relative to the platform and operatively connected to a voltage source, wherein the voltage source can be connected to one or more pairs of electrical contacts, and the multiplexer being connected to the controller and the voltage source for selectively connecting the voltage source to the pair of electrical contact probes of a single interrogation area, or connecting the voltage source to at least one pair of electrical contact probes of more than one interrogation area. Preferably, the interrogation areas are arranged in a PxQ matrix, and the MxN matrix is ​​larger than the PxQ matrix, which may be a 2x2 matrix. Preferably, the specifications and dimensions of each interrogation area are set to interrogate one well on the multi-well plate 426.

[0242] Preferably, the electrical contact probes on the contact mechanism platform include multiple working electrode contact probes, which are selectively connected by a controller to a voltage source to determine the number of wells to be queried. In one embodiment, a working electrode probe is connected to a working electrode in one well, or a working electrode probe is connected to working electrodes in multiple wells. Unconnected working electrode probes can be electrically insulated in a multiplexer when not in use, thereby allowing multiple working electrode probes (e.g., four probes) to be used to apply a potential to multiple working electrodes in multiple wells, one well at a time (e.g., applying a potential to a group of four wells, one well at a time). The electrical contacts on the platform may also include multiple counter electrode probes electrically connected to at least one point ground. In one embodiment, the bottom electrical contacts of a multi-well plate connected to the counter electrode probes on the platform for multiple wells are electrically connected. Additionally, the bottom electrical contacts of a multi-well plate connected to the counter electrode probes on the platform for all wells are electrically connected. Furthermore, the bottom electrical contacts of a multi-well plate connected to the counter electrode probes on the platform for at least one well can be electrically insulated. The controller can simultaneously query PxQ or fewer wells.

[0243] Referring to Figures 7(c')-(g), the contact mechanism platform 701 includes multiple working contact probes 706-713 and counter contact probes 714-721. As shown in Figure 7(c'), if the controller 709 is configured to electrically connect two or more interrogation regions, then the instrument 100 selectively applies a potential in two or more regions, for example, regions 703 and 704, thereby applying the potential to the ends of working electrode contact probes 706 and 710 and 709 and 713, respectively, and connecting counter electrode contact probes 714-717 and 718-721. The connection between the counter electrode and plate 426 at platform 701 is discussed below. And as described below, only one working contact electrode and one counter contact electrode are required. The pairwise connections provide redundancy for the system so that an ECL signal is generated even when one electrode fails.

[0244] Additionally, if the switching mechanism is used to electrically insulate each region, the instrument will selectively apply a potential to the first region, for example, as shown in Figure 7(d), where region 703 is insulated and the potential is applied across working electrode contact probes 706 and 710. In one embodiment, all grounded counter electrode contact probes 714-717 and 718-721 are electrically connected to platform 701. As described below in conjunction with Figure 7(k), the counter electrode contact probes for each well are insulated from the counter electrode on the bottom of plate 426. In the embodiment shown in Figure 7(d), the well directly above region 703 has a counter electrode connected to counter electrode contact probes 718 and 719, but insulated from another counter electrode contact probe on platform 701. Furthermore, the counter electrode for each interrogation region can be insulated at platform 701.

[0245] Similarly, Figures 7(e)-(g) illustrate how the contact mechanism is configured to apply a potential to the first regions 702 (Figure 7(e)), 705 (Figure 7(f)), and 704 (Figure 7(g)), and the potential is applied across the working contact probes 707 and 712 (in Figure 7(e)), 708 and 711 (in Figure 7(f)), or 709 and 713 (in Figure 7(g)), respectively. Although the contact probes 714-717 and 718-721 are electrically connected to the platform 701, the contact probes in each interrogation region are insulated by the counter electrode on the well on the plate 426 directly above each interrogation region. Preferably, each contact probe is an independent spring-loaded contact element, such as a contact pin.

[0246] In a preferred embodiment, the multi-well plate 426 includes bottom electrical contacts on the bottom surface of the plate for each well, wherein the bottom electrical contacts are configured to contact the pair(s) of electrical contact probes on the platform 701. The bottom electrical contacts include counter electrode contacts connected to counter electrodes in the wells of the plate and working electrode contacts connected to working electrodes in the wells of the plate. Each well includes at least one working electrode and one counter electrode, which, depending on the form of the plate, may be electrically connected (engaged) or electrically insulated from working electrodes and counter electrodes in other wells of the plate.

[0247] A typical, non-limiting arrangement of the bottom electrical contact pattern is shown in Figures 7(i)-(l), where Figure 7(i) shows a pin contact configuration of platform 701 that is substantially similar to that of Figure 7(c'). Figure 7(k) shows the overlap of bottom electrical contacts beneath four exemplary wells covering inquiry areas 702-705. Each well has a bottom counter electrode 740 in an exemplary “Z” shape and two working electrodes 742 and 744. The bottom counter electrodes 740 are not electrically connected to each other, so the counter electrodes for each well or each inquiry area are independent or isolated at plate 426.

[0248] For region 703, the Z-shaped bottom counter electrode 740 is connected to counter electrodes 718 and 719. The bottom working electrodes 742 and 744 are connected to working electrodes 710 and 706, respectively.

[0249] For region 705, the Z-shaped bottom counter electrode 740 is connected to counter electrodes 720 and 721. The bottom working electrodes 742 and 744 are connected to working electrodes 711 and 708, respectively. Regions 702 and 704 are connected similarly.

[0250] The next electrical connection is to the interior of the well itself. As shown in Figure 7(1), each well in this example has a well working electrode 750 and well counter electrodes 752 and 754. Here, the well working electrode 750 is Z-shaped and connected to two bottom working electrodes 742 and 744, and the well counter electrodes 752 and 754 are connected to the bottom counter electrode 740.

[0251] For region 705, working electrodes 711 and 708 on platform 701 are connected to bottom electrodes 742 and 744 and well working electrode 750 for each well. Counter electrodes 720 and 721 on platform 701 are connected to bottom counter electrodes 740 and well counter electrodes 752 and 754 for each well. The Z-shape of bottom electrode 740 and well electrode 750 is designed to withstand sufficient electrical contact. Any shape can be used, and the invention is not limited to any particular shape.

[0252] As described above, each well and each inquiry zone has two working electrodes, for example, 708 and 711 for zone 705, and two counter electrodes, for example, 720 and 721 for zone 705. The two working electrodes and two counter electrodes are electrically connected to the well as shown above. Only one pair of working electrodes and counter electrodes is necessary to conduct the ECL potential to the well. The other pair is a spare in case of one or more electrode failures.

[0253] It should also be noted that in the examples described above in conjunction with Figures 7(i), 7(k) and 7(1), where each well can be queried separately, the working electrodes for each queried region and well are insulated at platform 701 and multiplexer 738, while the counter electrodes for each queried region and well are insulated at plate 426 and its bottom electrode and well electrode.

[0254] Figure 7(j) illustrates an example where four wells covering interrogation areas 702-705 can be simultaneously interrogated using contact pins or electrodes from the same platform 701. As shown, the multi-well plate 426 has a bottom working electrode 760 covering working electrodes 707, 708, and 709. The tray 426 also has a bottom counter electrode 762, covering at least counter electrodes 719, 720, 715, and 716. The bottom working electrode 760 and the bottom counter electrode 762 are electrically connected upwards to all four wells. Activating one or more working electrodes 707, 708, and 709 and one or more counter electrodes 719, 720, 715, and 716 provides an ECL potential for all four wells. Redundancy is also provided by multiple available working electrodes and counter electrodes.

[0255] According to one embodiment of the invention, the bottom of the plate includes internal electrical contact wires connected to bottom electrical contacts to conduct electrical potential into the well. In one embodiment, the bottom electrical contacts of at least one well are electrically insulated from the bottom electrical contacts of adjacent wells, and alternatively, the internal electrical contact wires for at least one well may be electrically insulated from the bottom electrical contacts for adjacent wells. Reference is made to U.S. Patent No. 7,842,246 and U.S. Application No. 2,004,002,2677 (both filed June 28, 2002, entitled "Assay Plate, Reading System, and Method for Luminescence Assay," which are therefore incorporated herein by reference), which disclose other embodiments of the bottom of the plate that may be inquired about by the contact mechanism disclosed herein.

[0256] Therefore, the present invention provides a method for interrogating samples contained in a multi-well plate having a well matrix of MxN, comprising the steps of: (a) providing a plate contact mechanism platform having a plurality of interrogation regions; (b) providing at least one pair of electrical contact probes (e.g., a working electrode contact probe and a counter electrode contact probe) for each interrogation region, wherein each interrogation region is adapted to interrogate a single well; (c) selectively applying a potential to: (i) one interrogation region to interrogate one or more wells simultaneously, or (ii) multiple interrogation regions to interrogate multiple wells; and (d) moving the multi-well plate relative to the platform to interrogate other wells. A single well can be interrogated, or MxN wells can be interrogated (where MxN is greater than a PxQ matrix). The method may further include step (e) controlling the potential applied in step (c) by selecting at least one positive active contact probe (e.g., a working contact probe) from the plurality of pairs of electrical contact probes on the platform to connect to the potential. Step (e) may further include electrically insulating at least one positive active contact probe from connection to the potential. The method may also include step (f), providing a bottom electrical contact on the bottom surface of the multi-well plate, and optionally step (g), electrically insulating at least one grounding probe (e.g., a counter electrode probe) from the bottom electrical contact. Alternatively, all grounding probes may be insulated from each other from the bottom electrical contact.

[0257] As described above, the device can be used to measure multi-well plates of two alternative types: single-well addressable plates (i.e., plates where one well is interrogated by the device at a time), and / or multi-well addressable plates (i.e., plates where one portion of the plate is interrogated by the device at a time, where the portion is a group of adjacent wells). Various types of multi-well plates, including single-well and multi-well addressable plates, are described in U.S. Patent No. 7,842,246 and U.S. Application No. 2,004,002,2677 (both filed June 28, 2002, entitled "Assay Plate, Reading System, and Method for Luminescence Assay Determination," which are therefore incorporated herein by reference). The plate of the present invention comprises, but is not limited to, a plate top, a plate bottom, a plurality of wells, a working electrode, a counter electrode, a reference electrode, a dielectric material, electrical connections, conductive vias, and assay reagents. The wells of the plate are defined by holes / openings in the plate top, while the plate bottom may be attached directly to or together with other components to the plate top, and the plate bottom may serve as the bottom of the wells. One or more assay reagents may be contained in wells and / or assay areas of the plate. These reagents may be fixed or placed on one or more surfaces of the well, preferably on the surface of the electrode, and most preferably on the surface of the working electrode. The assay reagents may be contained or positioned by features within the well; for example, a patterned dielectric material may confine or concentrate the fluid. The top of the plate preferably comprises an integral cast structure made of a rigid thermoplastic material such as polystyrene, polyethylene, or polypropylene. The bottom of the plate preferably comprises electrodes (e.g., working and / or counter electrodes) comprising carbon, preferably a carbon layer, more preferably a screen-printed layer of carbon ink. In another preferred embodiment, the bottom of the plate comprises electrodes composed of screen-printed conductive ink deposited on a substrate.

[0258] A single-well addressable board includes a board top having a top hole and a board bottom cooperating with the board top to define wells of the single-well addressable board. The board bottom includes a substrate having a top surface and a bottom surface. Patterned electrodes are provided on the top surface, and patterned electrical contacts are provided on the bottom surface. The electrodes and contacts are patterned to define a plurality of well bottoms of the single-well addressable board. The patterns within the well bottoms include: (a) working electrodes on the top surface of the substrate, wherein the working electrodes are electrically connected to the electrical contacts; and (b) counter electrodes on the top surface of the substrate, wherein the counter electrodes are electrically connected to the electrical contacts, but not to other counter electrodes in other wells of the single-well addressable board. Preferably, the electrodes and contacts of the single-well addressable board are individually addressable.

[0259] A multi-well addressable board includes a board top having a top hole and a board bottom cooperating with the board top to define wells of the multi-well addressable board. The board bottom includes a substrate having a top surface and a bottom surface. The top surface has patterned electrodes, and the bottom surface has patterned electrical contacts. The electrodes and contacts are patterned to define two or more independently addressable portions of two or more co-addressable assay wells. Each portion includes two or more wells, and: (a) co-addressable working electrodes on the top surface of the substrate, wherein each working electrode is electrically connected to each other and electrically connected to at least a first electrical contact; and (b) co-addressable counter electrodes on the top surface of the substrate, wherein each counter electrode is electrically connected to each other, but not to the working electrodes, and connected to at least a second electrical contact. In one embodiment, the independently addressable portions include less than 50% of the wells of the multi-well addressable board, more preferably less than 20% of the wells of the multi-well addressable board. The independently addressable portions may include a 4x4 well array or a 2x3 array of independently addressable portions. In addition, independently addressable portions may include one or more rows or columns of wells.

[0260] The single-well or multi-well addressable plate can be a 4-well plate, 6-well plate, 24-well plate, 96-well plate, 384-well plate, 1536-well plate, 6144-well plate, or 9600-well plate. Electrodes of any plate shape include carbon particles, and may also include printed conductive material. One or more electrodes include a plurality of assay zones formed thereon. The plurality of assay zones may include at least four assay zones, preferably seven zones, more preferably at least ten assay zones, and the plurality of assay zones may be defined by openings in one or more dielectric layers supported on the working electrode. The boards that can be used in this device are available from Meso Scale Discovery (Rockville, MD: www.mesoscale.com), including but not limited to the following multi-well addressable boards (Meso Scale Discovery catalog numbers): L15XA-3, L15XB-3, L15AA-1, L15AB-1, L15SA-1, L15SB-1, L15GB-1, L45XA-3, L45XB-3, N45153A-2, N45153B-2, N45154A-2, and N45154B-2; and the following single-well addressable boards (Meso Scale Discovery catalog numbers): L55AB-1, L55SA-1, L55XA-1, and L55XB-1.

[0261] Accordingly, the device measures the emission from a multi-well plate by the following steps: first, detecting the plate format in the device, for example, reading a barcode on the multi-well plate including structural information of the plate; aligning the contact mechanism and imaging device so that one or more interrogation areas are directly below or within the imaging area of ​​the imaging device; then guiding the selective potential application by (a) electrically isolating each interrogation area of ​​the contact mechanism and selectively applying a potential only to a first area (for a single-well addressable plate); or (b) electrically connecting two or more areas and selectively applying a potential to these two or more areas (for a multi-well addressable plate). If a multi-well addressable plate is used in the device, the imaging system and contact mechanism are aligned with interrogation areas corresponding to groups or portions of adjacent wells, for example, a group of four adjacent wells, and the device selectively applies voltage to all wells in the portion. The device then moves the plate using a plate translation stage to reset the contact mechanism and imaging system using other interrogation areas corresponding to other portions or groups of wells, and selectively applies voltage to the wells in those other portions. If a single-well addressable plate is used in the device, the imaging system and contact mechanism are aligned with the interrogation areas corresponding to groups or portions of adjacent wells, for example, a group of four adjacent wells, and the device selectively applies voltage to each well of the portion one at a time. Similarly, the plate is moved by means of a plate translation stage to reset the contact mechanism and imaging system using other interrogation areas corresponding to wells in other portions, to interrogate each well of the other portions one at a time.

[0262] In a particular embodiment, the device can measure light emission from a single-well addressable plate or a multi-well addressable plate, wherein the device includes: (i) Board format recognition interface for identifying board formats; (ii) A plate translation platform for holding a multi-well plate and translating the multi-well plate in the XY plane; (iii) Includes multiple contact probe plate contact mechanisms, the plate contact mechanisms being positioned below the plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift that can raise and lower the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) A voltage source that applies a potential to the plate via a contact probe; and (v) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism, wherein (a) The imaging system is configured to perform PxQ matrix imaging of a well, the plate contact mechanism is configured to contact the bottom contact surface associated with the matrix, and the plate translation stage is configured to translate the plate to position the matrix aligned with the imaging system and the plate contact mechanism. (b) The device is configured to sequentially apply voltage to each well in a matrix of single-well addressable plates and to perform matrix imaging; and (c) The device is configured to simultaneously apply voltage to each well in a matrix of multi-well addressable plates and to perform matrix imaging.

[0263] Preferably, the PxQ matrix is ​​a 2x2 well array. The imaging system can collect separate images for each consecutively applied voltage in each well of the matrix for a single-well addressable board. The board type identification interface may include a barcode reader, EPROM reader, EEPROM reader, or RFID reader; alternatively, the board type identification interface may include a graphical user interface configured to allow a user to input board type identification information.

[0264] Therefore, methods for measuring the emission from a single-well addressable plate or a multi-well addressable plate using such a device include: (a) Load the plate onto the plate translation platform; (b) Identify whether the board is a single-well or multi-well addressable board; (c) Move the plate translation stage to align the well matrix of the first PxQ with the plate contact mechanism and the imaging system; (d) A lifting plate contact mechanism so that the contact probe on the contact mechanism contacts the bottom contact surface associated with the well matrix of PxQ; (e) If the plate is a single-well addressable plate, then while forming an image, emission is generated in the PxQ matrix and imaged by sequentially applying voltage to each well in the group; (f) If the board is a multi-well addressable board, then while imaging the matrix, emission is generated and imaged in the PxQ matrix by simultaneously applying voltage to each well in the matrix; and (g) For the other PxQ matrices in the board, repeat steps (c) through (f).

[0265] The movable drawer may include a light source (e.g., an LED) located below the detection aperture and at an altitude below the plate translation stage. In one embodiment, the light source or multiple light sources are components of the plate contact mechanism. Regarding the optical focusing mechanism, as described above, one (or more) light sources in the contact mechanism are used in conjunction with the optical focusing mechanism to adjust the contrast and focus of the photodetector relative to the plate.

[0266] In other embodiments, one or more light sources may also be used in conjunction with a reference hole or window to correct for errors in plate alignment. Light from the light source passes through the reference and is imaged onto an imaging device to determine the correctness of the plate alignment. Advantageously, a plate formed by a plate bottom that mates with the plate top (e.g., a plate with a screen-printed plate bottom that mates with an injection-molded plate top, as described in co-pending U.S. applications 2004 / 0022677 and 2005 / 0052646) includes a reference patterned (e.g., screen-printed) or cut out on the plate bottom to correct for misalignment of the plate bottom relative to the plate top. In one particular embodiment, the plate top on such a plate includes a hole (e.g., in the outer frame of the plate top) aligned with a reference on the plate bottom to allow for reference imaging. Accordingly, imaging of light generated below the plate can be used to transfer the precise position of the plate to image processing software and also to provide camera focus verification. The plate can then be realigned using a two-axis positioning device. Therefore, the device can process the plate using a plate positioning method, which includes: (1) providing a plate with an optical path opening; (2) illuminating the plate from the bottom; (3) detecting the light coming through the optical path opening; and (4) optionally, realigning the plate.

[0267] In a preferred embodiment, the contact mechanism platform includes a first alignment feature 722, and the photodetector subsystem includes a camera positioned above the platform, which is adjustable relative to the first alignment feature. Preferably, the first alignment feature is a light source, such as an LED. The camera in the photodetector subsystem is adjustable relative to the alignment feature in the XY plane. The platform may also include multiple other alignment features, such as at least one other alignment feature in each quadrant, and the camera position is adjustable relative to each other alignment feature. Other alignment features may include light sources, such as LEDs. Thus, as described above, the device can use an optical focusing mechanism to determine proper alignment of the contact mechanism and the detection aperture by: (1) illuminating the contact mechanism alignment feature; (2) detecting light from the alignment feature; and (4) optionally, realigning the plate translation stage, the photodetector, and / or the contact mechanism. In a preferred embodiment, the device determines that the contact mechanism is properly aligned before contacting the plate, then confirms the position of the plate by detecting light from the optical path opening in the plate, and realigns the plate if necessary.

[0268] As shown in Figures 7(a)-(b), the height of the contact mechanism platform is adjustable because the platform also includes a shaft 723 driven by a gear mechanism 724. In one embodiment, the gear mechanism includes a worm gear. In a preferred embodiment, the platform includes a plate surface area sized to accommodate a micro-titer plate (e.g., a multi-well plate), and the platform also includes an overflow collection area surrounding the plate surface area to prevent the components of the drawer from being accidentally splashed by fluid that may be contained within the multi-well plate.

[0269] The light detection subsystem of the device includes a light detector that can be mounted to a detection hole on the top of the housing via a light-shielding connector or baffle. In some embodiments, the light detector is an imaging light detector, such as a CCD camera, and it also includes a lens. The light detection subsystem is shown in Figure 8(a). The subsystem includes a light detector housing 801 surrounding the light detector (not shown) and attached to the top of the housing via a cast member 802, which is bolted to the top of the housing above the detection hole. A clip or clamp 803 is provided above the cast member, which includes an θ adjustment mechanism as shown in Figure 8(b), comprising a screw 804 and a gear 805. The camera focusing mechanism is also configured to focus the camera manually or by means of mechanical elements or both along the x, y, and z directions when needed. The light detection subsystem also includes one or more light-shielding elements to prevent light leakage within the light detection subsystem or at the joint between the light detection subsystem and the top of the housing. For example, cast rubber or other compressible materials can be clamped between the connecting parts to prevent light leakage. Additionally, the photodetector housing includes one or more ventilation and / or cooling elements to cool the photodetector within the housing. In one embodiment, the housing includes an air inlet and an exhaust outlet, each located at opposite ends of the housing. Other vents may also be located within the housing. In a preferred embodiment, the air inlet is sized to match a cooling fan located within the housing.

[0270] A lens integrated into the camera provides a focused image of the light emitted by the plate within a light-shielding housing. A septum is sealed to the detection aperture in the top of the lens and housing, allowing the imaging system to image the light from the housing while keeping the housing in the light-shielding environment unaffected by ambient light. Suitable cameras for use in the imaging system include, but are not limited to, conventional cameras such as cinema cameras, CCD cameras, CMOS cameras, etc. The CCD camera can be cooled to low electronic noise. Preferably, the lens is a high numerical aperture lens, which can be made of glass or injection-molded plastic. The imaging system can be used to image one or more wells of the plate at a time. Because the size of the CCD chip and the imaging area are very well matched, the light collection efficiency for imaging light from a single well is higher than that for imaging a group of wells. The reduced size of the imaging area and the increased collection efficiency allow for the use of small and inexpensive CCD cameras and lenses while maintaining high detection sensitivity.

[0271] If high resolution is not required, measurement sensitivity can be improved by hardware regrading on the CCD during image acquisition, which effectively reduces electronic read noise per unit area. Preferred regrading depends on the field of view, magnification, and CCD pixel size. In a preferred embodiment, the photodetector comprises a camera with a CCD having 512x512 pixels, each pixel being 24x24 micrometers in size, for a total area of ​​12.3x12.3 mm, and the lens having an image magnification factor of 1.45x. For such a detector and lens combination, a 4x4 regrading is preferred, producing a superpixel size of approximately 100x100 micrometers, which translates to an object plane resolution of approximately 150 micrometers at the ECL electrodes. Regarding their low cost and size, it is particularly advantageous to use uncooled cameras or cameras with minimal cooling (preferably cooled to approximately -20°C, approximately -10°C, approximately 0°C, or higher). In a preferred embodiment, the optical detection subsystem includes a lens assembly consisting of a series of lens elements (904 and 905) designed to produce a focal length diagram of an imaging well, and a bandpass filter (903) located within the optical path of the lens assembly so that light passing through the filter is substantially perpendicular to the filter. Figure 9 In the illustrated embodiment, the camera provides a focal length map of the imaged well (901).

[0272] The top of the plate handling subsystem housing also includes a plate stacker mounted on the top of the housing above the plate inlet holes, wherein the plate stacker is configured to receive plates or transport plates to a plate lift. The plate stacker may include movable stacking nests for accommodating multiple plates and preventing plates from shifting on the instrument, thereby ensuring that each plate in the stacking nest is correctly introduced onto the plate lift. In one embodiment, the stacking nest can hold at least 5 plates, preferably at least 10 plates, and the stacking nest can accommodate a nest extension element for further increasing the nest's capacity. The plate lift includes plate detection sensors, such as capacitive sensors; the stacker may also include plate detection sensors, such as capacitance, weight, or optical sensors.

[0273] A method for performing measurements in a multi-well plate using this device is provided. The plate can be a conventional multi-well plate. Measurement techniques that can be used include, but are not limited to, techniques known in the art, such as cell culture-based assays, binding assays (including agglutination assays, immunoassays, nucleic acid hybridization assays, etc.), enzyme assays, colorometric assays, etc. Other suitable techniques will be apparent to those skilled in the art.

[0274] Methods for measuring the amount of an analyte also include techniques for measuring the analyte by detecting a marker that can be directly or indirectly attached to it (e.g., by using a marker-bound pair of the analyte). Suitable markers include those that can be directly observed (e.g., visually visible particles and markers that generate measurable signals, such as light scattering, optical absorption, luminescence, chemiluminescence, electrochemiluminescence, radioactivity, magnetic fields, etc.). Markers that can be used also include enzymes or other chemically reactive components that have chemical activity that generates measurable signals, such as light scattering, absorption, luminescence, etc. The formation of the product can be detectable, for example, due to differences in measurable properties such as absorption, luminescence, chemiluminescence, light scattering, etc., relative to the substrate. Some (but not all) measurement methods that can be used with the solid-bound method according to the invention may benefit from or require a washing step to remove unbound components (e.g., markers) from the solid phase.

[0275] In one embodiment, measurements performed using the apparatus of the present invention can utilize electrochemiluminescence-based assays, such as electrochemiluminescence-based immunoassays. High sensitivity, wide dynamic range, and selectivity of ECL are important factors in medical diagnostics. Commercially available ECL instruments have proven exceptional performance and are increasingly widely used due to factors including excellent sensitivity, dynamic range, accuracy, and tolerances for complex sample matrices. Categories that can be sensed to emit ECL (ECL-active classes) have been used as ECL markers, for example, (i) organometallic compounds where the metal is derived, for example, from Group VIII noble metals, including organometallic compounds containing ruthenium and those containing osmium, such as the tri-bipyridine-ruthenium (RuBpy) half-family, and (ii) luminol and related compounds. Categories involved in ECL markers during ECL processing are referred to herein as ECL co-reactants. Commonly used co-reactants include tertiary amines (e.g., see U.S. Patent No. 5,846,485), oxalates, and persulfates from RuBpy for ECL and hydrogen peroxide from luminol for ECL (e.g., see U.S. Patent No. 5,240,863). The light generated by ECL markers can be used as an indicator signal in diagnostic procedures (Bard et al., U.S. Patent No. 5,238,808, incorporated herein by reference). For example, ECL markers can be covalently bound to binders such as antibodies, nucleic acid probes, receptors, or ligands; the binding agents involved in the interaction can be monitored by measuring the ECL emitted by the ECL marker. Alternatively, the ECL signal from an active ECL compound can indicate a chemical environment (e.g., see U.S. Patent No. 5,641,623, which describes ECL assays and monitors the formation or destruction of ECL co-reactants). Further background information on ECL, ECL markers, ECL assays, and instruments for performing ECL assays can be found in U.S. Patent Nos. 5,093,268; 5,147,806; 5,324,457; 5,591,581; 5,597,910; 5,641,623; 5,643,713; 5,679,519; 5,705,402; 5,846,485; 5,866,434; 5,786,141; 5,731,147; and 6,066,448. ; 6,136,268; 5,776,672; 5,308,754; 5,240,863; 6,207,369; 6,214,552 and 5,589,136 and published PCT numbers WO99 / 63347; WO00 / 03233; WO99 / 58962; WO99 / 32662; WO99 / 14599; WO98 / 12539; WO97 / 36931 and WO98 / 57154, all of which are cited here.

[0276] In some embodiments, plates suitable for use in electrochemiluminescence (ECL) assays are disclosed in U.S. Patent No. 7,842,246. The apparatus of the present invention can use plates configured to detect ECL from one well or more wells at a time. As described above, ECL plates configured to detect one well or more wells at a time include electrodes and electrode connectors with a specially patterned design to allow electrical energy to be applied to the electrodes in only one well or more wells at a time. This apparatus may be particularly suitable for assays in plates containing desiccants and / or sealed wells, for example, as described in U.S. Application 11 / 642,970 by Glazer et al.

[0277] In one embodiment, the method includes: (a) guiding a plate to a plate stacker, (b) opening a light-shielding door, (c) lowering the plate from the plate stacker to a lifting platform on a plate translation trolley, (d) sealing the light-shielding door, (e) translating the plate so that one or more wells are below a photodetector, (f) detecting luminescence from one or more wells, (g) opening the light-shielding door, (h) translating the plate to a position below the plate stacker, and (i) lifting the plate onto the plate stacker. In a preferred embodiment, the method further includes reading a plate identifier on the plate and identifying the plate structure, translating the plate to a position where one or more wells are below a photodetector, optionally imaging one or more alignment features on a contact mechanism, then adjusting the position of the photodetector relative to the contact mechanism, and then selectively applying a potential to one or more interrogation regions according to the plate structure. The method may also include translating a plate carriage to a position where one or more other wells are below a photodetector and detecting luminescence from one or more other wells. The method may also optionally include applying electrical energy to electrodes within one or more wells (e.g., to induce electrochemiluminescence).

[0278] Experiments based on ECL multiplexing are described in U.S. patent applications 10 / 185,274 and 10 / 185,363, respectively, U.S. Publications 2004 / 0022677 and 2004 / 0052646; U.S. patent application 10 / 238,960, U.S. Publication 2003 / 0207290; U.S. patent application 10 / 238,391, U.S. Publication 2003 / 0113713; U.S. patent application 10 / 744,726, U.S. Publication 2004 / 0189311; and U.S. patent application 10 / 980,198, U.S. Publication 2005 / 0142033.

[0279] A method for testing biological agents using the apparatus described herein is also provided. In one embodiment, the method is a binding assay. In another embodiment, the method is a solid-phase binding assay (in one embodiment, a solid-phase immunoassay) and includes contacting the test component with one or more binding surfaces that bind an analyte of interest (or a binding competitor thereof) present in the test component. The method may also include contacting the test component with one or more detection reagents capable of specifically binding to an analyte of interest. Multiplexing binding assay methods according to preferred embodiments may involve several forms that are available in the prior art. Suitable assay methods include sandwich or competitive binding assay forms. Examples of sandwich immunoassays are described in U.S. Patents 4,168,146 and 4,366,241. Examples of competitive immunoassays include methods disclosed in U.S. Patents 4,235,601; 4,442,204; and 5,208,535 to Buechler et al. In one instance, small molecule toxins, such as marine toxins and fungal toxins, can be advantageously measured using competitive immunoassays.

[0280] Binding reagents, binding components of the binding surface, and / or bridging agents that can be used as probes include, but are not limited to, antibodies, receptors, ligands, haptens, antigens, epitopes, mimitopes, aptamers, hybridization partners, and intercalators. Suitable binding reagent components include, but are not limited to, proteins, nucleic acids, drugs, steroids, hormones, lipids, polysaccharides, and combinations thereof. The term "antibody" includes complete antibody molecules (including mixed antibodies assembled in vitro from antibody subunits), antibody fragments, and recombinant protein structures, including antigen-binding domains of antibodies (e.g., as described in Porter and Weir, J. Cell Physiol. 67 (Appendix I): 51-64, 1966; Hochman et al., Biochemistry 12: 1130-1135, 1973; incorporated herein by reference). The term also includes complete antibody molecules, antibody fragments, and antibody structures that have been chemically modified, for example, by introducing a marker.

[0281] The measurements used herein should be understood to include both quantitative and qualitative measurements, and include measurements performed for a variety of purposes, including but not limited to detecting the presence of an analyte, determining the amount of an analyte, identifying known analytes, and / or determining the species of unknown analytes in a sample. According to one embodiment, the amount of a first binding agent and a second binding agent bound to one or more binding surfaces can be expressed as a concentration value of the analyte in the sample, i.e., the amount of each analyte per volume of sample.

[0282] The analyte can be detected using an electrochemiluminescence-based assay. Electrochemiluminescence measurements are preferably performed using binding reagents immobilized or collected on the electrode surface. Particularly preferred electrodes include screen-printed carbon electrodes, which can be patterned on the bottom of specially designed cartridges and / or multi-well plates (e.g., 24, 96, 384-well plates, etc.). Electrochemiluminescence from the ECL markers on the carbon electrode surface is induced and measured using an imaging plate reader described in co-pending U.S. applications 10 / 185,274 and 10 / 185,363 (both filed June 28, 2002, entitled "Assay Plates, Reading Systems, and Methods for Luminescence Assays," which are incorporated herein by reference). Similar plates and plate readers are commercially available (MULTI-SPOT® and MULTI-ARRAY® plates and SECTOR® instruments, Meso Scale Discovery, Meso Scale Diagnostics Division, LLC, Rockville, MD).

[0283] In one embodiment, antibodies immobilized on electrodes within the plate can be used to detect selected biologics in a sandwich immunoassay format. In another embodiment, a microarray of antibodies patterned on an integrated electrode within the plate can be used to detect multiple selected biologics in a sandwich immunoassay format. Accordingly, each well contains one or more capture antibodies immobilized on the plate's working electrode and selectively in a dried form or as a separate component, e.g., within the kit, labeled detection antibodies, and all other reagents necessary for sample analysis, for positive and negative control.

[0284] The patents, patent applications, publications, and test methods referenced in this specification are incorporated herein by reference in their entirety. This invention is not intended to be limited to the specific embodiments described herein. In fact, various modifications of the invention, other than those described herein, will be apparent to those skilled in the art from the foregoing description and drawings. Such modifications are intended to fall within the scope of the claims.

[0285] Parts list Reference number Part name 100 devices 110 Photodetector system with photodetector 120-board processing system 130 light-shielding housing 231 casing 232 Top of Casing 233 bottom of the casing 234 Front of the housing 235 housing rear 236, 237 plates for introduction / extraction 238 barcode reader 240 movable drawers 400 plate lifting mechanism 401, 402 plate lifting platforms 403 Plate Translation Platform 404 Stainless Steel Plate Carriage with Opening 405, 406, 407 are aligned with the pins. 408, 409, 410 Alignment Holes 411 Spring-loaded pin Holes in 412 plate carriage 404 413 Electrical contact mechanism on housing tip 232 Matching electrical contact mechanism on drawer 414 415X-Y rack 416, 417 Alignment Locks 418, 419 Alignment Locks Opening in 420 carriage Orbits 422 and 424 426 multi-well plate 501 First Limiting Component 502 plate clamp arm 503 bracket 504 legs 506 legs 507 bevel 508 bevel 509 First Locking Element 510 Actuator Extension of the 510a actuator rod 511 pedal 512 spring 513 Second Limiting Component 514 spring 515 bias clip Extension of 515a bias clip The bias terminal of the 515a bias clip 516 Discharge Unit 522 skirt 524 fulcrums 526 sheath The ends of arms 528 and 530, and arm 502. 531 Pivot Point 532 Spring for bias clip 515 534 Over-migration Protector 536 conductive bottom surface 701 Platform Inquiry areas on platforms 702, 703, 704, and 705, specifically on platform 701. Working electrodes on platform 701 of 706, 707, 708, and 709 710, 711, 712, 713 Counter electrode on platform 701 (714, 715, 716, 717) 718, 719, 720, 721, 722 Alignment light output 723 axis 724 Gear Mechanism 725, 726, 727, 728 optical outputs 729 microprocessor 730 power supply 731DAC 732 low-pass filter 733 low-pass filter 734 Current Monitor 736ADC 737 power supply 738 Multiplexer 739LED 740 bottom electrode 742, 744 bottom working electrode 750 well working electrode Electrode pairs of wells 752 and 754 760 bottom working electrode 762 bottom electrode 801 Photodetector Housing 802 Cast Components 803 clips or fixtures 804 screws 805 Gear 901 imaging well 902 camera 903 bandpass filter 904 lens 905 lens

Claims

1. A method for focusing an optical sensor onto spaced-out platforms, comprising the following steps: a. Provide at least one patterned upper, middle and lower surface, wherein the patterned middle surface and the platform are aligned with each other, and a first distance between the patterned upper and middle surfaces is substantially equal to a second distance between the middle surface and the patterned lower surface; b. Obtain the first contrast difference between the patterned upper and middle surfaces using an optical sensor; c. Obtain the second contrast difference between the patterned middle and lower surfaces using an optical sensor; d. Compare the difference between the first and second contrast values.

2. The method of claim 1 further includes step (e) adjusting the distance between the optical sensor and the platform, and repeating steps (b) to (d) until the difference between the first and second contrast values ​​is less than a predetermined value.

3. The method of claim 1, wherein the patterned middle surface is substantially aligned with a plane at the same level as the bottom surface of the tray, the tray carrying at least one sample to be interrogated by an optical sensor.

4. The method of claim 3, wherein the platform comprises a plurality of electrodes that contact the bottom surface of the tray to conduct current to the at least one sample.

5. The method of claim 1, wherein the patterned upper surface, middle surface and lower surface are located on parallel planes.

6. The method of claim 1, wherein the patterned upper, middle, and lower surfaces comprise substantially the same pattern.

7. The method of claim 6, wherein the same pattern includes a grid.

8. The method of claim 2, wherein the predetermined value is less than about ±4.

0.

9. The method of claim 8, wherein the predetermined value is less than about ±3.

0.

10. The method of claim 9, wherein the predetermined value is less than about ±2.

0.

11. The method of claim 1, wherein the distance between the patterned intermediate surface and the platform is between about 4 mm and about 4.75 mm.

12. The method of claim 11, wherein the distance is between about 4.5 mm and about 4.7 mm.

13. The method of claim 12, wherein the distance is between about 4.6 mm and about 4.7 mm.

14. The method of claim 1, wherein the patterned upper surface, middle surface and lower surface are located adjacent to each other.

15. The method of claim 1, wherein the patterned upper, middle and lower surfaces are located in one quadrant.

16. The method of claim 1, wherein the patterned upper, middle, and lower surfaces are illuminated by a light source positioned opposite the optical sensor.

17. The method of claim 1, wherein the optical sensor comprises a camera, a CCD sensor, or a CMOS sensor.

18. A focusing mechanism for an optical sensor, comprising at least a patterned upper surface, a middle surface, and a lower surface spaced apart from the optical sensor; The patterned mid-surface is aligned with the target surface that needs to be focused by the optical sensor and the patterned mid-surface. The first distance between the patterned upper and middle surfaces is substantially equal to the second distance between the middle surface and the patterned lower surface. The optical sensor and the patterned surface move relative to each other until the difference between the first and second contrast values ​​is less than a predetermined value; and The light source is positioned such that light passes through patterned upper, middle, and lower surfaces and is directed toward the optical sensor.

19. The focusing mechanism of claim 18, wherein the target surface includes a reference plane of the platform that selectively conducts current to the sample being interrogated by the optical sensor.

20. The focusing mechanism of claim 19, wherein the patterned middle surface is aligned with the reference surface of the platform by a predetermined amount.

21. The focusing mechanism of claim 20, wherein the distance is between about 4 mm and about 4.75 mm.

22. The focusing mechanism of claim 21, wherein the distance is between about 4.5 mm and about 4.7 mm.

23. The focusing mechanism of claim 22, wherein the distance is between about 4.6 mm and about 4.7 mm.

24. The focusing mechanism of claim 18, wherein the target surface includes the bottom surface of a tray holding at least one sample to be interrogated by an optical sensor.

25. The focusing mechanism of claim 24, wherein the patterned middle surface and the bottom surface of the tray are substantially aligned with the same horizontal plane.

26. The focusing mechanism of claim 18, wherein the patterned upper surface, middle surface and lower surface are located adjacent to each other.

27. The focusing mechanism of claim 18, wherein the patterned upper, middle and lower surfaces are located in one quadrant.

28. The focusing mechanism of claim 19, wherein the optical sensor comprises a camera, a CCD sensor, or a CMOS sensor.

29. The focusing mechanism of claim 18, wherein the patterned upper, middle and lower surfaces are located on parallel planes.

30. The focusing mechanism of claim 18, wherein the patterned upper, middle, and lower surfaces comprise substantially the same pattern.

31. The focusing mechanism of claim 30, wherein the same pattern includes a grid.

32. An instrument for performing luminescent analysis in a multi-well plate, the instrument comprising a light detection subsystem and a plate processing subsystem, wherein the plate processing subsystem comprises: (a) A light-shielding enclosure, comprising a housing and a movable drawer, wherein (x) The housing includes a housing top, a housing front, one or more plate inlet holes, a detection hole, a sliding light-shielding door for sealing the plate inlet holes, and multiple alignment features, wherein the housing is adapted to receive a movable drawer, and (y) Movable drawers, including: (i) an xy subframe comprising a plurality of mating alignment features configured to engage and mesh with the plurality of alignment features to align a movable drawer within the housing relative to the photodetector subsystem, wherein the weight of the movable drawer is supported by the top of the housing; (ii) One or more plate lifts having a plate lifting platform that can be raised and lowered, wherein the position of the one or more plate lifts is below the plate inlet hole; (iii) A plate translation platform for translating a plate along one or more horizontal directions, wherein the platform includes a plate carriage for supporting the plate, the plate carriage having an opening allowing a plate lifter positioned below the plate carriage to approach and lift the plate, and the plate translation platform is configured to position the plate below a detection hole and above the plate lifter; and (b) One or more plate stackers mounted on top of the housing, above the plate inlet holes, wherein the plate stackers are configured to receive plates or transport plates to a plate lifter; and The photodetector subsystem includes a photodetector mounted on the top of the housing and integrated into a detection aperture with a light-shielding seal.

33. The instrument of claim 32, wherein the xy subframe is mounted on top of the housing.

34. The instrument of claim 32, wherein the plurality of alignment features includes a set of alignment pins always distributed within the housing, and the plurality of mating alignment features includes a set of holes configured to engage with the set of alignment pins.

35. The instrument of claim 32, wherein the housing includes a plurality of electrical connectors, and the drawer includes a plurality of mating electrical connectors configured to contact and engage with the plurality of electrical connectors.

36. The instrument of claim 32, wherein one or more plate stackers are movable.

37. The instrument of claim 32 further includes a plate stacker extension element configured to accommodate a plurality of plates, wherein the plate stacker extension element is placed on top of the plate stacker.

38. The instrument according to claim 37, wherein the height of the plate stacker extension element is adjustable.

39. The instrument of claim 38, wherein the plate stacker extension element is configured to accommodate up to 20 plates.

40. The instrument of claim 38, wherein the plate stacker extension element is configured to accommodate up to 10 plates.

41. The instrument of claim 38, wherein the plate stacker extension element is configured to accommodate up to 5 plates.

42. The instrument of claim 32, wherein the board processing subsystem further includes a board sensor configured to detect boards in the subsystem.

43. The instrument of claim 42, wherein the plate sensor comprises a capacitive sensor, a contact switch, an ultrasonic sensor, a weight sensor, or an optical sensor.

44. The instrument of claim 32, wherein the drawer further comprises a plurality of overflow collection mechanisms adjacent to one or more plate lifts and / or plate translation platforms.

45. The instrument of claim 44, wherein the overflow collection mechanism includes a drip protection device.

46. ​​The instrument according to claim 32, wherein the plate lifting platform includes an anti-slip surface.

47. The instrument of claim 32, wherein one or more plate lifts comprise two adjacent lifting platforms connected by a cross mechanism for raising and lowering the lifting platforms.

48. The instrument of claim 32, wherein the plate translation stage includes a locking mechanism configured to prevent movement of the stage.

49. The instrument of claim 32, wherein the plate handling subsystem further comprises one or more plate orientation sensors.

50. The instrument of claim 49, wherein one or more plate orientation sensors are placed on or connected to the plate carriage, the one or more plate stackers and combinations thereof.

51. The instrument of claim 32, wherein the photodetector subsystem includes a camera and a camera focusing mechanism for focusing the camera in the x, y, z, and θ directions.

52. The instrument of claim 51, wherein the camera focusing mechanism includes manually adjustable elements.

53. The instrument of claim 52, wherein the manually adjustable element comprises a knob.

54. The instrument of claim 51, wherein the photodetector subsystem further comprises a motor configured to drive the camera focusing mechanism.

55. The instrument of claim 51, wherein the photodetector subsystem comprises a photodetector subsystem housing mounted to the board processing subsystem.

56. The instrument of claim 55, wherein the light detection subsystem housing includes a clip secured to the top of the light-shielding outer housing.

57. The instrument of claim 56, wherein the clip further comprises one or more light-shielding materials configured to prevent light leakage from the light-shielding housing.

58. The instrument of claim 51, wherein the housing of the light detection subsystem further includes a light shield surrounding the camera.

59. The instrument of claim 51, wherein the camera includes a lens having an F-number in the range of 1.3 to 1.

8.

60. The instrument of claim 51, wherein the housing includes an internal cooling fan, an air inlet located on a first side of the housing, and an exhaust port located on a side of the housing opposite to the first side.

61. An instrument for performing luminescence analysis on a multi-well plate, the instrument comprising a plate processing subsystem, the plate processing subsystem comprising a plate carriage for supporting the multi-well plate, wherein the plate carriage comprises a frame and a plate locking mechanism, the plate locking mechanism comprising: (a) Plate carriage flange; (b) A plate clamping arm perpendicular to the flange, including a proximal end and a distal end relative to the flange, wherein the proximal end of the arm is attached to the frame and the distal end of the arm is rotatable on the xy plate, and the arm also includes an upper clamp including an inclined surface configured to engage with the plate. (c) A plate positioning element, comprising a rod, a pedal, and a spring, wherein the rod is perpendicular to the arm, parallel to the flange, and attached to the distal end of the arm by means of the spring, while the pedal is attached to the rod at an angle; and (d) A plate wall parallel to the arm, perpendicular to the positioning element and the flange, and disposed between the two, the plate wall including (i) a lower plate clamp for engaging with the skirt of the multi-well plate, and (ii) a lower plate clamp ramp configured to push the lower plate clamp toward the skirt.

62. The instrument of claim 61, wherein the plate wall further includes a plate discharge element configured to disengage the lower plate clamp from the skirt.

63. A method for embedding a multi-well plate in the instrument of claim 61, comprising: (a) Place the board on the frame; (b) The spring of the compression plate positioning element pushes the pedal against the plate toward the flange and causes the arm to rotate toward the plate in the xy plane; (c) Make the upper clamp contact the plate, thereby pushing the plate against the carriage wall; (d) Make the lower plate clamp contact with the skirt, thereby locking the plate in the carriage.

64. An instrument for performing luminescence analysis in a multi-well plate, the instrument comprising a plate processing subsystem and a plate locking mechanism, the plate processing subsystem comprising a plate carriage supporting the multi-well plate. The multi-well plate has at least a first, second, third, and fourth side surface, with the first and third side surfaces being substantially parallel to each other, and the second and fourth side surfaces being substantially parallel to each other. The plate carriage defines a hole whose shape is basically the same as that of the multi-well plate and whose size is smaller than that of the multi-well plate to support the flange located around the multi-well plate. The plate carriage also includes a first limiting member (501) and a second limiting member (513) corresponding to the first and second sides of the multi-well plate, respectively. The plate locking mechanism can be moved from an open configuration for receiving a multi-well plate to a clamping configuration for locking the multi-well plate to the plate carriage. The plate locking mechanism includes a first locking element (509) biased to a clamping position and a plate clamping arm (502) biased to a clamping position. The first locking element has a pedal (511) adapted to push a first side of the multi-well plate toward a first stop. The plate clamping arm has a bracket (503) pivotally connected to the plate clamping arm (502) and adapted to push a second side toward a second stop (513). The first locking mechanism (509) is connected to the plate clamping arm (502), and... The plate locking mechanism includes at least one bias clamp (515) positioned adjacent to the second limiter (513) to clamp the skirt of the multi-well tray to the plate carriage.

65. The instrument of claim 64, wherein the bracket (503) comprises at least two legs (504, 506), both of which contact the fourth side of the multi-well tray.

66. The instrument of claim 65, wherein at least one leg (504, 506) includes an inclined plane (507, 508) to apply force perpendicular to the plane of the multi-well tray.

67. The instrument of claim 64, wherein the first locking element comprises an actuating rod (510) biased to a clamping position by a spring (512).

68. The instrument of claim 67, wherein the pedal (511) is attached to the actuating rod (510), and the plate carriage has a third limiting member to push the pedal toward and away from the multi-well tray when the actuating rod is moved.

69. The instrument of claim 67, wherein the pedal and plate clamp arm are retracted into the opening configuration.

70. The instrument of claim 61 further includes a discharge device that moves the multi-well tray away from the first or second limiting member.

71. The instrument of claim 70, wherein the discharge device is spring-loaded.

72. The instrument of claim 70, wherein the discharge device includes an over-travel protector.

73. An apparatus for measuring light emitted from a multi-well plate, the plate type of which is selected from a group consisting of single-well addressable plates or multi-well addressable plates, the apparatus comprising: (i) Board type identification interface for identifying board type; (ii) A plate translation platform for holding and translating a multi-well plate in the xy plane; (iii) A plate contact mechanism including multiple contact probes, the plate contact mechanism being positioned below the plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift capable of raising and lowering the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) Applying a potential to the plate via a voltage source using contact probes; and (v) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism, wherein (a) The imaging system is configured to image the PxQ matrix of the well, the plate contact mechanism is configured to contact the bottom contact surface associated with the matrix, and the plate translation stage is configured to translate the plate to a position where the matrix is ​​aligned with the imaging system and the plate contact mechanism. (b) The device is configured to sequentially apply voltage to each well in a matrix of single-well addressable plates and to image the matrix; and (c) The device is configured to simultaneously apply voltage to each well in a matrix of multi-well addressable plates and to image the matrix.

74. The apparatus of claim 73, wherein the PxQ matrix is ​​a well of a 2x2 array.

75. The apparatus of claim 74, wherein the imaging system collects separate images of the voltage of each well in a matrix of continuously applied single-well addressable plates.

76. The apparatus of claim 73, wherein the board type identification interface comprises a device selected from the group consisting of a barcode reader, an EPROM reader, an EEPROM reader, or an RFID reader.

77. The apparatus of claim 73, wherein the board type identification interface includes a graphical user interface configured to allow a user to input board type identification information.

78. A method for measuring luminescence from a multi-well plate, the plate type of which is selected from a group consisting of single-well addressable plates or multi-well addressable plates, the apparatus comprising: (i) Board type identification interface for identifying board type; (ii) A plate translation platform for holding and translating a multi-well plate in the xy plane; (iii) A plate contact mechanism including multiple contact probes, the plate contact mechanism being positioned below the plate translation stage and within the movement range of the stage, wherein the mechanism is mounted on a contact mechanism lift capable of raising and lowering the mechanism so that when the plate is on the translation stage, the probes may or may not contact the bottom contact surface of the plate. (iv) Applying a potential to the plate via a voltage source using contact probes; and (iv) An imaging system positioned above the plate translation stage and perpendicularly aligned with the plate contact mechanism. The method includes, (a) Load the plate onto the plate translation platform; (b) Identify whether the board is a single-well or multi-well addressable board; (c) Move the plate translation stage to align the first PxQ matrix of the well with the plate contact mechanism and the imaging system; (d) A lifting plate contact mechanism so that the contact probe on the contact mechanism contacts the bottom contact surface associated with the PxQ matrix of the well; (e) If the plate is a single-well addressable plate, then while the group is being imaged, light emission is generated in the PxQ matrix and imaged by sequentially applying voltage to each well in the group; (f) If the board is a multi-well addressable board, then while the matrix is ​​being imaged, light emission is generated and imaged in the PxQ matrix by simultaneously applying voltage to each well in the matrix; and (g) For the other PxQ matrices in the board, repeat steps (c) through (f).

79. The method of claim 78, wherein the PxQ matrix is ​​a 2x2 array of wells.

80. The method of claim 78, wherein the imaging system collects separate images of the voltage of each well in a matrix of continuously applied single-well addressable plates.

Images