JP2016538573A - Multiwell plate and method of using the same - Google Patents

Abstract

A plate is provided with a mechanism that facilitates determination of the amount of fluid added to or removed from wells in the plate. Other embodiments are also described. [Selection] Figure 3C

Description

Related Applications This is a US provisional application filed 61/905865 filed on 19 November 2013, 61/929086 filed on 19 January 2014, and filed on 7 March 2014. Claims the priority of the 61/949272 Paris Convention and the benefit of Section 120 of the US Patent Act. The contents of the provisional application described above are cited in the present specification.

Background Plates with multiple wells holding samples for observing chemistry, cells or other biological material are known in the prior art. Typically, such plates have a 24: 4 (6x6), 96 (8x12), 384 (16x24) or 1536 (32x48) well because the aspect ratio is 3: 2, and a typical 96 well plate is 128 mm wide and 86 mm wide, respectively, the footprint and bottom outer flange of the 96 well plate are described in ANSI / SBS 1-2004 and ANSI / SBS 3-2004, respectively.

  Such multiwell plates (sometimes also referred to as microwell plates or microtiter plates, depending on the volume of the well) are typically composed of plastic (eg polystyrene, polypropylene or polycarbonate or a combination of such materials), and in some cases the plate's Glass is also built into the bottom. In many applications, the bottom of the well transmits the frequency of light used to observe the sample. The shape of the well and the shape of the bottom of the well and the size of the well with respect to depth, height and total volume will vary depending on the particular use of the plate.

  Examples of commercial suppliers of such plates include Perkin-Elmer (see http://www.perkinelmer.com/CMSResources/Images/44-73879SPC_MicroplateDimensionsSummaryChart.pdf), Sigma-Aldrich (http://www.sigmaaldrich.com /labware/labware-products.html?TablePage=9576216) and Thermo-Scientific (see http://www.thermoscientific.com/ecomm/servlet/productscatalog_11152__81996_-1_4).

  One area where such plates are widely used is high-throughput screening (binding assays for antigens etc.) to test compounds in drug development.

  In many cases, in high-throughput screening and other applications, an automated machine, for example, dispenses a portion of a well by distributing fluid simultaneously to the wells of an 8-well row of a 96-well plate or to all wells of a 384-well plate. Or all, used to dispense a volume of liquid simultaneously. However, if the amount of liquid added to a particular well is incorrect, this fact will not be apparent until the entire experiment is complete, and the particular well with the wrong amount of fluid added will not be identified In some cases, it may be necessary to discard the results from the entire plate.

BRIEF DESCRIPTION OF THE DRAWINGS In accordance with one embodiment of the present invention, a plate comprising a first substantially planar surface, a second substantially planar surface, at least one well and at least one signal provider, wherein the first substantially planar surface is: At least one first opening defined therein, wherein the second substantially planar surface is substantially parallel to the first substantially planar surface, and the second substantially planar surface is the first substantially planar surface. At least one well is defined in the plate, and the at least one well has a second opening corresponding to and aligned with one of the at least one first opening. The at least one well comprises a sidewall and a bottom, and the at least one well extends from the first substantially planar surface toward the second substantially planar surface; The well is displaceable away from the first substantially planar surface, and the at least one signal provider is functionally associated with the at least one well and the displacement of the at least one well away from the first surface A plate is provided that can provide a signal in response to.

  In certain embodiments, the first surface comprises a plurality of first openings defined therein, the plurality of wells are defined in the plate, and each well is a plurality of second defined in the first surface. A second opening corresponding to and aligned with the first one of the openings, each well including a sidewall and a bottom, each well from the first substantially planar surface to the second substantially planar surface; It extends towards the surface. In some embodiments, each of the plurality of wells is displaceable away from the first substantially planar surface.

  Also according to one embodiment of the present invention is a multi-well plate comprising a first substantially planar surface, a second substantially planar surface, a plurality of wells and at least one signal provider, wherein the first substantially planar surface is A plurality of first openings defined therein, wherein the second substantially planar surface is substantially parallel to the first substantially planar surface, wherein the second substantially planar surface is from the first substantially planar surface; A plurality of wells are defined in the plate, each well comprising a second opening corresponding to and aligned with one of the first openings defined in the first surface; Each of the wells includes a sidewall and a bottom, each well extending from the first substantially planar surface toward the second substantially planar surface such that each of the wells is spaced from the first substantially planar surface. Can be displaced to at least one One signal provider provides a multi-well plate that is functionally associated with a plurality of wells and can provide a signal in response to displacement of at least one well away from the first surface.

  In some embodiments, the first and second surfaces are spaced apart by a plurality of sidewalls extending between the first and second surfaces.

  In some embodiments, all well movements are connected so that the signal provider can provide a single signal in response to any one or more well displacements. In some embodiments, several well movements connect to two or more groups and at least one signal provider comprises a multi-signal provider, each responding to one displacement of the group. Can provide a signal. In some embodiments, several well movements connect to two or more groups and the signal provider can provide separate signals in response to one displacement of each group.

  In some embodiments, the at least one signal provider comprises a plurality of signal providers, each associated with one well of the plurality of wells, each of the plurality of wells being away from the first surface. Each of the plurality of signal providers can provide a signal in response to displacement of one of the plurality of wells associated therewith.

  In certain embodiments, the at least one signal provider includes 300 milligrams of material in a well, 250 milligrams of material, 200 milligrams of material, 150 milligrams of material, 100 milligrams of material, 75 milligrams of material, 50 milligrams of material, 45 Milligram substance, 40 milligram substance, 35 milligram substance, 30 milligram substance, 25 milligram substance, 20 milligram substance, 15 milligram substance, 10 milligram substance, 5 milligram substance, 4 milligram substance, 3 A signal can be provided in response to the placement of milligrams of material, 2 milligrams of material or 1 milligram of material, 500 micrograms (μg) of material, 300 μg of material, 200 μg of material or 100 μg of material.

  In certain embodiments, at least one signal provider is 300 microliters (μl), 250 μl fluid, 200 μl fluid, 150 μl fluid, 100 μl fluid, 75 μl fluid, 50 μl fluid, 45 μl fluid, 40 μl in a well. Fluid, 35 μl fluid, 30 μl fluid, 25 μl fluid, 20 μl fluid, 15 μl fluid, 10 μl fluid, 5 μl fluid, 4 μl fluid, 3 μl fluid, 2 μl fluid, 1 μl fluid, 0.5 μl A signal can be provided in response to placement of fluid, 0.3 μl fluid, 0.5 μl fluid, or 0.1 μl fluid.

  In some embodiments, the signal provider can provide a signal in response to displacement of at least one well toward the first surface.

  In certain embodiments, the at least one signal provider includes 300 milligrams of material in a well, 250 milligrams of material, 200 milligrams of material, 150 milligrams of material, 100 milligrams of material, 75 milligrams of material, 50 milligrams of material, 45 Milligram substance, 40 milligram substance, 35 milligram substance, 30 milligram substance, 25 milligram substance, 20 milligram substance, 15 milligram substance, 10 milligram substance, 5 milligram substance, 4 milligram substance, 3 A signal can be provided in response to removal of milligrams of material, 2 milligrams of material or 1 milligram of material, 500 micrograms (μg) of material, 300 μg of material, 200 μg of material or 100 μg of material.

  At least one signal provider consists of 300 microliter (μl) fluid in a well, 250 μl fluid, 200 μl fluid, 150 μl fluid, 100 μl fluid, 75 μl fluid, 50 μl fluid, 45 μl fluid, 40 μl fluid, 35 μl fluid, 30 μl fluid, 25 μl fluid, 20 μl fluid, 15 μl fluid, 10 μl fluid, 5 μl fluid, 4 μl fluid, 3 μl fluid, 2 μl fluid, 1 μl fluid, 0.5 μl fluid, 0.3 A signal can be provided in response to removal of μl fluid, 0.2 μl fluid, or 0.1 μl fluid.

  In some embodiments, at least one well in the plate is removable therefrom.

  In some embodiments, the plate comprises at least one temperature sensor associated with at least one of the wells. In some embodiments, the temperature sensor is placed on or in one of the wells. In some embodiments, the at least one temperature sensor is configured to provide a signal representative of the temperature of at least one well or its proximity. In some such embodiments, the at least one temperature sensor is configured to continuously detect the temperature of the at least one well and periodically provide a signal representative of the temperature.

  In some embodiments, the plate further comprises an electronic storage element for storage of at least one signal provided by at least one of at least one signal provider and at least one temperature sensor.

  In some embodiments, the at least one temperature sensor is configured to detect a temperature in a group of wells from a plurality of wells.

  In some embodiments, the at least one temperature sensor comprises a single temperature sensor configured to detect the temperature of all wells.

  In some embodiments, the at least one temperature sensor comprises a plurality of temperature sensors, each of the plurality of wells for detecting a temperature in one of the plurality of wells associated therewith. Related to one.

  In some embodiments, the plate further comprises at least one heating component associated with the at least one well, wherein the at least one heating component is at least one so as to heat at least one well or the interior thereof. It is installed close enough to two wells. In some embodiments, the at least one heating component comprises a plurality of heating components, each heating one well or its interior substantially without heating other portions of the plurality of wells. Therefore, it is associated with one well of the plurality of wells and is placed close enough to one well associated therewith. In some embodiments, at least one heating component comprises a heating coil. In some embodiments, the at least one heating component can also cool at least one well. In some embodiments, the heating component includes a Peltier element.

  In some embodiments, at least one well in the plate is removable therefrom in a manner that does not remove the heating component associated with the at least one removable well from the plate. In some embodiments, at least one well in the plate is removable therefrom, and the heating component associated with the at least one removable well is attached or integrally formed with the at least one well; It is removable with it.

  In some embodiments, the plate further comprises an electrical port operatively associated with at least one of the at least one signal provider and the at least one temperature sensor. In some embodiments, the plate further comprises a rechargeable power supply, wherein the rechargeable power supply is functionally associated with at least one of the at least one signal provider and the at least one temperature sensor. It is configured to be recharged by connecting it to a power generation source. In some embodiments, the rechargeable power supply is configured to be recharged when the electrical port is electrically connected to a power generation source.

  In addition, according to one embodiment of the present invention, a data reader is provided that houses a plate therein according to one embodiment of the present invention, the data reader comprising a base, an electrical And a base, the base is for mounting the plate therein, the electrical port corresponds to the electrical port of the plate for electrical engagement therewith, and the processor is electrically It is functionally associated with the electrical port to process the signal obtained from at least one of the signal provider and the temperature sensor via the port. In some embodiments, the processor is configured to obtain signals directly from at least one of a signal provider and a temperature sensor via an electrical port. In some embodiments, the processor is configured to obtain a signal from an electronic storage component that stores at least one signal provided by at least one of a signal provider and a temperature sensor.

  In some embodiments, the data reader further comprises a display functionally associated with the processor, the display configured to provide the user with information derived from the processed signal. In some embodiments, the information includes: (i) the amount of fluid in the plate at a particular time; (ii) the amount of fluid in at least one well at the particular time; and (iii) the plate over time. A change in the amount of fluid in (iv) a change in the amount of fluid in at least one well over time, (v) the temperature of at least one well at a particular time, and (vi) at least one over time. It includes at least one indication of the temperature change of one well. In some embodiments, the processor is configured to process signals in real time and the display is configured to provide real time information to the user.

  Also according to an embodiment of the present invention, a plate comprising a first substantially planar surface, a second substantially planar surface, at least one well and at least one heating component, the first substantially planar surface. Comprises at least one first opening defined therein, the second substantially planar surface is substantially parallel to the first substantially planar surface, and the second substantially planar surface is the first substantially planar surface. Spaced from the planar surface, at least one well is defined in the plate, and the at least one well corresponds to one of the at least one first opening defined in the first surface. A second opening aligned therewith, the at least one well having a sidewall and a bottom, the at least one well extending from the first substantially planar surface toward the second substantially planar surface; , At least one heating component is associated with the at least one well and is located sufficiently close to the at least one well to heat at least one well or the interior thereof. In some embodiments, the first surface comprises a plurality of first openings defined therein, at least one well comprises a plurality of wells defined in the plate, each well comprising a first A second opening corresponding to and aligned with a first opening from a plurality of first openings defined on one surface, each of the wells having a side wall and a bottom; Each of the wells has a plurality of heating elements associated therewith, extending from one substantially planar surface to a second substantially planar surface, wherein at least one heating component comprises a plurality of heating components. With one of the components, each of the heating components being placed close enough to the well with which the heating component is associated to heat the well or its interior substantially without heating the other wells The plate will be provided Is done. In some embodiments, the heating component comprises a heating coil. In some embodiments, the heating component can also cool the well. In some embodiments, the heating component includes a Peltier element. In some embodiments, at least one well in the plate is removable therefrom without removing the heating component associated with the at least one removable well from the plate. In some embodiments, at least one well of the plate is removable therefrom, and the heating component associated with the at least one removable well is attached or integrally formed with the at least one well; And it is removable with it.

  Also, according to one embodiment of the present invention, there is provided a method for measuring the amount of fluid added to a plate according to any one of the embodiments of the present invention, said method provided by a signal provider Recording an initial signal; and after adding fluid to at least one well in the plate, obtaining a second signal generated by the signal provider in response to the addition of the fluid; Based on the difference between the signals, the amount of fluid added to at least one well can be calculated. In some embodiments, the method further comprises adding fluid to the plate after the step of recording the initial signal and before the step of obtaining the second signal. In some embodiments, the method further comprises calculating the amount of fluid added to the at least one well based on the difference between the initial signal and the second signal. In certain embodiments, calculating the amount includes calculating a volume of fluid added to the at least one well. In certain embodiments, calculating the amount includes calculating a mass of fluid added to the at least one well. In certain embodiments, calculating the amount includes calculating the volume and mass of fluid added to the at least one well.

  Also, according to one embodiment of the present invention, there is provided a method for measuring the amount of fluid lost from a plate according to any one of the embodiments of the present invention, wherein the plate is placed in at least one well of the plate. Having an initial amount of fluid dispensed, wherein the method records an initial signal provided by the signal provider at a first time, and after the first time, receives a second signal from the signal provider at a second time. And obtaining an amount of fluid lost from at least one well of the plate based on the difference between the initial signal and the second signal. In certain embodiments, calculating the amount includes calculating a volume of fluid lost from at least one well. In certain embodiments, calculating the amount includes calculating the mass of fluid lost from at least one well. In certain embodiments, calculating the amount includes calculating the volume and mass of fluid lost from at least one well. In certain embodiments, the method comprises periodically repeating the step of obtaining a signal and at least one well in a period between two different times based on the difference between the signals obtained at two different times. And calculating the amount of fluid lost from.

  Also, according to one embodiment of the present invention, at least one of the wells at a time after obtaining a baseline measurement of displacement of at least one well in the multi-well plate and obtaining the baseline measurement. Obtaining a second measurement of displacement and calculating a change in the amount of fluid in at least one well based on the second measurement of displacement, wherein the multi-well plate includes at least A displacement measurement assembly for measuring the displacement of at least one well in the plate in response to a change in the amount of fluid in one well, wherein the baseline measurement is obtained via the displacement measurement assembly; Provide a method.

  Also, according to one embodiment of the present invention, obtaining a baseline measurement of the displacement of at least one well in the multi-well plate at a first time and at least one at a second time after the first time. Measuring the displacement of one well, and calculating a change in the amount of fluid in at least one well based on the change in displacement between the first and second times, the multiwell plate comprising: There is a displacement measurement assembly for measuring the displacement of at least one well in the plate in response to a change in the amount of fluid in the at least one well, and the baseline measurement is a displacement measurement assembly And at least one of the first and second times is provided, wherein a detectable amount of fluid is present in the well. . In certain embodiments, the method includes periodically repeating the step of measuring the displacement of at least one well at a second time, and based on a change in displacement between the two measurements of displacement. Calculating a change in the amount of fluid in at least one well during a period between two measurements. In certain embodiments, the change in the amount of fluid is due to the addition of fluid to at least one well. In certain embodiments, the change in the amount of fluid is due to a decrease in fluid from at least one well.

  In certain embodiments of the above method, the plate is a plate according to any one of the embodiments of the invention.

  In certain embodiments of the above methods, the signal provider is the volume of fluid in at least one well and is 300 microliters (μl), 250 μl, 200 μl, 150 μl, 100 μl, 75 μl, 50 μl, 45 μl, 40 μl, 35 μl, 30 μl, 25 μl. Sensitive enough to detect changes in 20 μl, 15 μl, 10 μl, 5 μl, 4 μl, 3 μl, 2 μl, 1 μl, 0.5 μl fluid, 0.3 μl fluid, 0.2 μl fluid or 0.1 μl fluid.

  In certain embodiments of the above methods, the signal provider has a mass of fluid in at least one well, 300 milligrams (mg), 250 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, 45 mg, 40 mg, 35 mg, 30 mg, 25 mg, Sensitive enough to detect changes of 20 mg, 15 mg, 10 mg, 5 mg, 4 mg, 3 mg, 2 mg, 1 mg, 500 micrograms (μg), 300 μg, 200 μg or 100 μg.

  In certain embodiments of the method, the method further comprises detecting the temperature of at least one well. In certain embodiments, the method further comprises detecting the temperature of at least one well at at least two different time points. In certain embodiments, the method further comprises adjusting the temperature of the individual wells in response to detecting the temperature.

  In certain embodiments of the above method, at least one well in the multi-well plate is removable therefrom.

  Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of conflict, the present specification, including definitions, will control.

  As used herein, the terms “comprising”, “including”, “having” and grammatical variants thereof specify a stated feature, integer, step or component, but 1 Nor does it exclude the addition of a plurality of additional features, integers, steps, components or groups thereof. These terms include the terms “consisting of” and “consisting essentially of”.

  As used herein, the indefinite articles “a” and “an” mean “at least one” or “one or more” unless the context clearly dictates otherwise.

  Embodiments of the methods and / or devices of the present invention may include performing or completing selected tasks manually, automatically, or a combination thereof. Certain embodiments of the invention are implemented using components that include hardware, software, firmware, or a combination thereof. In some embodiments, one component is a general purpose component (eg, a general purpose computer or monitor). In some embodiments, a component is a dedicated or custom-made component (eg, a circuit, integrated circuit, or software).

  For example, in some embodiments, part of one embodiment is implemented as multiple software instructions executed by a data processor, eg, part of a general purpose or custom made computer. In some embodiments, the data processor or computer includes a volatile memory for storing instructions and / or data and / or a non-volatile storage device for storing instructions and / or data (e.g., magnetic hard disks and (Or removable media). In some embodiments, the implementation includes a network connection. In some embodiments, implementations include a user interface that typically includes one or more input devices (e.g., that allows input of commands and / or parameters), and output devices (e.g., results and operations). Which allows reporting of the parameters).

BRIEF DESCRIPTION OF THE DRAWINGS Certain embodiments of the present invention are described herein with reference to the accompanying figures. The description, together with the figures, makes it clear to those skilled in the art how certain embodiments of the invention can be implemented. The figure is for illustrative purposes only and does not attempt to show the structural details of one more detailed embodiment other than that required for a basic understanding of the present invention. . For clarity, some objects represented in the figures may not be in a fixed ratio.

FIG. 1 is a perspective view of a prior art multi-well plate. FIG. 2A is a perspective view of a multiwell plate constructed and operative in accordance with one embodiment of the present invention. FIG. 2B is an exploded view of the multiwell plate of FIG. 2A. 2C is a perspective cross-sectional view of the multi-well plate of FIGS. 2A and 2B, taken along section line IIC-IIC of FIG. 2A. 2D is a perspective cross-sectional view of the multi-well plate of FIGS. 2A and 2B along the cross-sectional line IID-IID of FIG. 2A. FIG. 3A is a perspective view of a multiwell plate constructed and operative in accordance with another embodiment of the present invention. FIG. 3B is a perspective view of a multiwell plate constructed and operative in accordance with another embodiment of the present invention. FIG. 3C is an exploded view of the multiwell plate of FIGS. 3A and 3B. FIG. 3D is an enlarged perspective view of the support, arms and blocks that form part of the multi-well plate of FIGS. 3A and 3B. 3E is a cross-sectional view of the multi-well plate of FIGS. 3A-3C taken along section line IIIE-IIIE of FIG. 3A. 3F is a cross-sectional view of the multiwell plate of FIGS. 3A-3C taken along section line IIIF-IIIF of FIG. 3A. 3G is a cross-sectional view of the multi-well plate of FIGS. 3A-3C taken along section line IIIG-IIIG of FIG. 3A. FIG. 4A is a perspective view of a multiwell plate constructed and operative in accordance with yet another embodiment of the present invention. FIG. 4B is an exploded view of the multiwell plate of FIG. 4A. 4C is a cross-sectional view of the multi-well plate of FIGS. 4A and 4B taken along section line IVC-IVC of FIG. 4A. FIG. 5A is a screenshot illustrating a graphical user interface for additional online (real-time) monitoring of fluid to a multi-well plate, in accordance with embodiments taught herein. FIG. 5B is a screenshot illustrating a graphical user interface for additional online (real-time) monitoring of fluid to a multi-well plate in accordance with embodiments taught herein. FIG. 5C is a screenshot illustrating a graphical user interface for additional online (real-time) monitoring of fluid to a multi-well plate, according to embodiments taught herein. FIG. 5D is a screenshot illustrating a graphical user interface for additional online (real-time) monitoring of fluid to a multi-well plate, in accordance with embodiments taught herein. FIG. 6A is a screenshot illustrating a graphical user interface for off-line volume monitoring of fluid in a multi-well plate, in accordance with embodiments taught herein. FIG. 6B is a screenshot illustrating a graphical user interface for off-line volume monitoring of fluid in a multi-well plate, in accordance with embodiments taught herein. FIG. 7 is a screenshot illustrating a graphical user interface for offline temperature monitoring of fluids in a multi-well plate in accordance with an embodiment of the present invention. FIG. 8 is a perspective view of a data reader and plate base constructed and operative in accordance with one embodiment taught herein to accept signals from a multiwell plate in accordance with the teachings herein. FIG. 9A is for removing a well-providing element or well-defining element from a device constructed and operative in accordance with one embodiment taught herein, and / or for placing that element in a multi-well plate. It is a perspective view of a device. FIG. 9B is for removing a well-providing element or well-defining element from a device constructed and operative in accordance with one embodiment taught herein, and / or to place such device in a multi-well plate. It is a perspective view of a device.

DESCRIPTION OF SOME EMBODIMENTS OF THE INVENTION The principles, applications and implementations taught herein may be better understood with reference to the accompanying description and figures. By reading the description and figures of the present specification, those skilled in the art can implement the present invention without undue trial and error.

  Before describing at least one embodiment of the present invention in detail, the present invention, in its application, is described in the following description and / or the arrangement of components and / or methods shown in the drawings and / or examples and It should be understood that the details are not limited to structural details. The invention can be implemented with other embodiments and can be implemented or carried out in various ways. It is also well understood that the wording and terminology used herein is for descriptive purposes and should not be taken as a limitation.

  Reference is now made to FIG. 1, which is a perspective view of a prior art multi-well plate. FIG. 1 shows a well-known typical 96-well plate 10. The plate 10 includes an upper surface 12, a lower surface 14, and a plurality of side surfaces 16 between the upper surface 12 and the lower surface 14. A plurality of wells 17 extend between the surfaces 12 and 14, and a total of 96 wells are arranged in 8 rows, each with 12 wells. Each well has an opening 18 formed in the upper surface 12 to facilitate injection of sample fluid into the well. The plate 10 may be made of plastic (eg, polystyrene or polycarbonate or a combination of such materials) and in some cases may have a glass bottom. In general, the lower surface 14 is transparent at least in a certain light frequency or range of light frequencies where a sample placed in the well is observed. However, for certain applications, such transparency may not be essential. Side 20 of well 17 may or may not be transparent depending on the type of observation being performed and the nature of the sample.

  Wells are typically circular cross-sections in the xy plane and are essentially cylindrical in shape, but may be cross-sections in other shapes (e.g., rectangular or square) along their length For example, the side surface of the well may be tapered from the top opening to the bottom of the well along a part or the entire length thereof. The bottom of the well is generally flat, but depending on the intended use of the plate, other shapes (e.g., cone, frustoconical or spherical bottom) (i.e., when viewed along the z-axis, the bottom is V-shaped or It may be formed in a U-shaped cross section.

  Plates such as plate 10 are typically 85-86 mm wide and 127-128 mm long, differing in overall height in the range 10-20 mm, and the center of the well is 9 mm along the x and y axes Are placed at intervals. Standards for the 96-well microplate footprint and bottom outer flange are described in ANSI / SBS 1-2004 and ANSI / SBS 3-2004, respectively. That said, the plates can have various length, width and height dimensions, and can have various distances between wells, so that they can be suitable for the particular application or use of the plate. Can have various numbers of wells. For example, the plate is a 384 well plate with 384 wells each having 24 wells arranged in 16 rows (so that the centers of the wells are 4.5 mm apart along the x and y axes). May be. Standards for 384-well microplate footprint and bottom outer flange are described in ANSI / SBS 1-2004 and ANSI / SBS 3-2004, respectively.

  Here, FIG. 2A is a perspective view of a multi-well plate 100 constructed and operative in accordance with an embodiment of the present invention, FIG. 2B is an exploded view of the multi-well plate 100 of FIG. 2A, and section line IIC- Reference is made to FIGS. 2C and 2D, which are perspective cross-sectional views of the multiwell plate 100 of FIGS. 2A and 2B, along IIC and IID-IID, respectively.

  As shown in FIG. 2A, because the plate 100 is designed for use with existing equipment, it is the standard plate size dimension currently used, and its wells are ANSI / SBS1 -Spaced as specified in -2004 and ANSI / SBS3-2004. Thus, when assembled, the plate 100 comprises an upper surface 112, a lower surface 114 and a plurality of side surfaces 116 between the surfaces 112 and 114, and looks similar to a typical 96 well plate. A plurality of wells 117 are formed in the plate 100 and extend between the surfaces 112 and 114. Each well includes an opening 118 formed in the upper surface 112 to facilitate injecting sample fluid into the well.

  Turning to FIG. 2B, which is an exploded view of the plate 100, it can be seen that the plate 100 is actually made up of several parts. The side surface 116 forms a part of the frame 122. The frame 122 forms a pair of supports 124a and 124b on its inner portion at each longitudinal end of the frame. The support is described in further detail in FIGS. 2C and 2D. Each of the supports 124a and 124b has attached to it a pair of flexible arms 126 consisting of an upper arm 126a and a lower arm 126b, each arm being attached to one of the supports at its proximal end. At its distal end, it is attached to block 128. As seen in the enlarged portion of FIG. 2B, each block 128 includes an upper portion and a lower portion, and arms 126a and 126b are attached to the lower surfaces 128a and 128b of the upper and lower portions, respectively. The flexible arm 126 may be made of a material that is generally flexible and generally metal or plastic capable of sensing the addition of a few micrograms of weight to the well, as will be described in more detail later. Good. The arm may be attached to the block 128 by suitable means (eg, gluing, possibly melting or welding).

  The cylindrical side wall of the well 117 is formed in the well-providing element 130, which may be formed of plastic, glass or other suitable material. The well-providing element 130 comprises an upper surface 112 and a flange 132 at its longitudinal end. When the plate 100 is assembled, each flange 132 rests on the upper surface of the upper portion 128a of the two blocks 128 (one block at each end of the flange) and is optionally attached thereto In some embodiments, it rests on supports 124a and 124b (see FIG. 2D). The bottom piece 134 is a piece of plastic or glass approximately 170-1000 microns in thickness, in a manner that seals at the end of each well to form the bottom of each well 117 below the well-providing element 130. Attached in a sealing manner.

  In the embodiment shown in FIGS. 2A-2D, a flange 132 is provided around the uppermost portion of the well-providing element 130 and is slightly smaller than the inner periphery of the frame 122. As a result, when the flange 132 of the well-providing element 130 rests on the block 128, there is a small gap 138 between the well-providing element 130 and the frame 122 (see FIG. 2D). As will be described below, the presence of the gap 138 allows movement of the well-providing element 130 along the vertical axis and concomitant deformation of the arm 126. To seal the gap 138 without hindering such movement, a very thin (approximately 7 micron) flexible material (e.g., suitable plastic) film 140 is applied to the top edge 142 of the frame 122 and the well Attaching to the outer edge of the top surface 112 of the included element 130. One way to achieve such attachment to the well-providing element 130 is by including a slightly raised rim 143 around the upper edge of the element 130. As a result, the film 140 is positioned over the gap 138 when engaged with the rim 143 and attached to the outer edge of the top surface 112.

  As shown in FIGS. 2A and 2B, the film 140 is hollow in the center so that most of the upper surface 112, particularly the region of the surface 112 comprising the well 117, is exposed and the film 140 is It only covers the edges of the included element 130, the gap 138 and the upper edge 142 of the frame 122. However, in some embodiments, if a plurality of circular openings are formed in the film 140 that are aligned with the openings 118 of the well-providing element 130 and the user has access to the well 117, the film 140 may be Needless to say, the entire upper surface 112 and the upper edge 142 of the frame 122 may be covered. In such embodiments, the well-providing element 130 may be formed with a slightly raised portion or rim around the opening 118 to provide an additional surface to which the film 140 can be secured.

  Film 140, well-providing element 130, bottom piece 134 and frame 122 are attached to each other using any suitable means (eg, soldering, gluing, fusing, bonding or any other suitable attachment mechanism).

  As described above, when the plate 100 is assembled, the well-providing element 130 rests on a pair of blocks 128 at each longitudinal end thereof and is optionally attached thereto. This limits the movement of element 130 in the x- and y- directions, but movement in the vertical (z) direction is possible. Thus, when fluid enters the well 117 of the element 130, the weight of the fluid causes a downward displacement of the element 130 and a deformation of the arm 126. In general, it is needless to say that only a few microliters of fluid is added to the predetermined well 117 depending on the purpose of use. As a result, the flexible arm 126 must be adequately flexible to bend in response to the addition of a few micrograms of fluid to the well.

  Further reference is made to FIGS. 2C and 2D which show the support 124a and 124b, the arm 126, the block 128 and the attachment of the arm 126 to the support in more detail. FIG. 2C shows a plate end of the plate 100 of FIG. 2A in a perspective cross-sectional view (cross-sectional view along the cross-sectional line IIC-IIC in FIG. 2A). FIG. 2D shows an enlarged cross-sectional view of the same plate at the same angle as FIG. 2C along the cross-sectional line IID-IID in FIG. 2A, which shows the other end of the plate. FIG. 2C shows the pair of arms 126 closest to the plate end, while FIG. 2D shows the pair of arms 126 positioned slightly inward from the plate end.

  The support 124a protrudes from both the inner wall 122a installed at one of the longitudinal ends of the frame 122 and the longitudinal wall 122b adjacent thereto. The support 124b protrudes from the opposite longitudinal wall 122c and is spaced from the inner wall 122a by the width of the support 124a. The supports 124a and 124b protrude in opposite directions from the longitudinal wall. Each of the supports 124a and 124b protrudes about 1/3 between the longitudinal walls, and each support further protrudes approximately halfway between the two longitudinal walls (see FIG. 2D includes support 124b's 124b 'and 124b' '). The distance between the upper surfaces of the tongues is equal to the distance between the arms 126a and 126b. A pair of arms is attached at its proximal end to support the tongues 124a 'and 124a' '(not shown) of the support 124a, and the other pair of arms is at its proximal end. It is attached to the tongues 124b 'and 124b' 'of the support 124b. Each pair of arms is attached to block 128 at its distal end. As described above, the element 130 is placed so as to rest on and be attached to the block 128. If this position is considered to be a neutral or basic position, the arm 126 will be flexible enough to deform from this neutral position in the z-axis direction as liquid is added to several wells 117. Configured to have.

  Thin and flat strain gauges 144 are attached to the upper and lower surfaces of each arm 126 in the free region of the arms, but in some embodiments are near the point where the arms are attached to the frame 122. Each of the strain gauges is electrically connected (eg, by a thin wire (not shown)) to a thin and flat electronic card 146 installed under the lower arm 126b. Preferably, the electronic card 146 is arranged to minimize the connection length between the strain gauge 144 and the card 146. Card 146 is electrically coupled to a processor (not shown). It goes without saying that the use of the strain gauge 144 and the card 146 enables a correlation between a change in the electrical resistance of a circuit installed in the card 146 (for example, measurement using a Wheatstone bridge) and the bending of the arm 126. . This allows the calculation of the mass of fluid added to the well. Thus, arm 126 with strain gauge 144 forms a signal provider that provides a signal that indicates a change in the amount of fluid in one or more wells. If the density of the fluid is known, calculation of the volume of fluid added is facilitated. In some embodiments, the plate 100 further comprises a power supply (eg, a storage battery) (not shown) that connects to the electronic card 146, for example, and the power supply supplies power to the electronic components of the plate 100. If the plate 100 is connected to a power source or computing device via an appropriate port (not shown), it can be recharged.

  For example, if rows A through L of plate 100 are filled sequentially using an 8-tip pipette, it is possible to repeatedly calculate the amount of fluid added to each row, so that the wrong amount of fluid is Can be specified for each row in real time. This allows extraordinary rows to be removed from the calculation at the end of the experiment rather than discarding the results for all plates. Preferably, the device used to add fluid to the well is equipped with control software that allows only the affected rows to be excluded from further manipulation during the remainder of the experiment (eg addition of reagents and reactants). Even if all wells are filled at the same time, the use of plate 100 in conjunction with appropriate software that identifies the addition of the wrong amount of fluid to the plate in real time will allow the user to use that abnormal plate. It goes without saying that it is possible to avoid waste of reagents and reactants in downstream experiments by making it possible to stop ongoing experiments.

  According to an embodiment of the invention, the plate 100 comprises means for heating and optionally cooling individual wells. Such heating means may be, for example, in the form of (a) a heating coil placed around at least some of the wells, or (b) a Peltier element (sometimes called a Peltier heat pump or thermoelectric cooler). Good. As is apparent, Peltier elements can be used to cool as well as heat individual wells, and can also detect or monitor the temperature of the wells or their surroundings. In this way, the temperature of individual wells can be controlled. For example, the temperature of each well can be maintained at 37 ° C. ± 0.5 ° C., respectively.

  In one embodiment, a single heating coil or Peltier element may be placed on the well-providing element 130 to heat two or more wells 117 together. In another embodiment, multiple heating coils or Peltier elements may be placed in one or more individual wells 117 for heating them.

  If the temperature of the well is measured periodically, and if the device used to measure the temperature is connected to a controller that controls the heating means for each individual well, then the temperature of the individual well is measured periodically It goes without saying that the individual well heating means used in conjunction with can provide a method for improving control over the conditions of a given well. Thus, for example, the plate 100 having such heating means is stored in a thermostat, the temperature of the well is periodically monitored, and the temperature of the individual well is heated (or cooled) as necessary. You may adjust by doing. Alternatively, the heating means itself may be used for incubation, e.g., with frequent temperature monitoring and adjustment (i.e., every 15, 10 or 5 minutes, for example) to specify that incubation is to occur. The well temperature may be maintained at 37 ° C. ± 0.5 ° C., for example. Thus, by providing individual heating elements for each well in plate 100, well-providing element 130 or well 117, the temperature can be adjusted and controlled if the temperature is found to be incorrect.

  Reagents and other fluids can be removed from the plate 100, added to or removed from the well 117 in the well-providing element 130, which can be optionally discarded, so that during each use of the plate 100 It will be appreciated that the plate 100 may be used multiple times and / or in multiple experiments if the included element 130 can be replaced.

  Reference is now made to FIGS. 3A-3G illustrating a multi-well plate 300 constructed and operative in accordance with an embodiment of the present invention. 3A and 3B are perspective views of the multiwell plate 300, FIG.3C is an exploded view of the multiwell plate 300, and FIG.3D forms part of the multiwell plate 300. FIGS. 3E, 3F and 3G are cross-sectional views of the multiwell plate 300 along cross-sectional lines IIIE-IIIE, IIIF-IIIF and IIIG-IIIG of FIG. 3A. .

  As shown in FIGS. 3A and 3B, because the plate 300 is designed for use with existing equipment, it is dimensioned according to the standard plate size currently used, and its wells are similarly There is a gap. Thus, when assembled, plate 300 comprises a plurality of wells 317 formed therein, with a plurality of wells 317 formed thereon, with an upper surface 312, a lower surface 314, a plurality of side surfaces 316 between surfaces 312 and 314, and is similar to a typical 96 well plate. Looks.

  As shown in FIG. 3C, which is an exploded view of the plate 300, the plate 300 is actually formed of several parts. The side surface 316 forms a part of the frame 322. The frame 322 forms a pair of supports 324a and 324b on its inner part at the longitudinal end of the frame. Each support includes a plurality of “stairs” 325. The support is described in further detail in FIGS. 3D, 3F and 3G. The frame 322 includes end walls 322a and 322d and longitudinal walls 322b and 322c disposed at the longitudinal ends of the frame.

  Each of the supports 324a and 324b has a plurality of flexible arms 326 attached to it, each arm being attached to one of the supports at its proximal end and to the block 328 at its distal end. It is attached. The structure and function of flexible arm 326 and block 328 are described in more detail below with respect to FIGS. 3D, 3F, and 3G. The flexible arm 326 is a suitable material, typically a metal or plastic that is suitably flexible and capable of sensing the addition or removal of small volumes to or from the well, as will be described in more detail later. It may be made. For example, flexible arm 326 is less than 300 microliters (μl), less than 250 μl, less than 200 μl, less than 150 μl, less than 100 μl, less than 75 μl, less than 50 μl, less than 45 μl, less than 40 μl, less than 35 μl, less than 30 μl, less than 25 μl, Addition to wells <20 μl, <15 μl, <10 μl, <5 μl, <4 μl, <3 μl, <2 μl or <1 μl could be detected.

  The arm 326 may be attached to the block 328 by suitable means (adhesion, possibly melting or welding).

  The plate 300 further comprises eight identical well support elements 330, each with a longitudinal opening 333 suitable for receiving a well-providing element 348 therein. Each well-providing element 348 comprises twelve cylinders 320 and forms the side wall of a cylindrical well 317 formed therein. Each of the elements 330 also includes a pair of flanges 332 with one flange at each of its longitudinal ends. Well support element 330 may be formed of plastic or other suitable material, and may include other materials (eg, glass). When assembled, each flange 332 rests on and preferably is attached to two blocks 328 (one block per flange). The manner in which block 328 is held in place relative to frame 322 is described in further detail below.

  The plurality of bottom pieces 334 are plastic or glass pieces approximately 170-1000 microns thick and sealed at the well ends of each well to form the bottom of each well 117 below the well-providing element 348. In a sealing manner. In the illustrated embodiment, the three pieces 334 each seal four wells 317 and are attached to each well-providing element 348. That said, the bottom piece may use any suitable arrangement or number, for example, all wells in a single well comprising element 348 may be sealed with a single piece 334. Each well 317 may be sealed with an individual bottom piece 334, and six bottom pieces 334 may be used to seal a pair of wells 317 in a single well-providing element 348. In some embodiments, the frame 322 includes a divider 350 that divides the frame into sections 352 adjacent to the bottom portion thereof, each section 352 including one of the bottom pieces 334 and wells attached thereto. 317 is set with appropriate dimensions to fit.

  When assembled together, the periphery of the top portion of the collection of well support elements 330 is provided with a flange 332, which is slightly smaller than the inner periphery of the frame 322, resulting in the flange 332 of the well support element 330. Of course, there is a small gap 338 between the element 330 itself (see FIG. 3E) and between the well support element 330 and the frame 322 (see FIG. 3E). . As described below, the presence of the gap 338 allows movement of the well support element 330 along the vertical axis and concomitant deformation of the arm 326. In order to seal the gap 338 without hindering such movement, a very thin (e.g., approximately 7 microns) flexible material (e.g., suitable plastic) film 340 is applied to the upper edge 342 of the frame 322, and And attach to at least a portion of the upper surface of the well support element 330. The film 340 includes eight rectangular openings 343 that are aligned with the openings 333 of the assembled well support element 330.

  Film 340, well support element 330, bottom piece 334, and frame 322 are attached to each other using any suitable means (eg, soldering, gluing, melting, bonding, or any other suitable attachment mechanism). In the illustrated embodiment, the well-providing element 348 can be easily removed from the well support element 330 without damaging the structure and function of the sensitive arm 326 (detailed below). To state).

  As described above, when the plate 300 is assembled, the well support elements 330 rest on and attach to the pair of blocks 228 at their longitudinal ends. This limits the movement of element 330 in the horizontal and vertical (x and y) directions, but movement in the vertical (z-) direction is possible. Thus, when fluid enters (or is removed from) a well of a well-providing element 348 installed on any of the well support elements 330, an increase (or decrease) in the weight of the fluid causes the well support element 330 to Deformation of the attached arm 326 is caused. In general, it will be appreciated that only a few microliters or less of fluid is added to a given well 317, depending on the intended use, up to several hundred microliters. As a result, the flexible arm 326 must be properly flexible to bend in response to the addition (or removal) of such an amount (or equivalent mass) of fluid to (or from) the well. For example, less than 300 microliters (μl), less than 250 μl, less than 200 μl, less than 15 μl, less than 100 μl, less than 75 μl, less than 50 μl, less than 45 μl, less than 40 μl, less than 35 μl, less than 30 μl, less than 25 μl, less than 20 μl, less than 15 μl, 10 μl Less than, less than 5 μl, less than 4 μl, less than 3 μl, less than 2 μl, or less than 1 μl of fluid may be added to the well 317 at a given time.

  A thin and flat strain gauge 344 attaches to the upper and lower surfaces of each arm 326 near the point where the arm is attached to one of the “stairs” 325 (which will be described in more detail below). Rather than placing strain gauges on the upper and lower surfaces of the arm, instead, there are two strain gauges, one on the upper surface of the arm, in the vicinity where the arm is attached to the frame 322, and the arm is block 328. Another one may be placed near where it is attached, one with two strain gauges, one on the lower side of the arm, near where the arm is attached to the frame 322, and the arm attached to the block 328 It goes without saying that only one more may be placed in the vicinity.

  Each strain gauge 344 is electrically connected (eg, by a thin wire (not shown)) to a thin, flat electronic card 346 that is placed under the lower arm 326. Preferably, the electronic card 346 is arranged to minimize the connection length between the strain gauge 344 and the card. The card 346 is electrically coupled to a port 358 installed in the frame 322 via a plurality of wires 356. As described below with respect to FIGS. 8-9B, the ports are configured for connection to appropriately equipped plate bases and data reader electrical ports. In some embodiments, the data reader may execute a graphical user interface described below with respect to FIGS. Preferably, the port 358 does not extend beyond the outer surface of the frame 322, and is preferably flush with the outer surface and does not affect the overall dimensions of the plate 300, Can be used. In some embodiments, the plate 300 further comprises a power supply (eg, a storage battery) (not shown) that connects to the electronic card 346, for example, which provides power to the electronic components of the plate 300. If the plate 300 is connected to a power generation source or computer device via the port 358 or USB port, it can be recharged.

  The electronic card 346 may have elements thereon for measuring the electrical resistance of the circuit (eg, Wheatstone bridge). Bending of arm 326 caused by a change in volume or weight contained in one or more wells 317 causes a change in the length of the corresponding strain gauge 344 resistor. This change in the strain gauge 344 correlates with a change in the electrical resistance of the circuit. This change is measured at an element on the electronic card 346 using, for example, a Wheatstone bridge. Changes in the electrical resistance of the circuit allow calculation of the mass of fluid added (or removed) to the well 317. Specifically, the greater the mass of fluid added to (or removed from) the well, the greater the change in flex of arm 326 and the greater the change in electrical resistance of the circuit. Therefore, the measured value of the change in the electrical resistance of the circuit indicates the change in the mass of the fluid in the well, and the change can be calculated based on the measured value. If the density of the fluid is known, calculation of the volume of fluid added (or removed) is facilitated. Thus, arm 326 with strain gauge 344 forms a signal provider that provides a signal that indicates a change in the amount of fluid in one or more wells.

  For example, if rows A through L of plate 300 are filled sequentially using an 8-tip dispenser, it is possible to repeatedly calculate the amount of fluid added to each well in each row, so the wrong amount Whether too much or too little fluid is added to a particular row, it can be identified column by column in real time. Preferably, the device used to add fluid to the well comprises control software that the device can use to correct errors. In cases where too little fluid is added, additional fluid may be distributed to the affected wells so that the fluid in the well reaches the appropriate amount and / or the software may It may be possible to adjust the error by adding a reagent or reaction solution that is insufficient to the affected well. Similarly, if too much fluid is added to a particular well, it may be appropriately scaled up by adding reagents or reactants in further operations.

  Alternatively, the affected wells or well components may be included in further manipulations for the remainder of the experiment, and the results for a particular well will calculate the wrong volume used during the experiment. It may be used in the calculation after the end of the experiment by adjusting and eliminating.

  Yet another method is to identify abnormal wells with the wrong amount of fluid added to the wells, rather than discarding the results for all rows or columns or plates where the wells are located, after the end of the experiment. The abnormal well can be excluded from the calculation.

  Even if all wells are filled at the same time, the use of plate 300 in conjunction with the appropriate software to identify the addition of the wrong amount of fluid to a particular well support element 330 in real time will allow the user to Of course, it is possible to avoid wasting reagents and reactants in downstream experiments by allowing the ongoing experiment using wells 330 or plate 300 wells to be stopped. .

  In addition, the electronic card 346 has components for manipulating data collected by various sensor elements connected to the card (e.g., an analog-to-digital conversion configuration for converting Wheatstone bridge analog signals to digital signals). The elements and normalization components for normalizing the collected signal) may be electrically coupled.

  In some embodiments, the plate 300 is electrically coupled to the electronic card 346 near one or more wells 317 and is configured to provide an indication of temperature or temperature change. A temperature sensor (not shown) is also provided. Because temperature changes in the system can affect the strain gauge 344, knowledge of changes to temperature and computational considerations allow for more accurate identification of well weights and stabilization of well sample temperature. It goes without saying that it will be possible to ensure reliability and increase sensitivity to temperature changes.

  In accordance with an embodiment of the invention, the plate 300 comprises means for heating, optionally cooling individual wells. Such heating means may be, for example, in the form of (a) a heating coil placed around at least some of the wells, or (b) a Peltier element (sometimes called a Peltier heat pump or thermoelectric cooler). Good. As is apparent, Peltier elements can be used to cool as well as heat individual wells, and can also detect or monitor the temperature of the wells or their surroundings. In this way, the temperature of individual wells can be controlled. For example, the temperature of each well can be maintained at 37 ° C. ± 0.5 ° C., respectively.

  In some embodiments, a heating coil or Peltier element may be placed on part or all of the well support element 330 to collectively heat two or more wells supported by the well support element. . In other embodiments, heating coils or Peltier elements may be placed on some or all of the individual wells 317 (e.g., their cylinders) such that each heating coil or Peltier element heats a particular well 317 on which it is placed. 320) may be arranged.

  If the temperature of the well is measured periodically, and if the device used to measure the temperature is connected to a controller that controls the heating means for each individual well, then the temperature of the individual well is measured periodically It goes without saying that the individual well heating means used in conjunction with can provide a method for improving control over the conditions of a given well. Thus, for example, the plate 300 having such heating means is stored in a thermostat, the temperature of the well is periodically monitored, and the temperature of the individual well is heated (or cooled) as necessary. You may adjust by doing. Alternatively, the heating means itself may be used for incubation, e.g., with frequent temperature monitoring and adjustment (i.e., every 15, 10 or 5 minutes, for example) to specify that incubation is to occur. The well temperature may be maintained at 37 ° C. ± 0.5 ° C., for example. Thus, by providing individual heating elements for each well in plate 300, well support element 330 or well 317, the temperature can be adjusted and controlled if the temperature is found to be incorrect.

  Here, FIG. 3D is an enlarged perspective view of the supports 324a and 324b including the arm 326 and the block 328, and FIGS. 3F and 3F are sectional views taken along the sectional lines IIIF-IIIF and IIIG-IIIG of FIG. 3A, respectively. Refer to 3G. Figures 3D, 3F and 3G show in more detail the supports 324a and 324b, the arm 326, the block 328 and the attachment of the arm 326 to the support.

  As shown, eight blocks 328 are placed near each longitudinal end of frame 322 so that each well support element 330 has two blocks 328 (one at each longitudinal end of element 330). ) And mounted. To accommodate eight blocks at each end of frame 322, supports 324a and 324b and block 328 are designed in a staircase fashion (described below). The first support 324a protrudes from one longitudinal end inner wall 322d of the frame 322 and a longitudinal wall 322b adjacent thereto. At the other longitudinal end of the frame 322, the second support 324a protrudes from the longitudinal wall 322b but is slightly spaced from the inner wall 322a. In both cases, to accommodate the four pairs of arms 326 and to facilitate proper positioning of the block 328, the support 324a is formed of a lower set 360b, an upper set 360a and two sets of parallel formed. It consists of a staircase style with “staircase”. All of the “stairs” of the support 324a protrude from the inner wall 322b of the frame 322, but all of the “stairs” have the same width along the longitudinal axis of the frame 322, and each set of uppermost “stairs” 325a and 325a ′ protrude the smallest from the inner wall 322b, “stairs” 325b and 325b ′ protrude slightly, “stairs” 325c and 325c ′ protrude more, and “stairs” 325d and 325d ′ protrudes in the middle between the wall 322b and the wall 322c.

  The distance between the upper surfaces of each pair of corresponding "staircases" (i.e., between 325a and 325a ', between 325b and 325b', between 325c and 325c ', and between 325d and 325d') Is the same as the distance between the lower surface of each pair. Thus, the first pair of arms 326a and 326a ′ is attached at its proximal end to the steps 325a and 325a ′, and the second pair of arms 326b and 326b ′ is at its proximal end, steps 325b and 325b ′. A third pair of arms 326c and 326c ′ is attached at its proximal end to stairs 325c and 325c ′, and a fourth pair of arms 326d and 326d ′ is at its proximal end, stairs 325d and Attach to 325d '. “Stairs” are spaced from each other so that the arms in each pair of arms are parallel to each other, so that in normal use, each pair of arms does not contact the other pair of arms. Can move on.

  As shown in FIGS. 3D and 3F, at their distal ends, arms 326 are attached to block 328 such that each pair of arms is attached parallel to a single block 328. To facilitate the compact placement of the arms, each block 328 includes an upper inner surface (for attaching the first arm thereto) and a lower inner surface (for attaching the second arm thereto). , A recess (which allows an additional arm to pass through). Accordingly, the arm 326a attached to the staircase 325a is attached to the upper inner surface 328a ′ of the block 328a closest to the center of the frame, and the block 328a is configured with a recess 328a ″. This is intended to allow arms 326b, 326c and 326d to pass therethrough without touching each other and without touching 328a. Similarly, arm 326a ′ attached to staircase 325a ′ is attached at its distal end to the bottom surface 328a ′ ″ of block 328a, and arms 326b ′, 326c ′ and 326d ′ Pass under. Similarly, block 328b allows attachment of arms 326b and 326b 'at surfaces 328b' and 328b '' 'respectively, passing therethrough of arms 326c and 326d and passing below arms 326c' and 326d '. Is configured to be possible. Block 328c is configured to allow attachment of arms 326c and 326c ′ at surfaces 328c ′ and 328c ′ ″, allowing passage therethrough of arm 326d and passage thereunder of arm 326d ′. Of the four blocks 328a, 328b, 328c and 328d, the block 328d closest to the wall 322c is configured with surfaces 328d 'and 328d' '' to allow attachment of the arms 326d and 326d ', respectively. .

  As a result of this configuration, it goes without saying that the sizes of the upper portions of the blocks 328a, 328b, 328c and 328d are different from each other. Each of the blocks 328a, 328b, 328c and 328d is different in overall height and the position of the top surface of each block is preferably the same with respect to the top of the frame 322, while the top surface of each block Of course, the position can be slightly different, and by using software that calculates the displacement, the mass of fluid added to the well can be programmed to eliminate such differences. For the same reason, it is not essential that all blocks 328 have the same mass.

  As clearly shown in FIG. 3G, the supports 324b are arranged in a similar manner that is in the opposite direction to the support 324a. At one longitudinal end of the frame 322, the first support 324b protrudes from an inner wall 322a at one longitudinal end of the frame 322 and a longitudinal wall 322c adjacent thereto. At the other longitudinal end of the frame 322, the second support 324b protrudes from the longitudinal wall 322c, but is slightly spaced from the inner wall 322d. Near wall 322d, support 324b is spaced therefrom to accommodate arms 326a, 326a ′, 326b, 326b ′, 326c, 326c ′, 326d, and 326d ′. To accommodate the four pairs of arms 326 and facilitate proper positioning of the block 328, the support 324b comprises a lower set 362b, an upper set 362a and two sets of “stairs” formed in parallel. It is composed in a staircase style. All of the “staircase” of the support 324b protrudes from the inner walls 322a (or 322d) and 322c of the frame 322, but all of the “staircase” has the same width, so from the wall 322a (or 322d) Projecting the same distance, each set 325e and 325e ′ of the top “staircase” projects the smallest from the inner wall 322c, “staircases” 325f and 325f ′ project slightly more, and “staircases” 325g and 325g ′ The “stairs” 325h and 325h ′ protrude in the middle between the walls 322b and 322c.

  The distance between the upper surfaces of each pair of corresponding "staircases" (i.e., between 325e and 325e ', between 325f and 325f', between 325g and 325g ', and between 325h and 325h') Is the same as the distance between the lower surface of each pair. Thus, the first pair of arms 326e and 326e ′ is attached at its proximal end to the stairs 325e and 325e ′, and the second pair of arms 326f and 326f ′ is at its proximal end, the stairs 325f and 325f ′. A third pair of arms 326g and 326g ′ is attached at its proximal end to stairs 325g and 325g ′, and a fourth pair of arms 326h and 326h ′ is at its proximal end, stairs 325h and Attach to 325h '. “Stairs” are spaced from each other so that the arms in each pair of arms are parallel to each other, so that in normal use, each pair of arms does not contact the other pair of arms. Can move on.

  As shown in FIGS. 3D and 3G, at their distal ends, arms 326 are attached to block 328 and each pair of arms is attached in parallel to a single block 328. To facilitate the compact placement of the arms, each block 328 includes an upper inner surface (for attaching the first arm thereto) and a lower inner surface (for attaching the second arm thereto). , A recess (which allows an additional arm to pass through). Accordingly, the arm 326e attached to the staircase 325e is attached to the upper inner surface 328e ′ of the block 328e closest to the center of the frame, and the block 328e is configured with a recess 328e ″. This is intended to allow arms 326f, 326g and 326h to pass therethrough without touching each other and without touching 328e. Similarly, arm 326e 'attached to staircase 325e' is attached at its distal end to the bottom surface 328e '' 'of block 328e, and arms 326f', 326g 'and 326h' Pass under. Similarly, block 328f allows attachment of arms 326f and 326f 'at surfaces 328f' and 328f '' ', respectively, passing there through arms 326g and 326h, and passing below arms 326g' and 326h '. Is configured to be possible. Block 328g is configured to allow attachment of arms 326g and 326g ′ at surfaces 328g ′ and 328g ′ ″, allowing passage therethrough of arm 326h and passage beneath arm 326h ′. Of the four blocks 328e, 328f, 328f and 328g, the block 328h closest to the wall 322b is configured with surfaces 328h 'and 328h' '' to allow attachment of arms 326h and 326h ', respectively. .

  As a result of this configuration, it goes without saying that the sizes of the upper portions of the blocks 328e, 328f, 328g and 328h are different from each other. Each of the blocks 328e, 328f, 328g, and 328h are different in overall height and the position of the top surface of each block is preferably the same with respect to the top of the frame 322, while the top surface of each block Of course, the position can be slightly different, and by using software that calculates the displacement, the mass of fluid added to the well can be programmed to eliminate such differences. For the same reason, it is not essential that all blocks 328 have the same mass.

  Reagents and other fluids can be added to or removed from wells 317 in wells 348 that are removable from plate 300, thus replacing wells 348 during each use of plate 300. It goes without saying that the plate 300 may in principle be used multiple times and / or in multiple experiments, if possible.

  Reference is now made to FIGS. 4A to 4C showing a multiwell plate 400 and components thereof constructed and operative in accordance with embodiments taught herein. Specifically, FIG. 4A is a perspective view of the multi-well plate 400, FIG. 4B is an exploded view of the multi-well plate 400, and FIG. 4C is a multi-well view taken along section line IVC-IVC in FIG. 4A. 4 is a cross-sectional view of a plate 400. FIG.

  As shown in FIG. 4A, when assembled, the plate 400 appears generally similar to the plate 300 of FIGS. 3A-3G. However, it is also apparent from FIG. 4B and the description herein that the structure of plate 400 is somewhat different from plate 300, despite the similar purpose and application of plates 300 and 400.

  Since the plate 400 is designed for use with existing equipment, the dimensions are set according to the standard plate size currently used, and the wells are similarly spaced. Thus, when assembled, plate 400 comprises a plurality of wells 417 formed therein, with a plurality of wells 417 formed therein, with a plurality of side surfaces 416 between an upper surface 412, a lower surface 414, surfaces 412 and 414, and is similar to a typical 96 well plate. Looks.

  As shown in FIG. 4B, which is an exploded view of the plate 400, the plate 400 is actually formed of several parts. Side 416 is part of frame 422. The frame 422 also includes a well defining skeleton 450 that defines 96 bores 452 in the frame 422. Each bore has a substantially circular cross section along a horizontal plane and a substantially rectangular cross section along a vertical plane.

  The plate 400 further comprises 96 individual identical cylindrical well support elements 430, each with an opening 433, each dimensioned to fit with one of the bores 452, and a single well Suitable for receiving the defining element 419, the well defining element 419 can be easily removed from the well support element 430 receiving it. Each individual well 417 is defined by a well defining element 419 that is in the shape of a cylindrical portion 466 and is sealed at its bottom using a generally circular bottom piece 434.

  Each well support element 430 is glued or otherwise attached to two arms 426 and 426 located on plates 474 and 476, respectively. Arms 426 and 426 ′ are not required, but are integrally formed on plates 474 and 476. For clarity, subsequent arms 426 and 426 'are 426 unless specifically stated otherwise. The flexible arm 426 is a suitable material, typically a metal or plastic, with appropriate flexibility that can sense the addition or removal of small volumes to or from the well, as will be described in more detail later. It may be made.

  The arm 426 is substantially flat as shown in the inset of FIG. 4B. Each arm 426 and 426 ′ has a generally rectangular portion 426a with protrusions extending in its two adjacent corners, which are more than half of the circumference of the cylindrical well support element 430, but completely A partial ring 426b that does not extend is formed. The ring 426b is dimensioned to have an inner periphery that is slightly larger than the outer periphery of the cylindrical well support element 430. The ring 426c formed in the partial ring 426b is dimensioned to have the same inner periphery as the cylindrical well support element 430. Thus, the cylindrical well support element 430 is secured to the ring 426c, for example by gluing (the upper surface of the cylindrical well support element 430 is attached to the lower surface of the ring 426c of the arm 426 and the cylindrical well support element 430 is The lower surface of the support element 430 is attached to the upper surface of the ring 426c of the arm 426). This structure allows a pair of arms 426 and 426 'to hold the well support element 430 in place in the x and y axes, but with the arms 426 bent, the element and the z of the wells contained therein Allows movement along the axis.

  In order to maximize and localize the strain felt by the arms in the rectangular portion 426a and the partial ring 426b, the partial ring 426b and the rectangular portion 426a are made of a flexible material (e.g., metal or Plastic). In some embodiments, arms 426 and 426 ′ are thinner than plate bases 474 and 476, respectively, to facilitate bending. In some embodiments, the outer plate portions 474 and 476 of the arms 426 and 426 'may be attached to the frame 422 and / or the electronic card 446 (further details) to strengthen the plate portions 474 and 476 that do not require bending. Is attached to, or otherwise attached to (described below), the strain felt by the arm 426 is localized.

  In general, for fluids of several hundred microliters or less, it will be appreciated that only a few microliters or less of fluid is added to a given well, depending on the intended use. For example, less than 300 microliters (μl), less than 250 μl, less than 200 μl, less than 150 μl, less than 100 μl, less than 75 μl, less than 50 μl, less than 45 μl, less than 40 μl, less than 35 μl, less than 30 μl, less than 25 μl, less than 20 μl, less than 15 μl, 10 μl Less than, less than 5 μl, less than 4 μl, less than 3 μl, less than 2 μl, or less than 1 μl of fluid may be added to a given well. As a result, the flexible arm 426 must be properly flexible to flex in response to the addition (or removal) of such an amount (or equivalent mass) of fluid to (or from) the well.

  When assembled together, the outer periphery of the cylindrical well support element 430 is slightly smaller than the inner periphery of the bore 452, so a small gap 438 is provided between the well defining skeleton 450 and the cylindrical well support element 430. Is formed (see FIG. 4C). As will be described below, the presence of these gaps allows for independent movement of each well support element 430 along the vertical (z-) axis and concomitant deformation of the arms 426.

  To seal the gap 438 without hindering such vertical movement, a very thin (e.g., approximately 7 microns) flexible material (e.g., suitable plastic) film 440 is applied to the upper edge 442 of the frame 422. Attaching, in one embodiment, also attaching to the top card 446 (more details below). Film 440 includes 96 circular openings 443, which are openings 447 in card 446 (described below), ring 426d described in more detail below, and openings in the assembled well support element 430. It is aligned with 433.

  Film 440 is attached to frame 422 using any suitable means (eg, soldering, gluing, melting, bonding or any other suitable attachment mechanism). In the illustrated embodiment, it will be appreciated that each well defining element 419 can be easily removed from the well support element 430 in which it is located without damaging the structure and function of the sensitive arm 426. .

  Each arm 426 includes a pair of flat strain gauges 444 attached together at a rectangular portion 426a on the top surface thereof. Alternatively, the strain gauge may be attached to the upper and lower surfaces of the rectangular portion 426a. Each strain gauge is electrically connected to the electronic card 446 via, for example, a wire (not shown). In some embodiments, the arm 426 may be formed with a hole 445 to facilitate passing such wire therethrough. The electronic card 446 is arranged to minimize the connection length between the strain gauge 444 and the card.

  In one embodiment, the well support element 430 has a top and bottom portion, as shown in the enlarged portion of FIG. 4B rotated 90 degrees about the longitudinal axis of the well support element relative to the remainder of FIG. The bottom rim may be provided with a protective protrusion 431a, which is designed to protect the strain gauge 444. In addition, in some embodiments, the well support element 431 may include top and bottom protrusions 431b that are rotationally offset 180 degrees from the protrusions 431a. This protrusion 431b engages with the plates 474 and 476 and limits the range of bending of the arm 426.

  In one embodiment, the card 446 (which may actually be multiple cards) is electrically connected to a port 458 installed in the frame 422 via multiple wires (not shown), and FIG. As described below with respect to ˜9A, the ports are configured for connection to appropriately equipped plate bases and data reader electrical ports. In certain embodiments, the card 446 (which may actually be multiple cards) is connected to a USB port (not shown) installed in the frame 422 via multiple wires (not shown) or, for example, currently Electrically connected to similar input / output ports that are known or may be developed in the future, this port is configured for connection of the plate 400 to a power source and / or computing device. A data reader and / or computing device to which the plate 400 can be connected can implement the graphical user interface described below with respect to FIGS. Preferably, the port 458 and / or USB port (not shown) does not extend beyond the outer surface of the frame 422, and is preferably flush with the outer surface and affects the overall dimensions of the plate 400. Since it does not reach, standard equipment can be used.

  In some embodiments, the plate 400 further comprises a power supply (eg, a storage battery) (not shown) that connects to, for example, an electronic card 446, which power supplies the electronic components of the plate 400. If the plate 400 is connected to a power generation source or computing device via the port 458 or USB port, it can be recharged.

  Each electronic card 446 includes 96 openings 447 having a circular cross section, and when the plate 400 is assembled, the openings 447 align with the ring 426d, the section 452, and the well support element 430. An additional 96 smaller openings 448 are located near the opening 447. The opening 448 can pass through a wire (not shown) connecting the strain gauge 444 and the card 446.

  The electronic card 446 may have elements thereon for measuring the electrical resistance of the circuit (eg, Wheatstone bridge). The bending of the arm 426 causes a change in the length of the corresponding strain gauge 444 resistor. This change is correlated with a change in the electrical resistance of the circuit and is measured at an element on the electronic card 446 using, for example, a Wheatstone bridge. Changes in the electrical resistance of the circuit allow calculation of the mass of fluid added (or removed) to each well. Specifically, the greater the mass of fluid added to (or removed from) the well, the greater the change in flex of arm 426 and the greater the change in electrical resistance of the circuit. Therefore, the measured value of the change in the electrical resistance of the circuit indicates the change in the mass of the fluid in the well, and the change can be calculated based on the measured value. If the density of the fluid is known, calculation of the volume of fluid added (or removed) is facilitated. Thus, arm 426 with strain gauge 444 forms a signal provider that provides a signal that indicates a change in the amount of fluid in the well associated with it. In some embodiments, the electronic card 446 is a storage component for storing signals generated by a signal provider when, for example, the plate 400 is not connected to a data reader (e.g., the data reader of FIG. 8). Is provided. Such stored signals can then be retrieved from the card 446 by the data reader when connected to the plate 400.

  For example, if rows A to L of plate 400 are filled sequentially using an 8-chip dispenser, or if individual wells are filled sequentially, or if all wells are filled simultaneously, It is possible to calculate the amount of fluid added to the well, so that if the wrong amount of fluid is added to a particular row, either too much or too little, it will identify in real time column by column It becomes possible.

  Preferably, the device used to add fluid to the well comprises control software that the device can use to correct errors. In cases where too little fluid is added, additional fluid may be distributed to the affected wells so that the fluid in the well reaches the appropriate amount and / or the software may It may be possible to adjust the error by adding a reagent or reaction solution that is insufficient to the affected well. Similarly, if too much fluid is added to a particular well, it may be appropriately scaled up by adding reagents or reactants in further operations.

  Alternatively, the affected wells or well components may be included in further manipulations for the remainder of the experiment, and the results for a particular well will calculate the wrong volume used during the experiment. It may be used in the calculation after the end of the experiment by adjusting and eliminating. Yet another method is to identify abnormal wells with the wrong amount of fluid added to the wells, rather than discarding the results for all rows or columns or plates where the wells are located, after the end of the experiment. The abnormal well can be excluded from the calculation.

  Even if all wells are filled at the same time, the use of plate 400 in conjunction with the appropriate software to identify the addition of the wrong amount of fluid to a particular well support element 430 in real time will allow the user to It goes without saying that by allowing the ongoing experiment using the elements or wells of the plate 400 to be stopped, the waste of reagents and reactants in downstream experiments can be avoided.

  Furthermore, it will be appreciated that the plate 400 may operate without continuous monitoring. In such cases, baseline measurements of wells or plates are obtained from the signal provider. The plate may then be separated from the processor or data reader providing the signal and / or the power generation source and fluid added to or removed from the plate. The plate may then be reconnected to a power source and / or data reader or processor to obtain a second signal obtained from the signal provider. Comparison of the initial and second signals allows identification of specific wells where the wrong amount of fluid is present and can be excluded from further experiments and calculations.

  In some embodiments, the plate 400 is electrically coupled to the electronic card 446 near one or more wells 417 and is configured to provide an indication of temperature or temperature change. A temperature sensor (not shown) is also provided. Because temperature changes in the system can affect the strain gauge 444, knowledge of changes to temperature and computational considerations allow for more accurate identification of well weights and stabilization of well sample temperature It goes without saying that it will be possible to ensure reliability and increase sensitivity to temperature changes.

  In addition, the electronic card 446 has components for manipulating data collected by various sensor elements connected to the card (e.g., an analog-to-digital conversion configuration for converting Wheatstone bridge analog signals to digital signals). The elements and normalization components for normalizing the collected signal) may be electrically coupled.

  Since the bending of each arm can be calculated, the amount of substance added to each individual well or the volume of liquid can be calculated in real time, so the use of plate 400 can be used in further experimental operations to It makes it easy to modify the amount of material added to each well, or to ignore individual wells, rather than rows or whole plates. In addition, regarding this configuration, as will be described later with reference to FIG. 5A-7, it is possible to observe a decrease in the substance from a predetermined well over time.

  According to an embodiment of the invention, the plate 400 comprises means for heating and optionally cooling individual wells. Such heating means may be, for example, in the form of (a) a heating coil placed around at least some of the wells, or (b) a Peltier element (sometimes called a Peltier heat pump or thermoelectric cooler). Good. As is apparent, Peltier elements can be used to cool as well as heat individual wells, and can also detect or monitor the temperature of the wells or their surroundings. In this way, the temperature of individual wells can be controlled. For example, the temperature of each well can be maintained at 37 ° C. ± 0.5 ° C., respectively.

  In one embodiment, the Peltier element for each well may include one or both of the electronic cards 446 shown in FIG.4B (e.g., the heating of the well defining element 419 or a particular well support element 430 disposed in the opening). May be incorporated in the vicinity of each opening 447). Alternatively, heating coils or Peltier elements may be placed on some or each of the well support element 430 or well defining element 419 to heat the associated well or its interior. As yet another alternative, a heating coil or Peltier element may be used to heat a well associated therewith or its interior in the vicinity of a group of well defining elements 419 or well support elements 430 (e.g., to an electronic card 446). You may arrange. Referring to FIG. 4B, the provision of such well heating means is not limited to a plate in which the well can be displaced along the z-axis, but it goes without saying that such a heating means can be provided in a non-displaceable plate of the well. Nor.

  If the temperature of the well is measured periodically, and if the device used to measure the temperature is connected to a controller that controls the heating means for each individual well, then the temperature of the individual well is measured periodically It goes without saying that the individual well heating means used in conjunction with can provide a method for improving control over the conditions of a given well. Thus, for example, a plate 400 having such heating means is stored in a thermostat, the temperature of the well is periodically monitored, and the temperature of the individual well is heated (or cooled) as needed. You may adjust by doing. Alternatively, the heating means itself may be used for incubation, e.g., with frequent temperature monitoring and adjustment (i.e., every 15, 10 or 5 minutes, for example) to specify that incubation is to occur. The well temperature may be maintained at 37 ° C. ± 0.5 ° C., for example. Thus, by providing individual heating elements for each well in plate 400, well support element 430 or well defining element 419, the temperature can be adjusted and controlled if the temperature is found to be incorrect.

  Reagents and other fluids can be removed from the plate 400, added to or removed from the well defining element 419, which can optionally be discarded, replacing the well defining element 419 during each use of the plate 400 It will be appreciated that the plate 400 may be used multiple times and / or in multiple experiments if possible.

  Reference is now made to FIGS. 5A-5D, which are screen shots illustrating a graphical user interface for online (real-time) monitoring of fluid addition to a multi-well plate according to embodiments taught herein.

  As described above, the multi-well plate (e.g., multi-well plate 300 of FIGS. 3A-3G and multi-well plate 400 of FIGS. 4A-4C) according to an embodiment of the present invention is, for example, a suitable plate data reader (FIGS. May be electrically connected to the processor via a USB or other cable connected to a computing device (e.g., a computer). The graphical user interface of FIGS. 5A-5D operates on such a processor connected to the plate using data provided to the processor from the plate's electronic card (eg, electronic cards 546 and 646). The processor may be configured to provide only graphical user information or may be configured to control the amount of fluid dispensed to the well.

  It will be appreciated that the data for providing on-line monitoring is dependent on baseline electrical resistance measurements measured by plate strain gauges (eg, strain gauges 344 and 444). This baseline measurement generally corresponds to an empty well when monitoring well filling. Once fluid is dispensed into the wells, the electronic card facilitates the graphical user interface functions described below by providing the processor with an indication of the volume of fluid added to each well. In some embodiments, the analog data representing the baseline electrical resistance is normalized and converted to digital data by an appropriate element located on the plate's electronic card. As a result, the processor accepts data suitable for use with a graphical user interface.

  As shown, the graphical user interface 500 is associated with experiment design software (not shown) and includes specific details of the experiment to be performed (eg, experiment name, substance used, volume of liquid dispensed, upper limit of capacity). The lower limit, required accuracy (ie, experimental sensitivity) and any other suitable experimental parameters) are shown in the information box 502 of the graphical user interface. As described below, the details contained in the information box 502 provide the user with a baseline indication that is used to alert the user of the erroneous addition of a substance to any one or more wells. Of course, in some embodiments, the software that operates the fluid dispensing device can calibrate the amount dispensed and the degree of sensitivity used based on these experimental parameters.

  The graphical user interface 500 further comprises a graphical representation 504 of a multiwell plate in which experiments are currently being performed based on information provided from the plate's electronic card. The plate graphic representation 504 comprises a plurality of circles 506, each of which is a plate well (shown here as a 96-well plate), and a plate row or column index, identified by reference numerals 508 and 510, respectively. Corresponding to

  The purpose of the additional online monitoring of fluid to the plate is to facilitate real-time control of the volume of fluid added to the plate. Graphic user interface 500 allows the user to monitor real-time conditions and if the process control is not fully automated, after initial entry of parameters, the fluid volume may be modified as required or predetermined The system can be instructed to take other steps to compensate for the wrong volume of the well. In general, the graphic user interface further provides a graphic indication of whether a target fluid volume has been reached or whether additional fluid should be added to the plate. In some embodiments (e.g., those shown in FIGS. 5A-5D), the graphic indicator has a fill pattern or color indicator, the first fill pattern or color indicates that the well is empty, and the second fill The pattern or color represents a well whose fluid volume is less than the target fluid volume, and the third fill pattern or color is the fluid volume appropriate within a certain tolerance and equal to the target fluid volume Represents a well. In some embodiments, a fourth fill pattern or color is used to represent wells that exceed the target fluid volume.

  FIG. 5A illustrates a graphical user interface 500 prior to the start of the experiment. Thus, not all circles 506 are filled patterns (or first colors). This indicates that the well is empty.

  FIG. 5B illustrates the graphical user interface 500 as fluid distribution into the plate row 1 begins. Thus, in the graphic display 504, the circles 512 corresponding to the wells in column 1 (A1, B1, C1, D1, E1, F1, G1, and H1 wells) are diagonal lines inclined from right to left here. The second fill pattern (or second color) shown indicates that the fluid volume in the well is less than the target volume. The remaining circles 506 corresponding to the wells in columns 2-12 remain in a pattern or color that clearly indicates that the well is empty.

  In some embodiments (not shown), the graphical display 504 demonstrates the volume of fluid that must be added to the well in order to reach the target fluid volume in the well. For example, this may be accomplished by scrolling the pointer over one of the circles 506 as controlled by a computer mouse, thereby indicating the volume that the pop-up box should add to the corresponding well.

  In FIG. 5C, as fluid distribution to the wells in row 1 continues, most of the circles 512 correspond to the third fill pattern (or third color) (here the third color) corresponding to the well that has reached the target fluid volume. It can be seen that it is represented as a dense diagonal line sloping from left to right. Graphic display 504 indicates that well C1 has not yet reached the target fluid volume, and circle 514 corresponding to well C1 remains a diagonal second fill pattern (or the same color) that slopes from right to left. It is an additional indication by that.

  Alternatively, the amount of fluid in the well can only be dispensed after the fluid has been dispensed into the well (e.g., if the liquid is dispensed continuously but slowly or in droplets, continuous or multiple as the well is filled). In an embodiment that is determined (as opposed to taking measurements), the graphical display 504 does not provide information as shown in FIG. 5B. Rather, once the fluid has been dispensed, the graphical display 504 shows which well has a smaller (or more) volume of fluid than is required in a manner similar to that shown in FIG. Show.

  FIG. 5D is the same as FIG. 5C, but the graphical user interface after distribution of additional fluid volume to well C1 to correct the initial shortage so that the volume of fluid in well C1 is equal to the target volume of the experiment Is shown. When all wells in column 1 are correctly filled with the target fluid volume, the corresponding circle 512 is a dense diagonal third fill pattern that slopes from left to right indicating that the target volume has been reached (or 3rd color).

  As fluid is dispensed into the wells in the additional rows of the plate, the graphical display 504 changes so that the color of each circle 506 indicates the volume of fluid in the corresponding well. This provides a real-time indicator of the volume of fluid in each well and prevents errors in the volume of fluid added to the well.

  In some embodiments, fluid is dispensed simultaneously into all wells 506 of the plate, one at a time, whether continuously slow or continuously rapid. In such an embodiment, the graphical display 504 provides an indication similar to that shown in FIGS. 5B, 5C, and 5D for all wells simultaneously, rather than column by column, as described above.

  Reference is now made to FIGS. 6A and 6B, which are screenshots showing a graphical user interface for offline volume monitoring of fluids in a multi-well plate according to embodiments taught herein.

  The graphical user interface 600 of FIGS. 6A and 6B is a plate data reader or USB, or other electronic connector (e.g., electronic card 346 and electronic card) currently known or may be developed in the future. 446) is similar to the graphical user interface 500 of FIGS. 5A-5D in that it operates on a processor connected to the plate using data provided to the processor. Similarly, the graphical user interface 600 includes a graphical representation 604 of a multi-well plate currently performing an experiment based on information provided from the plate's electronic card. The plate graphic representation 604 comprises a plurality of circles 606 (each corresponding to a plate well (here, a 96-well plate)) and a row or column indicator of the plate clearly indicating reference numerals 608 and 610.

  However, during off-line monitoring, rather than showing the user whether the proper fluid volume has been dispensed to the well, for some reason the volume of fluid in the well is lower than the target volume or predetermined The goal is to provide an alarm if the threshold is below. Thus, the graphical user interface 600 is not associated with the experimental design software, but rather is an indication that the user should receive an inappropriate volume of wells (e.g., an alarm or action to take corrective action). Enter a value for audible or document notification. In some embodiments, the value is a predetermined absolute value for capacity. Then, when the volume of fluid in the well is lower than a predetermined volume, the graphical user interface 600 provides an indication of a particular well with a low volume. In certain embodiments (eg, the illustrated embodiment), the value is a change value for capacity. The graphical user interface 600 then provides an indication when the volume of fluid in the well changes beyond a predetermined value. In this variation (not shown), an indicator may be provided if the value exceeds a certain percentage and falls below a predetermined baseline. The value used to provide an indication to the user may be an initial value and may be set by the user based on experimental sensitivity or other appropriate considerations.

  As shown in FIGS. 6A and 6B, different magnitude volume changes are manifested by different fill patterns or colors, and legend 612 corresponds to how much fluid corresponds based on the fill pattern or color of circle 606. Provided so that the user can identify what is missing from the well. In the illustrated embodiment, the diagonal fill pattern indicates that the volume of fluid in the corresponding well is unchanged and equal to the initial volume, and the dot fill pattern indicates that the volume of fluid in the corresponding well is less than 1 microliter. The check pattern fill pattern shows the change when the volume of the fluid in the corresponding well is 1 microliter or more and less than 3 microliters.

  The graphical user interface 600 further comprises one or more graphs 614, where the volume of fluid in one or more particular wells can be plotted as a function of time. In some embodiments, the particular well whose information is shown in graph 614 is displayed by the user (e.g., by pointing the particular well with a mouse cursor or by identifying the well with an appropriate text box (not shown)). ) May be selected). In some embodiments, information corresponding to each of the wells may be displayed sequentially and / or repeatedly in the graph 614.

  In general, the data reader or USB or other connector associated with the electronic card on the plate is used at a certain frequency (e.g. once daily, once every hour, once every minute, once every 30 seconds or even more Reports the measured volume of each well to the processor at once per second). The exact frequency may be coded on an electronic card at the factory or set by the user according to the requirements of the experiment to be performed.

  Referring to FIG. 6A, at the first time point T1, no change in fluid volume is seen in any well, so all circles 606 are filled with a fill pattern (diagonal line) corresponding to the nominal volume. I can see that. None of the wells are plotted in graph 614 because there is no change in fluid volume.

  In FIG. 6B, which shows the graphical user interface at the second time point T2 after T1, the fill pattern in which the circles 616 corresponding to wells C1, D1, and E1 indicate the change in fluid volume in the wells below 1 microliter Circles 618 corresponding to wells A4, B1, F1, and G1 are filled with (dots), and indicate a change when the fluid volume in the well (check pattern) is 1 microliter or more and less than 3 microliters You can see that it is filled with a pattern. In the illustrated example, graph 614 depicts a plot 620 of the change in volume of well A4 as a function of time based on multiple readings of the volume of fluid in well A4.

  Reference is now made to FIG. 7, which is a screenshot showing a graphical user interface for offline temperature monitoring of fluids in a multi-well plate in accordance with embodiments taught herein.

  The graphical user interface 700 of FIG. 7 is similar to the graphical user interface 600 of FIGS. 6A and 6B, except that the fill pattern (or color) of the circle 706 forms part of the plate. The point of representing the temperature of the corresponding well in the plate, as measured by the temperature sensor, and the point or points 714 can include a plot of the temperature in one or more specific wells as a function of time. Different from the graphical user interface 700.

  In some embodiments (e.g., those shown in FIG. 7), there are temperature sensors associated with each well, while in other embodiments there are fewer temperature sensors than wells, but still a large number of temperature sensors. May be present.

  As shown in FIG. 7, different temperatures are indicated by different fill patterns (or colors), and the number of wells or their vicinity is determined by the user based on the fill pattern (or color) of the circle 706. A legend 712 is provided so that can be identified.

  In FIG. 7, at time T1, it can be seen that each well is at a specific temperature as indicated by the fill pattern of the corresponding circle 706. For example, the filling pattern of the circle 706 corresponding to the well E1 indicates that the temperature of the well E1 is 39 ° C.

  In the example shown, graph 714 has a plot 720 showing the change in temperature in well A5 as a function of time.

  In general, a data reader or USB or other connector associated with a plate temperature sensor is used at a certain frequency (e.g. once daily, once every hour, once every minute, once every 30 seconds or even Report the temperature at or near each well to the sensor once a second). The exact frequency may be coded on an electronic card at the factory or set by the user according to the requirements of the experiment to be performed. Thus, the fill pattern (or color) of the circle 706 in the graphical user interface 700 changes when the data reader indicates a change in the temperature of the corresponding well as measured by the temperature sensor.

  Since the well-providing element 348 and the well-defining element 419 are removable from their respective plates 300 and 400, as described above, the element 348 or the element 419 can be used to obtain measurements, the element 348 or Removing elements 419 and inserting them into another simpler plate (not shown) lacking the detector detailed above for storage (eg for storage in a refrigerator or incubator) it can. Later, if another measurement is desired, element 348 or element 419 may be reinserted into plate 300 or 400, respectively.

  Reference is now made to FIG. 8, which is a perspective view of a data reader 800 and plate base constructed and operative in accordance with one embodiment taught herein to receive signals from a multi-well plate in accordance with an embodiment of the present invention.

  As shown in FIG. 8, the base plate and data reader 800 includes a base 802, and the base 802 forms a frame 810 thereon, and a multiwell plate (e.g., the plate 200, 300 or 400 described above) is formed therein. Optimal shape and size to accommodate. In one embodiment, the base plate and data reader 800 is an optical machine or imaging device (e.g., a Hermes system commercially available from Idea Bio-Medical Ltd., Rehovot, Israel) (http: //www.idea-bio. com / page-87 -__ Hermes.aspx)). In some such embodiments, the base 802 transmits at least some wavelength of illumination so that the plate is positioned with the data reader 800 and the sample of the multi-well plate can be imaged by an optical machine or imaging device. May be.

  In some embodiments, the frame 810 includes a holding mechanism that holds the plate stably and securely within the data reader 800. In some embodiments, the retention mechanism comprises a protrusion 812 that engages the frame of the plate, and this protrusion 812 may be retractable in the frame 810 (eg, with a spring force). As described above, when the user inserts the plate into the data reader 800, the user presses the plate against the protrusion 812 so that the protrusion 812 is accommodated in the frame 810. Once the user stops pressing the plate, for example, when the plate is in place, the spring protrudes outward so that the protrusion 812 engages the plate and holds it in the data reader 800. Press object 812. In some embodiments, the retention mechanism comprises a mechanism for snapping the plate to the data reader 800, a rim that can stop the plate, and the like.

  In some embodiments, the frame 810 includes a recess 814 to assist the user in gripping the plate disposed within the frame 810 if the user wishes to remove the plate. Other mechanisms that facilitate removing the plate from the frame 810 (eg, an eject button) may be used.

  Frame 810 is an electrical port 820 arranged and configured to electrically engage a corresponding port on a plate located in the data reader (e.g., port 358 in FIG. 3B or port 458 in FIG.4B). Is further provided. The electrical port 820 is for use of a processor (not shown) for providing information from the plate to the processor (eg, using dedicated software (eg, experimental design software or graphical user interface software described above with reference to FIGS. 5A-7)). May be electrically connected).

  Here, a perspective view of a device for removing a well-providing element or well-defining element and / or placing such element in a multi-well plate from a device constructed and operative in accordance with one embodiment taught herein. See 9A and 9B.

  As shown in FIGS. 9A and 9B, a device 900 for operation of a well-providing element (e.g., element 330) or a well-defining element (e.g., element 419) is functionally associated with a plate holding base 902 and plate holding The base 902 is configured to place the multiwell plate 904 therein. As shown in FIGS. 9A and 9B, the plate holding base 902 may be a data reader and base (for example, the data reader and table 800), or may be a simple base on which a multiwell plate is placed.

  A well engaging portion 906 movably mounted on the vertical displacement mechanism 908 is configured on the plate holding base 902. The vertical displacement mechanism 908 is configured to allow vertical displacement of the well engaging portion 906 toward and away from the plate 904 disposed on the plate holding base 902. In some embodiments, the vertical displacement mechanism 908 includes a vertical mount 910 and a displaceable portion 912 that is vertically displaceable along the mount 910. Accordingly, the well engaging portion 906 is mounted on the displaceable portion 912 and can be displaced together therewith.

  A plurality of well-engaging protrusions 916 are disposed on the lower surface 914 of the well-engaging portion 906, each being a well-providing element or a well-defining element that is installed on or removed from the plate 904 for its attachment. Configured to fit one of the wells. In some embodiments, the well engagement protrusion 916 engages the corresponding well by a snap fit mechanism, but other engagement methods (eg, reduced pressure) are also contemplated.

  With respect to the placement of the well defining element or well-providing element on the plate 904, the well engaging protrusion 916 engages the well defining element or well-providing element, after which the well engaging portion 906 is the well defining element or well-providing element. Are displaced vertically toward the plate 904 until they fit in their proper position within the plate 904 (eg, within the section 352 of the plate 300 or within the well support element 430 of the plate 400). Next, the well engagement protrusion 916 moves away from the well defining element or well-providing element, and the well engagement portion 906 moves vertically away from the plate 904 so that the element is properly installed in the plate 904 Access is possible for insertion of the reagent into it.

  With respect to the removal of the well defining element or well-providing element from the plate 904, the well engaging portion 906 is moved until the well defining element or well-providing element installed in the plate 904 engages the well coupling protrusion 916. Displace vertically to. The well engagement protrusion 916, along with the well engagement portion 906 and the well engaged therewith, is vertically displaced away from the plate 904 to remove the well defining element or well-providing element from their position within the plate 904. It is. When the well engaging portion is sufficiently displaced from the plate 904, the well engaging protrusion 916 is then moved away from the well defining element or well-providing element.

  It will be appreciated that similar devices may be used to engage pipette tips and the like, and may be used to dispense fluid (eg, reagents) into the wells of plate 904.

  Variations of structures that may be employed with multi-well plates (e.g., those described herein with reference to FIGS. 2A-4C) and other multi-well plates in certain cases, and methods of using those plates The variations are described below.

  The plate described above utilizes the physical displacement of the well along the z-axis to determine the volume of fluid dispensed into one or more wells. In the embodiments described above, such physical displacement is distributed to the wells during observation (or wells (if density and mass occupy a determinable volume)) (or wells). Connected to a strain gauge to induce a signal that correlates with the volume of fluid (lost from). However, it will be appreciated that other methods may be used instead of or in addition to the use of a strain gauge to determine the capacity.

  Thus, for example, if a multi-well plate with wells that are displaceable in the z-axis is used in conjunction with a reading device having an autofocus mechanism, it may be used to determine the amount of well displacement. An example of such an autofocus mechanism is described in US Pat. No. 7,109,459 whose title is “Auto-focusing method and device for use with optical microscopy”. The contents shall be quoted.

  For illustration, a multi-well plate with displaceable wells is introduced into a reading device with an autofocus mechanism (e.g., a Wiscan® scanner available from Idea Bio-Medical Ltd., Rehovot, Israel). May be. By combining the autofocus mechanism and appropriate feedback control, the bottom of the well of interest can be set to the same height before dispensing the fluid. The fluid may then be dispensed into the wells. In some cases, it may be done online without moving the plate to another position, and in other cases it may be necessary to move the plate to the dispensing station. After dispensing the fluid, the autofocus mechanism may be used again at this time to determine the z-axis well behavior (returning the plate to the autofocus position takes precedence if necessary). As described above, this information can be used in turn to determine the amount of fluid dispensed into one or more wells. In addition, measurements may be taken periodically to determine whether fluid has been lost (eg, due to evaporation) from one or more wells. As described above, this method may be used with individual wells or groups of wells. As described above, this method may be used in conjunction with or instead of a strain gauge to determine the displacement of one or more wells.

  As in the method already described above, the detection of the wrong amount of fluid in one well or group of wells may be the correction of the amount of fluid in the well, the exclusion of the well from further manipulation and / or calculation, or In some cases, the calculation is easily corrected.

  Another method for determining the amount of fluid lost from a well involves periodic monitoring of the temperature of the individual wells. Often, multi-well plates containing live cells are incubated at 37 ° C. However, the heat distribution of the incubator can be uneven, and other factors can cause a non-uniform temperature distribution in the plate, which causes various reductions in fluid from various wells, It can adversely affect the cells in it. Determining the amount of fluid lost from a well over time by tracking the temperature of individual wells periodically (e.g. once per hour) and considering the nature of the fluid in the well Not only is it possible, but it is also possible to modify the temperature of the well as described above, for example using heating or cooling means. Such monitoring may be facilitated by placing individual temperature sensors in each well (eg, at the bottom or sides thereof). Such sensors may be electronically connected to a card (eg, 346 or 446 described above) to facilitate data reader reading. Alternatively, the temperature of individual wells may be detected periodically using one or more temperature cameras.

  Similar to the method already described above, the detection of an incorrect amount of fluid in a well or group of wells may be the correction of the amount of fluid in the well, the exclusion of the well from further manipulation and / or calculation or In some cases, the calculation is easily corrected. If the well of the plate is displaceable along the z-axis, this method can be used in conjunction with a strain gauge, as described above, or alternatively, and in conjunction with a method using an autofocus mechanism, Alternatively, it may be used instead. However, it goes without saying that, unlike methods using strain gauges or autofocus, this method may be used with plates in which the wells cannot be displaced.

  It goes without saying that the embodiments shown in the figures are for illustration only and that these variations are within the scope of the invention. For example, the number of wells per plate, the shape of the wells, and the materials used may differ from those shown or specifically described herein, such as detecting, adding or The removal means is the same. In addition, each of the wells may be provided with an additional layer or insert (eg, a well insert that grows cells). The osmotic reagent can then be added to the well environment without directly participating in the cells growing in the insert.

  For clarity, it will be appreciated that certain features of the invention described in the context of separate embodiments may be provided in combination in a single embodiment. On the contrary, for the sake of brevity, the various features of the invention described in the context of a single embodiment may be separated separately, in any appropriate subcombination, or as appropriate, in any other It may be provided in the embodiments of the invention described. Certain features that are described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperable without those elements.

  Although the invention has been described with reference to specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, all such alternatives, modifications and variations are intended to be included within the scope of the appended claims.

  Citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention.

  Paragraph headings are used herein to facilitate understanding of the specification, but should not be construed as essential restrictions.

Claims (67)

A plate comprising a first substantially planar surface, a second substantially planar surface, at least one well and at least one signal provider;
The first substantially planar surface comprises at least one first opening defined therein;
The second substantially planar surface is substantially parallel to the first substantially planar surface;
The second substantially planar surface is spaced from the first substantially planar surface;
The at least one well is defined in the plate;
The at least one well includes a second opening corresponding to and aligned with one of the at least one first opening;
The at least one well comprises a sidewall and a bottom;
The at least one well extends from the first substantially planar surface toward the second substantially planar surface;
The at least one well is displaceable away from the first substantially planar surface;
The at least one signal provider is functionally associated with the at least one well;
A plate capable of providing a signal in response to displacement of the at least one well away from the first surface. The first surface comprises a plurality of first openings defined therein;
A plurality of wells are defined in the plate;
Each well includes a second opening corresponding to and aligned with the first opening of the plurality of first openings defined by the first surface;
Each well has a sidewall and a bottom,
The plate of claim 1, wherein each well extends from the first substantially planar surface toward the second substantially planar surface.   The plate of claim 2, wherein each of the plurality of wells is displaceable away from the first substantially planar surface. A multi-well plate comprising a first substantially planar surface, a second substantially planar surface, a plurality of wells and at least one signal provider,
The first substantially planar surface comprises a plurality of first openings defined therein;
The second substantially planar surface is substantially parallel to the first substantially planar surface;
The second substantially planar surface is spaced from the first substantially planar surface;
The plurality of wells are defined in the plate;
Each well has a second opening corresponding to and aligned with one of the first openings defined by the first surface;
Each of the wells comprises a sidewall and a bottom;
Each well extends from the first substantially planar surface toward the second substantially planar surface;
Each of the wells is displaceable away from the first substantially planar surface;
The at least one signal provider is functionally associated with the plurality of wells;
A multi-well plate capable of providing a signal in response to displacement of at least one of the wells away from the first surface.   5. A plate according to any preceding claim, wherein the first and second surfaces are spaced apart by a plurality of sidewalls extending between the first and second surfaces.   All of the well movements are connected so that the signal provider can provide a single signal in response to any one or more displacements of the wells. Crab plate. Some of the well movements connect to two or more groups,
The at least one signal provider comprises a multi-signal provider;
6. A plate according to any of claims 2-5, each capable of providing a signal in response to one displacement of the group. The at least one signal provider comprises a plurality of signal providers;
Each is associated with one well of the plurality of wells;
Each of the plurality of wells is displaceable independently of the other to move away from the first surface;
6. A plate according to any of claims 2 to 5, wherein each of the plurality of signal providers is capable of providing a signal in response to a displacement of one of the plurality of wells associated therewith.   The at least one signal provider includes 300 milligrams of material, 250 milligrams of material, 200 milligrams of material, 150 milligrams of material, 100 milligrams of material, 75 milligrams of material, 50 milligrams of material, 45 milligrams of material in the well. 40 mg substance, 35 mg substance, 30 mg substance, 25 mg substance, 20 mg substance, 15 mg substance, 10 mg substance, 5 mg substance, 4 mg substance, 3 mg substance 9. A signal can be provided in response to the placement of 2 milligrams of material or 1 milligram of material, 500 micrograms (μg) of material, 300 μg of material, 200 μg of material or 100 μg of material. A plate according to any one of the above.   The at least one signal provider includes 300 microliters (μl), 250 μl fluid, 200 μl fluid, 150 μl fluid, 100 μl fluid, 75 μl fluid, 50 μl fluid, 45 μl fluid, 40 μl fluid in the well, 35 μl fluid, 30 μl fluid, 25 μl fluid, 20 μl fluid, 15 μl fluid, 10 μl fluid, 5 μl fluid, 4 μl fluid, 3 μl fluid, 2 μl fluid, 1 μl fluid, 0.5 μl fluid, 0.3 10. A plate according to any of claims 1 to 9, capable of providing a signal in response to an arrangement of [mu] l fluid, 0.5 [mu] l fluid, or 0.1 [mu] l fluid.   11. A plate according to any of claims 1 to 10, wherein the signal provider is capable of providing a signal in response to displacement of at least one of the wells towards the first surface.   The at least one signal provider includes 300 milligrams of material, 250 milligrams of material, 200 milligrams of material, 150 milligrams of material, 100 milligrams of material, 75 milligrams of material, 50 milligrams of material, 45 milligrams of material in the well. 40 mg substance, 35 mg substance, 30 mg substance, 25 mg substance, 20 mg substance, 15 mg substance, 10 mg substance, 5 mg substance, 4 mg substance, 3 mg substance 12. A signal can be provided in response to removal of 2 milligrams of material or 1 milligram of material, 500 micrograms (μg) of material, 300 μg of material, 200 μg of material or 100 μg of material. A plate according to any one of the above.   The at least one signal provider includes 300 microliter (μl) fluid, 250 μl fluid, 200 μl fluid, 150 μl fluid, 100 μl fluid, 75 μl fluid, 50 μl fluid, 45 μl fluid, 40 μl of the well Fluid, 35 μl fluid, 30 μl fluid, 25 μl fluid, 20 μl fluid, 15 μl fluid, 10 μl fluid, 5 μl fluid, 4 μl fluid, 3 μl fluid, 2 μl fluid, 1 μl fluid, 0.5 μl fluid 13. A plate according to any of claims 1 to 12, which is capable of providing a signal in response to removal of 0.3, 0.3 [mu] l fluid, 0.2 [mu] l fluid, or 0.1 [mu] l fluid.   14. A plate according to any preceding claim, wherein at least one well in the plate is removable therefrom.   15. A plate according to any preceding claim, wherein the plate comprises at least one temperature sensor associated with at least one of the wells.   16. A plate according to claim 15, wherein the temperature sensor is placed on or in one of the wells.   17. A plate according to any of claims 15 or 16, wherein the at least one temperature sensor is configured to provide a signal representative of the temperature of the at least one well or its proximity.   18. The at least one temperature sensor is configured to continuously detect the temperature of the at least one well and periodically provide the signal representative of the temperature. Plate.   19. An electronic storage element for storing at least one signal provided by at least one of the at least one signal provider and the at least one temperature sensor. Plate.   20. A plate according to any of claims 15 to 19, wherein the at least one temperature sensor is configured to detect the temperature in a group of wells from the plurality of wells.   20. A plate according to any of claims 15 to 19, wherein the at least one temperature sensor comprises a single temperature sensor configured to detect the temperature of all the wells. The at least one temperature sensor comprises a plurality of temperature sensors;
20. Each of the claims 15-19, each associated with the one of the plurality of wells to detect the temperature in one of the plurality of wells associated therewith. Plate as described. Further comprising at least one heating component associated with the at least one well;
23. A plate according to any preceding claim, wherein the at least one heating component is located sufficiently close to the at least one well to heat the at least one well or the interior thereof. The at least one heating component comprises a plurality of heating components;
Each associated with the one well of the plurality of wells to heat one well or the interior thereof without substantially heating other portions of the plurality of wells, and 24. A plate according to claim 23, placed sufficiently close to the associated well.   25. A plate according to claim 23 or 24, wherein the at least one heating component comprises a heating coil.   25. A plate according to claim 23 or 24, wherein the at least one heating component can also cool the at least one well.   27. A plate according to claim 26, wherein the heating component comprises a Peltier element.   28. The at least one well in the plate is removable therefrom without removing the heating component associated with the at least one removable well from the plate. plate. At least one well in the plate is removable therefrom;
28. A plate according to any of claims 23 to 27, wherein the heating component associated with the at least one removable well is attached or integrally formed with the at least one well and is removable therewith. .   30. A plate according to any preceding claim, further comprising an electrical port operatively associated with at least one of the at least one signal provider and the at least one temperature sensor. It further includes a rechargeable power supply,
The rechargeable power supply is functionally associated with at least one of the at least one signal provider and the at least one temperature sensor and is recharged by connecting it to a power generation source. The plate according to any one of claims 1 to 30, wherein the plate is configured as follows.   32. The plate of claim 31, wherein the rechargeable power supply is configured to be recharged when the electrical port is electrically connected to the power generation source. A data reader,
The data reader contains a plate according to any of claims 30 to 32 therein,
The data reader comprises a base, an electrical port, and a processor,
The base is for installing a plate there;
The electrical port corresponds to the electrical port of the plate for electrical engagement therewith;
A data reader, wherein the processor is functionally associated with the electrical port to process a signal obtained from at least one of the signal provider and the temperature sensor via the electrical port.   34. The data reader of claim 33, wherein the processor is configured to obtain a signal directly from at least one of the signal provider and the temperature sensor via the electrical port.   34. The processor of claim 33, wherein the processor is configured to obtain the signal from an electronic storage component that stores at least one signal provided by at least one of the signal provider and the temperature sensor. Data reader. Further comprising a display functionally associated with the processor;
36. A data reader according to any of claims 33 to 35, wherein the display is configured to provide a user with information obtained from the processed signal. The information is
The amount of fluid in the plate at a particular time;
The amount of fluid in at least one of the wells at a particular time; and
Change in the amount of fluid in the plate over time;
A change in the amount of fluid in at least one of the wells over time;
The temperature of at least one said well at a particular time; and
37. The data reader of claim 36, comprising at least one indication of a change in temperature of at least one of the wells over time. The processor is configured to process the signal in real time;
38. The data reader of claim 37, wherein the display is configured to provide the information in real time to the user.   The data reader according to any one of claims 22 to 38, wherein the plate according to any one of claims 1 to 21 and 40 to 48 is arranged in the data reader. A plate comprising a first substantially planar surface, a second substantially planar surface, at least one well and at least one heating component;
The first substantially planar surface comprises at least one first opening defined therein;
The second substantially planar surface is substantially parallel to the first substantially planar surface;
The second substantially planar surface is spaced from the first substantially planar surface;
The at least one well is defined in the plate;
The at least one well includes a second opening corresponding to and aligned with one of the at least one first opening defined in the first surface;
The at least one well comprises a sidewall and a bottom;
The at least one well extends from the first substantially planar surface toward the second substantially planar surface;
The at least one heating component is associated with the at least one well;
A plate placed sufficiently close to the at least one well to heat the at least one well or the interior thereof. The first surface comprises a plurality of first openings defined therein;
The at least one well comprises a plurality of wells defined in the plate;
Each well includes a second opening corresponding to and aligned with one first opening from the plurality of first openings defined in the first surface;
Each of the wells comprises a sidewall and a bottom;
Each well extends from the first substantially planar surface toward the second substantially planar surface;
Since the at least one heating component comprises a plurality of heating components, each of the wells comprises one of the plurality of heating components associated therewith,
41. Each of the heating components is placed sufficiently close to the well with which the heating component is associated to heat the well or its interior without substantially heating other wells. Plate as described in.   42. A plate according to claim 40 or 41, wherein the heating component comprises a heating coil.   42. A plate according to claim 40 or 41, wherein the heating component can also cool the well.   44. A plate according to claim 43, wherein the heating component comprises a Peltier element.   45. The at least one well in the plate is removable therefrom without removing the heating component associated with the at least one removable well from the plate. plate. At least one well of the plate is removable therefrom;
45. The heating component associated with the at least one removable well is attached to or integrally formed with the at least one well and is removable therewith. Plate.   47. A plate according to any of claims 40 to 46, wherein the plate is also a plate according to any of claims 1-32.   47. A plate according to any of claims 40 to 46, wherein the plate is not a plate according to any of claims 1-32. A method for measuring the amount of fluid applied to a plate according to any of claims 1-32 and 47, comprising:
The method
Recording an initial signal provided by the signal provider;
After adding fluid to at least one well in the plate, obtaining a second signal generated by the signal provider in response to the addition of the fluid; and
A method wherein the amount of fluid added to the at least one well can be calculated based on the difference between the initial signal and the second signal.   50. The method of claim 49, further comprising adding fluid to the plate after the step of recording an initial signal and before the step of obtaining a second signal.   51. The method of claim 49 or 50, further comprising calculating an amount of the fluid added to the at least one well based on the difference between the initial signal and the second signal.   The step of calculating an amount comprises: (a) a volume of the fluid added to the at least one well; (b) a volume of the fluid added to the at least one well; and (c) the at least one well. 52. The method of claim 51, comprising calculating at least one of the volume and mass of the fluid added to a well. A method for measuring the amount of fluid lost from a well-equipped plate according to any of claims 1-22 and 47,
The plate has an initial amount of fluid dispensed into at least one well of the plate;
Recording an initial signal provided by the signal provider at a first time;
Obtaining a second signal from the signal provider at a second time after the first time;
Calculating the amount of fluid lost from the at least one well of the plate based on the difference between the initial signal and the second signal.   The step of calculating an amount comprises: (a) a volume of the fluid lost from the at least one well; (b) a mass of the fluid lost from the at least one well; (c) the at least one well. 54. The method of claim 53, comprising calculating at least one of the volume and mass of the fluid lost from the. Periodically repeating said step of obtaining a signal;
Calculating the amount of fluid lost from the at least one well in a period between the two different times based on a difference between signals obtained at two different times. 55. The method according to 53 or 54. Obtaining a baseline measurement of the displacement of at least one well in the multi-well plate;
Obtaining a second measurement of the displacement of the at least one well at a time after the step of obtaining the baseline measurement;
Calculating a change in the amount of fluid in the at least one well based on the second measurement of displacement, and
The multi-well plate has disposed therein a displacement measurement assembly for measuring displacement of at least one well in the plate in response to a change in the amount of fluid in the at least one well;
The baseline measurement is obtained via the displacement measurement assembly Obtaining a baseline measurement of displacement of at least one well in the multi-well plate at a first time; and
Measuring the displacement of the at least one well at a second time after the first time;
Calculating a change in the amount of fluid in the at least one well based on a change in displacement between the first and second times; and
The multi-well plate has disposed therein a displacement measurement assembly for measuring displacement of at least one well in the plate in response to a change in the amount of fluid in the at least one well;
The baseline measurement is obtained via the displacement measurement assembly;
The method wherein at least one of the first and second times has a detectable amount of fluid present in the well. Periodically repeating the step of measuring the displacement of the at least one well at a second time; and
Calculating a change in the amount of fluid in the at least one well during a period between the two measurements of displacement based on a change in displacement between the two measurements of displacement; and 58. The method of claim 57, comprising:   59. A method according to any of claims 56 to 58, wherein the change in the amount of fluid is due to the addition of fluid to the at least one well.   59. A method according to any of claims 56 to 58, wherein the change in the amount of fluid is due to a decrease in fluid from the at least one well.   61. A method according to any of claims 56 to 60, wherein the plate is a plate according to any of claims 1-32 and 47.   The signal provider is the volume of fluid in the at least one well and is 300 microliters (μl), 250 μl, 200 μl, 150 μl, 100 μl, 75 μl, 50 μl, 45 μl, 40 μl, 35 μl, 30 μl, 25 μl, 20 μl, 15 μl, 10 μl Any of claims 51 to 61, sensitive enough to detect changes in 5 μl, 4 μl, 3 μl, 2 μl, 1 μl, 0.5 μl fluid, 0.3 μl fluid, 0.2 μl fluid or 0.1 μl fluid The method of crab.   The signal provider is 300 milligrams (mg), 250 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, 45 mg, 40 mg, 35 mg, 30 mg, 25 mg, 20 mg, 15 mg, 10 mg, by mass of fluid in the at least one well. 63. A method according to any of claims 51 to 62, wherein the method is sensitive enough to detect changes of 5 mg, 4 mg, 3 mg, 2 mg, 1 mg, 500 micrograms ([mu] g), 300 [mu] g, 200 [mu] g or 100 [mu] g.   64. The method according to any of claims 51 to 63, further comprising detecting the temperature of at least one well.   65. The method of claim 64, further comprising detecting the temperature of the at least one well at at least two different time points.   66. The method of claim 64 or 65, further comprising adjusting the temperature of an individual well in response to the step of detecting the temperature.   67. A method according to any of claims 51 to 66, wherein at least one well in the multi-well plate is removable therefrom.