Using determination of different pitch heights

文档序号：1343386 发布日期：2020-07-17 浏览：4次中文

阅读说明：本技术 采用不同间距高度的测定 (Using determination of different pitch heights ) 是由斯蒂芬·Y·周丁惟戚骥张玙璠于 2018-02-09 设计创作，主要内容包括：本发明尤其提供了用于测定目的的操纵样品的装置和方法。(The present invention provides, inter alia, apparatus and methods for manipulating samples for assay purposes.)

1. A device for analyzing a liquid sample, comprising:

a first plate, a second plate, and a spacer, wherein:

i. the plates are movable relative to each other into different configurations;

one or both plates are flexible;

the first plate has first and second sample contacting areas at different locations on its inner surface, and the first plate has first and second thicknesses at the first and second sample contacting areas, respectively, wherein the sample contacting areas are for contacting a sample suspected of containing a target analyte, and wherein the first thickness is different from the second thickness;

said spacer being fixed to said first plate inner surface and having a predetermined substantially uniform height in each sample contacting area;

v. each of said spacers having a tip therein, and the tips of said spacers being substantially aligned in one surface;

wherein one of the configurations is an open configuration in which two plates are partially or fully separated, the spacing between the plates is not adjusted by spacers, and the sample is deposited on one or both of the plates; and is

Wherein another of the configurations is a closed configuration configured after deposition of the sample in the open configuration and in which at least a portion of the deposited sample is compressed by the two plates into a layer defined by the two plates and has a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact area and is accommodated by the plates and spacers in the respective sample contact area.

2. A method of analyzing a sample using a QMAX device, the method comprising:

(a) obtaining a sample suspected of containing a target analyte;

(b) obtaining a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

the first plate has first and second sample contacting areas on its inner surface at different locations, and the second plate has first and second sample contacting areas on its inner surface at different locations corresponding to the first and second sample contacting areas of the first plate, respectively, wherein the sample contacting areas are for contacting a sample;

the spacers are fixed to respective inner surfaces of one or both of the plates and have a predetermined substantially uniform height in each sample contact area;

v. the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

wherein the open configuration is one in which the two plates are partially or completely separated, the spacing between the two plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

wherein the pressing comprises conforming pressing a region of at least one of the plates in parallel or sequentially to press the plates together into the closed configuration, wherein the conforming pressing produces a substantially uniform pressure on the plates over at least a portion of the sample, and the pressing spreads at least a portion of the sample laterally between inner surfaces of the plates; and

wherein the closed configuration is configured after deposition of the sample in the open configuration, and in the closed configuration: the respective corresponding sample contact areas being on top of each other and at least a portion of the deposited sample being compressed by the two plates into a layer defined by the two plates and having a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas;

and

(e) analyzing the uniform thickness layer for a target analyte when the plate is in the closed configuration.

3. A method using a QMAX apparatus, said analyzing step (e) comprising, for performing a parallel multiplex assay:

(1) incubating the sample for a relevant length of time and then stopping the incubation;

(2) incubating the sample for a time equal to or longer than the minimum value of the length of correlation time and then assessing the binding of each analyte of interest to the binding site for a period of time equal to or less than the maximum value of the length of correlation time,

thereby generating a reaction in which a majority of the target analyte in the capture agent-target analyte-detection agent sandwich bound to each binding site is from the respective relevant volume of the sample at the end of the incubation in (1) or during the evaluation in (2);

wherein the incubation allows each analyte of interest to bind to a binding site and a detection agent, wherein the respective volume of interest is a portion of the sample above the respective storage site in the closed configuration.

4. The device of claim 1, wherein the uniform height of the spacers is in the range of 0.5 μ ι η to 100 μ ι η.

5. The device of any preceding claim, wherein the uniform height of the spacers is in the range of 0.5 μ ι η to 20 μ ι η.

6. The device of any preceding claim, wherein the difference between the uniform heights of the spacers in the first and second sample contact areas is in the range 0.5 to 100 μm.

7. The device of any preceding claim, wherein the difference between the uniform heights of the spacers in the first and second sample contact areas is in the range of 0.5 to 50 μm.

8. The device of any preceding claim, wherein the spacer has a predetermined substantially constant spacer pitch in each sample contact area.

9. The device of claim 8, wherein the constant spacer pitch of the spacers is in the range of 7 to 200 μm.

10. The device of claim 8, wherein the constant spacer pitch of the spacers is in the range of 50 to 150 μm.

11. A device as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 20 μm to 1 mm.

12. A device as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 100 μm to 500 μm.

13. The device of any preceding claim, wherein the average value of the uniform thickness of the layer in each sample contact area is substantially the same as the uniform height of the spacers therein, varying by less than 10%.

14. A device as claimed in any preceding claim, wherein the first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and containing a capture agent capable of binding and immobilising the target analyte.

15. The device of claim 14, wherein, in the closed configuration, the uniform thickness of the layer in any one of the sample contacting regions is substantially less than the predetermined lateral area of the binding sites therein.

16. The device of any preceding claim, wherein the second plate has first and second sample contacting areas at different locations corresponding to first and second sample contacting areas, respectively, of the first plate, wherein the corresponding sample contacting areas overlap each other in the closed configuration.

17. The device of claim 16, wherein the second plate has a storage site on at least one of the sample contact areas, the storage site having a predetermined lateral area and containing a detection agent that dissolves and diffuses in the sample when contacting the sample.

18. The device of any preceding claim, wherein the minimum separation between the edges of adjacent sample contact regions is substantially greater than the distance over which a target analyte or detection agent can diffuse for a relevant time, wherein the relevant time is:

i. about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform thickness layer in the closed configuration; and

shorter than the time it takes for the target analyte to diffuse laterally across the linear dimension of the predetermined region of the binding site.

19. The device of any preceding claim, wherein there is no fluidic isolation between adjacent sample contacting regions.

20. A method for analyzing a liquid sample, comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) a first plate, a second plate, and a spacer, wherein:

i. the plates are movable relative to each other into different configurations;

one or both plates are flexible;

said spacer being fixed to said first plate inner surface and having a predetermined substantially uniform height in each sample contacting area;

v. each of the spacers has a tip, and the tips of the spacers are substantially aligned in one surface;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

wherein the closed configuration is configured after deposition of the sample in the open configuration and in the closed configuration at least a portion of the deposited sample is compressed by the two plates into a layer confined by the two plates and having a respective substantially uniform thickness over each sample contact area, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas;

and

(e) analyzing the uniform thickness layer for a target analyte when the plate is in the closed configuration.

21. The method of claim 20, wherein the uniform height of the spacers is in the range of 0.5 to 100 μ ι η.

22. The method of any preceding claim, wherein the uniform height of the spacers is in the range of 0.5 to 20 μ ι η.

23. The method of any preceding claim, wherein the difference between the uniform heights of the spacers in different sample contact areas is in the range of 0.5 to 100 μm.

24. The method of any preceding claim, wherein the difference between the uniform heights of the spacers in different sample contact areas is in the range 0.5 μ ι η to 50 μ ι η.

25. The method of any preceding claim, wherein the spacer has a predetermined substantially constant spacer pitch in each sample contact area;

26. the method of claim 25, wherein the constant spacer pitch of the spacers is in the range of 7 to 200 μ ι η.

27. The method of claim 25, wherein the constant spacer pitch of the spacers is in the range of 50 to 150 μ ι η.

28. A method as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 20 to 1 mm.

29. A method as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 100 to 500 μm.

30. A method as claimed in any preceding claim, wherein the average value of the uniform thickness of the layer in each sample contact area is substantially the same as the uniform height of the spacers therein, varying by less than 10%.

31. The method of any preceding claim, further comprising: after step (d) and before step (e), removing the conformal compressive force, wherein a thickness of the uniform-thickness layer after removing the conformal compressive force is: (i) substantially the same as the uniform thickness layer prior to removal of the conformal compressive force, and (ii) less than 10% offset from the spacer height.

32. The method of any preceding claim, wherein the conformal compression is performed by a human hand.

33. The method of any preceding claim, wherein the conformal compression is provided by a pressurized liquid, a pressurized gas, or a conformal material.

34. The method of any preceding claim, wherein the sample deposition of step (c) is deposition directly from a subject to the plate without the use of any transfer device.

35. A method as claimed in any preceding claim, wherein during the deposition of step (c) the amount of sample deposited on the plate is unknown.

36. A method as claimed in any preceding claim, wherein the analysis in step (e) comprises performing the determination in the layer of uniform thickness.

37. The method of claim 36, wherein the assay is a binding assay or a biochemical assay.

38. The method of any preceding claim, wherein the first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and containing a capture agent capable of binding and immobilizing the target analyte.

39. A method according to claim 38, wherein in the closed configuration, the uniform thickness of the layer in any one of the sample contacting regions is substantially less than the predetermined lateral area of the binding sites therein.

40. The method of any preceding claim, wherein the second plate has a storage site on at least one of the sample contact regions, the storage site having a predetermined lateral area and containing a concentration of a detection agent that dissolves into and diffuses within the sample when contacting the sample.

41. The method of any preceding claim, wherein the minimum separation between the edges of adjacent sample contact regions is substantially greater than the distance of diffusion of target analyte or detection agent over a relevant time, wherein there is no fluidic isolation between the adjacent sample contact regions, wherein the relevant time lengths are:

i. about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

shorter than the time it takes for the target analyte to diffuse laterally across the linear dimension of the predetermined region of the binding site.

42. The method of any preceding claim, wherein the analysis of step (e) comprises:

(1) incubating the sample for a relevant length of time and then stopping the incubation;

(2) incubating the sample for a time equal to or longer than a minimum value of a correlation time length and then assessing binding of each analyte of interest to the binding site for a time period equal to or less than a maximum value of the correlation time length,

43. The method of any preceding claim, wherein the reaction is saturated in less than 60 seconds.

44. A method as claimed in any preceding claim, wherein the length of correlation time is in the range 60 seconds to 30 minutes.

45. The method of any preceding claim, wherein the analysis in step (e) comprises measuring a target analyte-related signal selected from the group consisting of:

i. luminescence selected from photoluminescence, electroluminescence and electrochemiluminescence;

light absorption, reflection, transmission, diffraction, scattering or diffusion;

(ii) surface Raman scattering,

an electrical impedance selected from the group consisting of resistance, capacitance, and inductance;

magnetic relaxation

Any combination of i-v.

Wherein the target analyte-related signal is a signal that is proportional to and reflective of the amount of target analyte in the sample.

46. The method of claim 45, wherein the analyzing step (e) comprises:

determining an optimal signal based on a target analyte-related signal measured from a sample-contact area of interest, wherein the sample-contact area of interest is a sample-contact area containing binding sites and/or storage sites for detecting the same target analyte.

47. The method of claim 45, wherein the optimal target analyte related signal is determined by selecting the measured target analyte related signal in a range between minimum and maximum detection thresholds for the plate and detectors for signal measurement.

48. The method of claim 45, wherein the optimal target analyte-related signal is determined by selecting a measured target analyte-related signal within the determined linear detection range, wherein the linear detection range is an intensity range of the target analyte-related signal, wherein the signal intensity is linearly related to the amount of the determined target analyte.

49. A method according to any preceding claim, wherein the analysing step (e) comprises calculating the volume of the sample volume of interest by measuring a lateral area of the sample volume of interest and calculating the volume from the lateral area and the predetermined spacer height, wherein the sample volume of interest is part or all of the volume of the sample.

50. The method of any preceding claim, wherein the analyzing step (e) comprises reading, image analysis or counting of the target analytes, or a combination thereof.

51. The method of any preceding claim, further comprising one or more washes.

52. The method of any preceding claim, wherein the deposited sample has a total volume of less than 0.5 μ L.

53. The method of any preceding claim, wherein the liquid sample is made from a biological sample selected from the group consisting of: amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma, or serum), breast milk, cerebrospinal fluid (CSF), cerumen (cerumen), chyle, chyme, endolymph, perilymph, stool, breath, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, saliva, exhaled breath condensate, sebum, semen, sputum, sweat, synovial fluid, tears, vomit, urine, and any combination thereof.

54. The method of any preceding claim, wherein the sample is an environmental liquid sample from a source selected from the group consisting of: a river, lake, pond, sea, glacier, iceberg, rain, snow, sewage, reservoir, tap or potable water, a solid sample from soil, compost, sand, rock, concrete, wood, brick, sewage, and any combination thereof.

55. The method of any preceding claim, wherein the sample is an ambient gaseous sample from a source selected from the group consisting of: air, subsea heat rejection, industrial exhaust, vehicle exhaust, and any combination thereof.

56. The method of any preceding claim, wherein the sample is a food sample selected from the group consisting of: raw materials, cooked foods, plant and animal food sources, pre-processed foods, partially or fully processed foods, and any combination thereof.

57. The method of any preceding claim, wherein the sample is human blood and the depositing step comprises: (a) piercing the skin of a person to release a drop of blood onto the skin; and (b) contacting the drop of blood with a filter without the use of a blood transfer means.

58. A method for parallel multiplexed assay of a liquid sample comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

the first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and comprising a capture agent capable of binding and immobilizing the target analyte;

v. the second plate has a storage site on at least one of the sample contacting regions, the storage site having a predetermined lateral area and containing a detection agent that dissolves and diffuses in the sample when contacting the sample;

said spacers being fixed to respective inner surfaces of one or both of said plates and having a predetermined substantially uniform height in each sample contact area; and

each of the spacers has a tip, and the tips of the spacers are substantially aligned in one surface;

wherein each capture agent, target analyte and corresponding detection agent is capable of forming a capture agent-target analyte-detection agent sandwich in the binding sites of the first plate;

wherein the open configuration is one in which the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacers, and the sample is deposited on one or both of the plates;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

wherein the closed configuration is configured after deposition of the sample in the open configuration, and in the closed configuration: the corresponding sample contact areas on the two plates are respectively on top of each other, and at least a portion of the deposited sample is compressed by the two plates into a layer defined by the two plates and has a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas;

and

(e) after (d) and when the panels are in the closed configuration:

(1) incubating the sample for a relevant length of time and then stopping the incubation;

wherein the incubation allows each target analyte to bind to a binding site and a detection agent, wherein the respective relevant volume is a portion of the sample above the respective storage site in the closed configuration, and

wherein the correlation time is:

(i) about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

(ii) shorter than the time it takes for the target analyte to diffuse laterally across the linear dimensions of the predetermined region of the binding site.

59. The method of claim 58, wherein step (e) comprises measuring a target analyte-related signal selected from the group consisting of:

i. luminescence selected from photoluminescence, electroluminescence and electrochemiluminescence;

light absorption, reflection, transmission, diffraction, scattering or diffusion;

(ii) surface Raman scattering,

an electrical impedance selected from the group consisting of resistance, capacitance, and inductance;

magnetic relaxation

Any combination of i-v.

Wherein the target analyte-related signal is a signal that is proportional to the binding of the target analyte to the binding site and that reflects the binding of the target analyte to the binding site.

60. The method of claim 59, wherein the analyzing step (e) comprises:

the optimal signal is determined from the target analyte-related signal measured from the sample-contact area of interest, which is the sample-contact area containing the binding sites and/or storage sites for detecting the same target analyte.

61. The method of claim 60, wherein the optimal target analyte related signal is determined by selecting the measured target analyte related signal in a range between minimum and maximum detection thresholds, wherein the minimum and maximum detection thresholds and detectors of the panel are used for signal measurement.

62. The method of claim 60, wherein the optimal target analyte-related signal is determined by selecting a measured target analyte-related signal within the determined linear detection range, wherein the linear detection range is an intensity range of the target analyte-related signal, wherein the signal intensity is linearly related to the amount of the determined target analyte.

63. The device or method of any preceding claim, wherein the binding site is defined by a patch of dried reagent.

64. The device or method of any preceding claim, wherein the binding site is between a pair of electrodes.

65. The device or method of any preceding claim, wherein one or both plate interior surfaces comprise one or more amplification sites that are each capable of amplifying the target analyte-related signal when the target analyte is within 500nm of an amplification site.

66. The apparatus or method of any preceding claim, wherein the plate has a thickness of less than 200 μ ι η.

67. The apparatus or method of any preceding claim, wherein the plate has a thickness of less than 100 μ ι η.

68. An apparatus or method as claimed in any preceding claim, wherein each of the plates has an area of less than 5cm²。

69. An apparatus or method as claimed in any preceding claim, wherein in the plateEach having an area of less than 2cm²。

70. The apparatus or method of any preceding claim, wherein at least one of the plates is partially or fully transparent.

71. A method as claimed in any preceding claim, wherein at least one of the plates is made of a flexible polymer.

72. An apparatus or method as claimed in any preceding claim, wherein at least one of the plates is a flexible plate and the thickness of the flexible plate times the young's modulus of the flexible plate is in the range 60 to 75GPa- μm.

73. The device or method of any preceding claim, wherein the spacer is a spacer having a cross-sectional shape selected from a circle, polygon, perfect circle, square, rectangle, oval, ellipse, or any combination thereof.

74. The apparatus or method of any preceding claim, wherein the spacers have a cylindrical shape and a substantially flat top surface, wherein for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.

75. The device or method of any preceding claim, wherein each spacer has a ratio of a lateral dimension of the spacer to a height thereof of at least 1.

76. The device or method of any preceding claim, wherein the smallest lateral dimension of the spacer is less than or substantially equal to the smallest dimension of a target analyte in the sample.

77. The apparatus or method of any preceding claim, wherein the spacer has a cylindrical shape and the sidewall corners of the spacer have a rounded shape with a radius of curvature of at least 1 μ ι η.

78. The apparatus or method of any preceding claim, wherein the spacer has at least 100/mm²The density of (c).

79. The apparatus or method of any preceding claim, wherein the spacer has at least 1000/mm²The density of (c).

80. The apparatus or method of any preceding claim, wherein the spacer has a fill factor of at least 1%, wherein the fill factor is a ratio of the spacer area in contact with the uniform thickness layer to the total plate area in contact with the uniform thickness layer.

81. The apparatus or method of any preceding claim, wherein the young's modulus of the spacer multiplied by the fill factor of the spacer is equal to or greater than 10MPa, wherein the fill factor is the ratio of the spacer area in contact with the uniform thickness layer to the total plate area in contact with the uniform thickness layer.

82. An apparatus or method as claimed in any preceding claim, wherein

a. At least one of the plates is flexible, and

b. for the flex, the fourth power of the spacer spacing (ISD) divided by the thickness (h) of the flex and the Young's modulus (E) of the flex⁴/(hE), equal to or less than 106 μm³/GPa。

83. The apparatus or method of any preceding claim, wherein the spacer is fixed to a plate by directly stamping the plate or injection moulding the plate.

84. The device or method of any preceding claim, wherein the material of the plate and the spacer is independently selected from polystyrene, PMMG, PC, COC, COP or other plastics.

85. A method comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) obtaining a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

said spacer being fixed to said first plate inner surface and having a predetermined substantially uniform height in each sample contacting area;

v. the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

in the closed configuration: the respective corresponding sample contact areas being on top of each other and at least a portion of the deposited sample being compressed by the two plates into a layer defined by the two plates and having a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas;

and

(e) analyzing the presence or absence, amount and/or concentration of the target analyte in the uniform thickness layer by: (1) measuring a target analyte-related signal in a layer over the first and second sample contact areas, respectively; (2) determining a ratio of a signal measured on the first sample contact area to a signal measured on a second sample contact area, wherein the target analyte-related signal is a signal that is proportional to and reflective of the amount of target analyte in the sample.

86. A device for analyzing a liquid sample, comprising:

a first plate, a second plate, and a spacer, wherein:

i. the plates are movable relative to each other into different configurations;

one or both plates are flexible;

the spacers are fixed to respective inner surfaces of one or both of the plates and have a predetermined substantially uniform height in each sample contact area; and

v. the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

wherein one of the configurations is an open configuration in which the two plates are partially or fully separated, the spacing between the plates is not adjusted by spacers, and the sample is deposited on one or both of the plates; and is

Wherein another of the configurations is a closed configuration, the closed configuration being configured after deposition of the sample in the open configuration, and in the closed configuration: the respective corresponding sample contact areas are on top of each other and at least a portion of the deposited sample is compressed by the two plates into a layer defined by the two plates and has a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas.

87. An apparatus for parallel multiplexed assays of a liquid sample, comprising:

a first plate, a second plate, and a spacer, wherein:

i. the plates are movable relative to each other into different configurations;

one or both of the plates are flexible;

v. the second plate has first and second sample contacting areas with a storage site on at least one of the sample contacting areas, the storage site having a predetermined lateral area and containing a detection agent that dissolves and diffuses in the sample when contacting the sample;

said spacers being fixed to respective inner surfaces of one or more of said plates and having a predetermined substantially uniform height in each sample contact area; and

the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

wherein another of the configurations is a closed configuration, the closed configuration being configured after deposition of the sample in the open configuration, and in the closed configuration: the respective corresponding sample contact areas on the two plates are on top of each other and at least a portion of the deposited sample is compressed by the two plates into a layer defined by the two plates and has a respective substantially uniform thickness over each of the sample contact areas, wherein the uniform thickness of the layer is defined by the respective sample contact areas of the plates and is accommodated by the plates and spacers in the respective sample contact areas;

wherein each capture agent, target analyte and corresponding detection agent is capable of forming a capture agent-target analyte-detection agent sandwich in the binding sites of the first plate;

wherein there is no fluidic isolation between adjacent sample contact regions; and

wherein the minimum separation between edges of adjacent sample contact regions is substantially greater than the distance that the target analyte or detection agent diffuses over a time period of:

(1) about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

(2) shorter than the time it takes for the target analyte to diffuse laterally across the linear dimensions of the predetermined region of the binding site.

88. A method for analyzing a liquid sample, comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) obtaining a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

the spacers are fixed to respective inner surfaces of one or both of the plates and have a predetermined substantially uniform height in each sample contact area; and

v. the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

and

(e) analyzing the target analyte in the uniform thickness layer when the plate is in the closed configuration.

89. A method for analyzing a liquid sample, comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) obtaining a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

the spacers are fixed to respective inner surfaces of one or both of the plates and have a predetermined substantially uniform height in each sample contact area; and

v. the height of the spacer in the first sample contact area is different from its height in the second sample contact area;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

and

90. A method for parallel multiplexed assay of a liquid sample comprising the steps of:

(a) obtaining a sample suspected of containing a target analyte;

(b) a first plate, a second plate, and a spacer, wherein:

i. the panels are movable relative to one another into different configurations, including an open configuration and a closed configuration;

one or both of the plates are flexible;

The first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and comprising a capture agent capable of binding and immobilizing the target analyte.

said spacers being fixed to respective inner surfaces of one or both of said plates and having a predetermined substantially uniform height in each sample contact area; and

wherein each capture agent, target analyte and corresponding detection agent is capable of forming a capture agent-target analyte-detection agent sandwich in the binding sites of the first plate;

(d) after (c), bonding the two panels together and pressing the panels into the closed configuration,

and

(e) after (d) and when the panels are in the closed configuration:

(1) incubating the sample for a relevant length of time and then stopping the incubation;

wherein the correlation time is:

(i) about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

(ii) shorter than the time it takes for the target analyte to diffuse laterally across the linear dimensions of the predetermined region of the binding site.

91. The apparatus of claim 86, wherein the uniform height of the spacers is in the range of 0.5 to 100 μm.

92. The device of any preceding claim, wherein the uniform height of the spacers is in the range of 0.5 to 20 μm.

93. The device of any preceding claim, wherein the difference between the uniform heights of the spacers in the first and second sample contact areas is in the range 0.5 to 100 μm.

94. The device of any preceding claim, wherein the difference between the uniform heights of the spacers in the first and second sample contact areas is in the range 0.5 to 50 μm.

95. The device of any preceding claim, wherein the spacer has a predetermined substantially constant spacer pitch in each sample contact area.

96. The apparatus of claim 86, wherein the constant spacer pitch of the spacers is in the range of 7 to 200 μm.

97. The apparatus of claim 86, wherein the constant spacer pitch of the spacers is in the range of 50 to 150 μm.

98. A device as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 20 to 1 mm.

99. A device as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 100 to 500 μm.

100. The device of any preceding claim, wherein the average value of the uniform thickness of the layer in each sample contact area is substantially the same as the uniform height of the spacers therein, varying by less than 10%.

101. A device as claimed in any preceding claim, wherein the first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and containing a capture agent capable of binding and immobilising the target analyte.

102. The device of claim 101, wherein, in the closed configuration, the uniform thickness of the layer in any one of the sample contacting regions is substantially less than the predetermined lateral area of the binding sites therein.

103. The device of any preceding claim, wherein the second plate has first and second sample contacting areas at different locations corresponding to first and second sample contacting areas, respectively, of the first plate, wherein the corresponding sample contacting areas overlap each other in the closed configuration.

104. The device of claim 103, wherein the second plate has a storage site on at least one of the sample contact regions, the storage site having a predetermined lateral area and containing a detection agent that dissolves and diffuses in the sample when contacting the sample.

105. The device of any preceding claim, wherein the minimum separation between the edges of adjacent sample contact regions is substantially greater than the distance over which a target analyte or detection agent can diffuse for a relevant time, wherein the relevant time is:

about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

shorter than the time it takes for the target analyte to diffuse laterally across the linear dimension of the predetermined region of the binding site.

106. The device of any preceding claim, wherein there is no fluidic isolation between adjacent sample contacting regions.

107. The method of any one of claim 88 or claim 89, wherein the uniform height of the spacers is in the range of 0.5 to 100 μm.

108. The method of any preceding claim, wherein the uniform height of the spacers is in the range of 0.5 to 20 μ ι η.

109. The method of any preceding claim, wherein the difference between the uniform heights of the spacers in different sample contact areas is in the range of 0.5 to 100 μm.

110. The method of any preceding claim, wherein the difference between the uniform heights of the spacers in different sample contact areas is in the range of 0.5 to 50 μm.

111. The method of any preceding claim, wherein the spacer has a predetermined substantially constant spacer pitch in each sample contact area;

112. the method of claim 111, wherein the constant spacer pitch of the spacers is in the range of 7 to 200 μ ι η.

113. The method of claim 111, wherein the constant spacer pitch of the spacers is in the range of 50 to 150 μ ι η.

114. A method as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 20 to 1 mm.

115. A method as claimed in any preceding claim, wherein the spacing between the edges of adjacent sample contact regions is in the range 100 to 500 μm.

116. A method as claimed in any preceding claim, wherein the average value of the uniform thickness of the layer in each sample contact area is substantially the same as the uniform height of the spacers therein, varying by less than 10%.

117. The method of any preceding claim, further comprising: after step (d) and before step (e), removing the conformal compressive force, wherein a thickness of the uniform-thickness layer after removing the conformal compressive force is: (i) substantially the same as the uniform thickness layer prior to removal of the conformal compressive force, and (ii) less than 10% offset from the spacer height.

118. The method of any preceding claim, wherein the conformal compression is performed by a human hand.

119. The method of any preceding claim, wherein the conformal compression is provided by a pressurized liquid, a pressurized gas, or a conformal material.

120. The method of any preceding claim, wherein the sample deposition of step (c) is deposition directly from a subject to the plate without the use of any transfer device.

121. A method as claimed in any preceding claim, wherein during the deposition of step (c) the amount of sample deposited on the plate is unknown.

122. A method as claimed in any preceding claim, wherein the analysis in step (e) comprises performing the determination in the layer of uniform thickness.

123. The method of claim 122, wherein the assay is a binding assay or a biochemical assay.

124. The method of any preceding claim, wherein the first plate has a binding site on at least one of the sample contact regions, the binding site having a predetermined lateral area and containing a capture agent capable of binding and immobilizing the target analyte.

125. The method of claim 124, wherein in the closed configuration, the uniform thickness of the layer in any of the sample contacting regions is substantially less than the predetermined lateral area of the binding sites therein.

126. The method of any preceding claim, wherein the second plate has a storage site on at least one of the sample contact regions, the storage site having a predetermined lateral area and containing a concentration of a detection agent that dissolves into and diffuses within the sample when contacting the sample.

127. The method of any preceding claim, wherein the minimum separation between the edges of adjacent sample contact regions is substantially greater than the distance of diffusion of target analyte or detection agent over a relevant time, wherein there is no fluidic isolation between the adjacent sample contact regions, wherein the relevant time lengths are:

about equal to or longer than the time it takes for the target analyte to diffuse through the thickness of the uniform-thickness layer in the closed configuration; and

shorter than the time it takes for the target analyte to diffuse laterally across the linear dimension of the predetermined region of the binding site.

128. The method of any preceding claim, wherein the analysis of step (e) comprises:

(1) incubating the sample for a relevant length of time and then stopping the incubation;

129. The method of any preceding claim, wherein the reaction is saturated in less than 60 seconds.

130. A method as claimed in any preceding claim, wherein the length of correlation time is in the range 60 seconds to 30 minutes.

131. The method of any preceding claim, wherein the analysis in step (e) comprises measuring a target analyte-related signal selected from the group consisting of:

luminescence selected from photoluminescence, electroluminescence and electrochemiluminescence;

light absorption, reflection, transmission, diffraction, scattering or diffusion;

surface raman scattering;

x. an electrical impedance selected from the group consisting of resistance, capacitance, and inductance;

magnetic relaxation; and

any combination of xi.

Wherein the target analyte-related signal is a signal that is proportional to and reflective of the amount of target analyte in the sample.

132. The method of claim 131, wherein the analyzing step (e) comprises:

133. The method of claim 131, wherein said optimal target analyte related signal is determined by selecting said measured target analyte related signal in a range between a minimum and a maximum detection threshold of said plate and a detector for signal measurement.

134. The method of claim 131, wherein the optimal target analyte-related signal is determined by selecting a measured target analyte-related signal within the determined linear detection range, wherein the linear detection range is an intensity range of the target analyte-related signal, wherein the signal intensity is linearly related to the amount of the determined target analyte.

135. A method according to any preceding claim, wherein the analysing step (e) comprises calculating the volume of the sample volume of interest by measuring a lateral area of the sample volume of interest and calculating the volume from the lateral area and the predetermined spacer height, wherein the volume of interest is part or all of the volume of the sample.

136. The method of any preceding claim, wherein the analyzing step (e) comprises reading, image analysis or counting of the target analytes, or a combination thereof.

137. The method of any preceding claim, further comprising one or more washes.

138. The method of any preceding claim, wherein the deposited sample has a total volume of less than 0.5 μ L.

139. The method of any preceding claim, wherein the liquid sample is made from a biological sample selected from the group consisting of: amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma, or serum), breast milk, cerebrospinal fluid (CSF), cerumen (cerumen), chyle, chyme, endolymph, perilymph, stool, breath, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, saliva, exhaled breath condensate, sebum, semen, sputum, sweat, synovial fluid, tears, vomit, urine, and any combination thereof.

140. The method of any preceding claim, wherein the sample is an environmental liquid sample from a source selected from the group consisting of: a river, lake, pond, sea, glacier, iceberg, rain, snow, sewage, reservoir, tap or potable water, a solid sample from soil, compost, sand, rock, concrete, wood, brick, sewage, and any combination thereof.

141. The method of any preceding claim, wherein the sample is an ambient gaseous sample from a source selected from the group consisting of: air, subsea heat rejection, industrial exhaust, vehicle exhaust, and any combination thereof.

142. The method of any preceding claim, wherein the sample is a food sample selected from the group consisting of: raw materials, cooked foods, plant and animal food sources, pre-processed foods, partially or fully processed foods, and any combination thereof.

143. The method of any preceding claim, wherein the sample is human blood and the depositing step comprises: (a) piercing the skin of a person to release a drop of blood onto the skin; and (b) contacting the drop of blood with a filter without the use of a blood transfer means.

144. The method of claim 90, wherein step (e) comprises measuring a target analyte-related signal selected from the group consisting of:

luminescence selected from photoluminescence, electroluminescence and electrochemiluminescence;

light absorption, reflection, transmission, diffraction, scattering or diffusion;

surface raman scattering;

x. an electrical impedance selected from the group consisting of resistance, capacitance, and inductance;

magnetic relaxation; and

any combination of xi.

145. The method of claim 144, wherein the analyzing step (e) comprises:

146. The method of claim 145, wherein the optimal target analyte related signal is determined by selecting the measured target analyte related signal in a range between a minimum and a maximum detection threshold of the panel and a detector for signal measurement.

147. The method of claim 145, wherein the optimal target analyte-related signal is determined by selecting a measured target analyte-related signal within a linear detection range of the assay, wherein the linear detection range is a range of intensities of the target analyte-related signal, wherein the signal intensities are linearly related to an amount of the determined target analyte.

148. The device or method of any preceding claim, wherein the binding site is defined by a patch of dried reagent.

149. The device or method of any preceding claim, wherein the binding site is between a pair of electrodes.

150. The device or method of any preceding claim, wherein one or both plate interior surfaces comprise one or more amplification sites that are each capable of amplifying the target analyte-related signal when the target analyte is within 500nm of an amplification site.

151. The apparatus or method of any preceding claim, wherein the plate has a thickness of less than 200 μ ι η.

152. The apparatus or method of any preceding claim, wherein the plate has a thickness of less than 100 μ ι η.

153. An apparatus or method as claimed in any preceding claim, wherein each of the plates has an area of less than 5cm²。

154. An apparatus or method as claimed in any preceding claim, wherein each of the plates has an area of less than 2cm²。

155. The apparatus or method of any preceding claim, wherein at least one of the plates is partially or fully transparent.

156. A method as claimed in any preceding claim, wherein at least one of the plates is made of a flexible polymer.

157. An apparatus or method as claimed in any preceding claim, wherein at least one of the plates is a flexible plate and the thickness of the flexible plate times the young's modulus of the flexible plate is in the range 60 to 75GPa- μm.

158. The device or method of any preceding claim, wherein the spacer is a spacer having a cross-sectional shape selected from a circle, polygon, perfect circle, square, rectangle, oval, ellipse, or any combination thereof.

159. The apparatus or method of any preceding claim, wherein the spacers have a cylindrical shape and a substantially flat top surface, wherein for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.

160. The device or method of any preceding claim, wherein each spacer has a ratio of a lateral dimension of the spacer to a height thereof of at least 1.

161. The device or method of any preceding claim, wherein the smallest lateral dimension of the spacer is less than or substantially equal to the smallest dimension of a target analyte in the sample.

162. The apparatus or method of any preceding claim, wherein the spacer has a cylindrical shape and the sidewall corners of the spacer have a rounded shape with a radius of curvature of at least 1 μ ι η.

163. The apparatus or method of any preceding claim, wherein the spacer has at least 100/mm²The density of (c).

164. The apparatus or method of any preceding claim, wherein the spacer has at least 1000/mm²The density of (c).

165. The apparatus or method of any preceding claim, wherein the spacer has a fill factor of at least 1%, wherein the fill factor is a ratio of the spacer area in contact with the uniform thickness layer to the total plate area in contact with the uniform thickness layer.

166. The apparatus or method of any preceding claim, wherein the young's modulus of the spacer multiplied by the fill factor of the spacer is equal to or greater than 10MPa, wherein the fill factor is the ratio of the spacer area in contact with the uniform thickness layer to the total plate area in contact with the uniform thickness layer.

167. An apparatus or method as claimed in any preceding claim, wherein

a. At least one of the plates is flexible, and

b. for the flex, the fourth power of the spacer spacing (ISD) divided by the thickness (h) of the flex and the Young's modulus (E) of the flex⁴/(hE), equal to or less than 106 μm³/GPa。

168. The apparatus or method of any preceding claim, wherein the spacer is fixed to a plate by directly stamping the plate or injection moulding the plate.

169. The device or method of any preceding claim, wherein the material of the plate and the spacer is independently selected from polystyrene, PMMG, PC, COC, COP or other plastics.

170. An apparatus for lysing a component in a liquid sample, comprising:

a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

each plate has a sample contacting area on its respective sample surface for contacting a sample, wherein the sample comprises at least a target lysis component.

One or both of the plates comprises a column fixed to the respective sample contacting area, and

the column has at least one lateral dimension of the top surface of the column that is less than 200% of the target lysis component,

wherein, in the open configuration, the two plates are partially or fully separated, the spacing between the plates is not adjusted by the column, and the sample is deposited on one or both of the plates;

wherein the closed configuration is configured after deposition of the sample in the open configuration and in the closed configuration the relevant volume of the sample is compressed by the two plates into a layer of very uniform thickness and the uniform thickness of the layer is limited by the sample contacting area of the plates and is accommodated by the plates and the columns;

wherein a substantial portion of a target lytic component of the sample in the volume of interest is lysed by the column during transition of the plate from the open configuration to the closed configuration; and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

171. A method of mechanically lysing a component in a liquid sample, comprising the steps of:

(a) having a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

each plate has a sample contacting area on its respective sample surface for contacting a sample, wherein the sample comprises at least a target lysis component.

One or both of the plates comprises a column fixed to the respective sample contacting area, and

the column has at least one lateral dimension of the top surface of the column that is less than 200% of the target lysis component,

(b) depositing the sample on one or both of the plates; when the plates are configured in the open configuration, wherein in the open configuration the two plates are partially or fully separated and the spacing between the plates is not adjusted by posts;

(c) bringing the two plates together and pressing the two plates against each other with force to enter the closed configuration, during which a substantial portion of the target lytic component of the sample in the relevant volume is lysed by the column;

wherein in the closed configuration the relevant volume of the sample is compressed by the two plates into a layer of very uniform thickness, and the uniform thickness of the layer is limited by the sample contact area of the plates and is regulated by the plates and the columns, and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

172. An apparatus for selectively lysing a target percentage of components in a liquid sample, comprising:

a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

each of the plates has a sample contacting area on its respective sample surface for contacting a sample, wherein the sample comprises at least a target lysis component, and

one or both of the plates comprising a column immobilised to a respective sample contacting region,

wherein at least one parameter of the column is selected such that a target percentage of a target lysis component of the sample in the volume of interest is lysed by the column during the process of transitioning the plate from the open configuration to the closed configuration; and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

173. A method of selectively lysing a target percentage of components in a liquid sample, comprising the steps of:

(a) having a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

each of the plates has a sample contacting area on its respective sample surface for contacting a sample, wherein the sample comprises at least a target lysis component, and

one or both of the plates comprising a column immobilised to a respective sample contacting region,

(c) bringing the two plates together and pressing the two plates against each other with force to enter the closed configuration,

wherein in the closed configuration the relevant volume of the sample is compressed by the two plates into a layer of very uniform thickness, and the uniform thickness of the layer is limited by the sample contacting area of the plates and is regulated by the plates and the columns,

Wherein the relevant volume of the sample is a portion or all of the volume of the sample.

174. An apparatus for lysing a component in a liquid sample, comprising:

a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

each plate has a sample contacting area on its respective sample surface for contacting a sample, wherein the sample comprises at least one target lysis component and at least one non-target lysis component, and

one or both of the plates comprising a column immobilised to a respective sample contacting region,

wherein at least one parameter of the column is selected such that during the process of transitioning the plate from the open configuration to the closed configuration, a substantial portion of the target lysis component of the sample in the volume of interest is lysed by the column and a substantial portion of the non-target lysis component in the volume of interest is not lysed; and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

175. A method of selectively lysing a component in a liquid sample, comprising the steps of:

(a) having a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

one or both of the plates comprising a column immobilised to a respective sample contacting region,

(c) bringing the two plates together and pressing the two plates against each other with force to enter the closed configuration,

wherein in the closed configuration the relevant volume of the sample is compressed by the two plates into a layer of very uniform thickness, and the uniform thickness of the layer is limited by the sample contacting area of the plates and is regulated by the plates and the columns,

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

176. An apparatus for lysing a component in a liquid sample, comprising:

a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

the first plate has a first sample contact area at one location and a second sample contact area at another location on its respective sample surface, wherein the sample contact areas are for contacting a sample, wherein the sample comprises at least one target lysis component, and

one or both of the plates comprising a column immobilised to a respective sample contacting region,

wherein at least one parameter of the column is selected such that (a) the column has at least one parameter that is substantially the same in the first sample contact region and a different at least one parameter that is substantially the same in the second sample contact region, and (b) a substantial portion of a target lysis component of a sample above the first sample contact region is lysed by the column while a target lysis component above the second sample contact region is not lysed during a transition of the plate from an open configuration to a closed configuration; and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

177. A method of selectively lysing a component in a liquid sample, comprising the steps of:

(a) having a first plate and a second plate, wherein:

i. the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration,

the first plate has a first sample contact area at one location and a second sample contact area at another location on its respective sample surface, wherein the sample contact area is for contacting a sample, wherein the sample comprises at least one target lysis component, and

one or both of the plates comprises a column immobilised to the respective sample contacting area,

(c) bringing the two plates together and pressing the two plates against each other with force to enter the closed configuration,

wherein in the closed configuration the relevant volume of the sample is compressed by the two plates into a layer of very uniform thickness, and the uniform thickness of the layer is limited by the sample contacting area of the plates and is regulated by the plates and the columns,

wherein at least one parameter of the column is selected such that (a) the column has substantially the same at least one parameter in the first sample contact region and a different substantially the same at least one parameter in the second sample contact region, and (b) a substantial portion of a target lysis component of a sample above the first sample contact region is lysed by the column while a target lysis component above the second sample contact region is not lysed during a transition of the plate from an open configuration to a closed configuration; and

wherein the relevant volume of the sample is a portion or all of the volume of the sample.

178. The apparatus or method of any preceding claim, wherein at least one of the plates is flexible.

179. The apparatus or method of any preceding claim, wherein the base of at least one of the posts has an approximately circular shape.

180. The apparatus or method of any preceding claim, wherein the base of at least one of the posts has an approximate shape selected from the group consisting of: polygonal, pyramidal, elliptical, and elongated strips, as well as combinations thereof with circular shapes, and any combination thereof.

181. The device or method of any preceding claim, wherein the base of at least one of the pillars has a lateral dimension of 1nm or less, 10nm or less, 50nm or less, 100nm or less, 200nm or less, 500nm or less, 800nm or less, 1000nm or less, 2000nm or less, 3000nm or less, 5000nm or less, 10 microns or less, 20 microns or less, 30 microns or less, 50 microns or less, 100 microns or less, 150 microns or less, 200 microns or less, 300 microns or less, 500 microns or less, 800 microns or less, 1mm or less, 2mm or less, 4mm or less, or within a range between any two values.

182. The apparatus or method of any preceding claim, wherein the top of at least one of the posts has an approximately circular shape.

183. The apparatus or method of any preceding claim, wherein the top of at least one of the pillars has an approximate shape selected from the group consisting of: polygonal, pyramidal, elliptical, and elongated strips, as well as combinations thereof with circular shapes, and any combination thereof.

184. The device or method of any preceding claim, wherein the top of at least one of the pillars has a lateral dimension of 1nm or less, 10nm or less, 50nm or less, 100nm or less, 200nm or less, 500nm or less, 800nm or less, 1000nm or less, 2000nm or less, 3000nm or less, 5000nm or less, 10 microns or less, 20 microns or less, 30 microns or less, 50 microns or less, 100 microns or less, 150 microns or less, 200 microns or less, 300 microns or less, 500 microns or less, 800 microns or less, 1mm or less, 2mm or less, 4mm or less, or a range between any two values.

185. The apparatus or method of any preceding claim, wherein the spacing between two nearest pillars of the plurality of elements is in the range 2nm to less than 500 microns.

186. The device or method of any preceding claim, wherein the sample comprises whole blood or fractionated blood.

187. The device or method of any preceding claim, wherein the sample comprises any one or combination of the group consisting of: amniotic fluid, aqueous humor, vitreous humor, breast milk, cerebrospinal fluid (CSF), cerumen (cerumen), chyle, chyme, endolymph, perilymph, stool, gastric acid, gastric juice, lymph fluid, mucus, nasal drainage fluid, sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, sebum, semen, saliva, sweat, synovial fluid, tears, vomit, and exhaled condensate.

188. The method of any preceding claim, comprising converting a sample containing at least one target lysis component from a non-liquid phase to a liquid phase prior to step (i).

189. The device or method of any preceding claim, wherein at least one target lysis component in the sample is spread over at least one of the two plates on which the cell sample is deposited.

190. The apparatus or method of any preceding claim, wherein the target percentage is 5% or less, 10% or less, 20% or less, 30% or less, 40% or less, 50% or less, 60% or less, 70% or less, 80% or less, 90% or less, 95% or less, 98% or less, 99% or less, 100% or less, or any value within a range between any two of these numbers.

191. The apparatus or method of any preceding claim, wherein a substantial portion is at least 51%, 60%, 70%, 80%, 90%, 95% or 99%.

192. An apparatus or method as claimed in any preceding claim, wherein the substantially uniform layer is ofArea equal to or greater than 0.1mm²、0.5mm²、1mm²、3mm²、5mm²、10mm²、20mm²、50mm²、70mm²、100mm²、200mm²、500mm²、800mm²、1000mm²、2000mm²、5000mm²、10000mm²、20000mm²、50000mm²Or 100000mm²(ii) a Or within a range between any two values.

193. A device or method as claimed in any preceding claim, wherein the respective plate further comprises a reagent layer on one or both of the sample contact regions.

194. The device or method of any preceding claim, wherein the area of the first sample contacting region or the second sample contacting region is equal to or greater than 0.1mm²、0.5mm²、1mm²、3mm²、5mm²、10mm²、20mm²、50mm²、70mm²、100mm²、200mm²、500mm²、800mm²、1000mm²、2000mm²、5000mm²、10000mm²、20000mm²、50000mm²Or 100000mm²(ii) a Or within a range between any two values.

195. The method of any preceding claim, wherein the compressive force is applied by a human hand.

196. The method of any preceding claim, wherein the compressive force is applied by gas or fluid pressure.

197. The apparatus or method of any preceding claim, wherein the column has:

i. a shape of a pillar having a substantially uniform cross-section and a flat top surface;

a width to height ratio equal to or greater than 1;

a fill factor equal to or greater than 1%; and

the spacer has a fill factor multiplied by a Young's modulus equal to 2MPa or greater;

wherein the fill factor is the ratio of spacer contact area to total plate area;

198. the apparatus or method of any preceding claim, wherein the average value of the uniform thickness of the layer is substantially the same as the uniform height of the spacer, varying by less than 10%.

199. The device or method of any preceding claim, wherein the sample further comprises a second target lysis component.

200. The device or method of any preceding claim, wherein in the closed configuration at least 90% of the target lytic components are lysed and at least 90% of the non-target lytic components are lysed.

201. The device or method of any preceding claim, wherein in the closed configuration at least 99% of the target lytic components are lysed and at least 99% of the non-target lytic components are lysed.

202. The device or method of any preceding claim, wherein in the closed configuration, at least 90% of all targeted lysis components are lysed and at least 90% of non-targeted lysis components are lysed.

203. The device or method of any preceding claim, wherein the uniform thickness layer varies by less than 30 nm.

204. The apparatus or method of any preceding claim, wherein the thickness-uniform sample layer has a thickness uniformity of up to +/-5%.

205. The apparatus or method of any preceding claim, wherein the post has a cross-sectional shape selected from circular, polygonal, perfect circular, square, rectangular, oval, elliptical, or any combination thereof.

206. The device or method of any preceding claim, wherein analyzing the non-target components comprises counting the number of the non-lysed target analytes and calculating the concentration of the non-target components.

207. The apparatus or method of any preceding claim, wherein the column has:

i. a shape of a pillar having a substantially uniform cross-section and a flat top surface;

a width to height ratio equal to or greater than 1;

a predetermined constant spacer pitch in the range of 10 μ ι η to 200 μ ι η;

a fill factor equal to or greater than 1%; and

v. the filling factor of the spacer multiplied by the young's modulus equals 2MPa or more;

wherein the fill factor is a ratio of the spacer contact area to a total plate area;

208. the device or method of any preceding claim, wherein pressing the panels into the closed configuration is done in parallel or sequentially, the parallel pressing simultaneously applying an external force on the intended area, and the sequential pressing applying an external force on a portion of the intended area and gradually moving to other areas.

209. The device or method of any preceding claim, wherein the blood sample is stained prior to analysis.

210. The device or method of any preceding claim, wherein the blood sample is stained with Acridine Orange (AO).

211. The device or method of any preceding claim, wherein a staining reagent is coated on at least one sample contacting area and the blood sample is stained with the staining reagent.

212. The device or method of any preceding claim, wherein the blood sample is analysed by:

i. illuminating at least a portion of the blood sample in the thickness-uniforming layer;

acquiring one or more images of the cells using a CCD or CMOS sensor;

identifying platelets in the image using a computer; and

counting the number of platelets in the image region.

213. The apparatus or method of any preceding claim, wherein the thickness-uniform sample layer has a thickness uniformity of up to +/-5%.

214. The device or method of any preceding claim, wherein one or both of the plates comprises a detection agent on the respective sample contact region capable of selectively binding a target analyte in the sample.

215. The device or method of any preceding claim, wherein one or both of the plates comprises a capture agent on the respective sample contacting area capable of selectively binding and immobilizing a target analyte in the sample.

216. The method of any preceding claim, further comprising: incubating the assay of the sample when the two plates are in the closed configuration.

Technical Field

In particular, the present invention relates to devices and methods for performing biological and chemical analyses.

Background

In biological and chemical analysis, new methods are needed to manipulate samples. The present invention provides, inter alia, apparatus and methods for manipulating samples for assay purposes.

Disclosure of Invention

The following summary is not intended to include all features and aspects of the present invention.

One aspect of the present invention is to provide an apparatus that allows a sample sandwiched between two plates to have different thicknesses in different regions of the plates.

Another aspect of the invention is to control the spacing (i.e., gap) between the two plates to selectively lyse only certain types of cells and not others.

Another aspect of the invention is the use of spacers to control the spacing (i.e., gap) between the two plates and to control the spacing (and thus the sample thickness) accurately with sub-micron accuracy and uniformly over a large area.

Another aspect of the invention is the use of spacers to sandwich the sample between the two plates to have different thicknesses in different regions of the plates.

Another aspect of the invention is the use of different sample thicknesses in different regions of the plate sandwiching the sample to perform certain assay functions, including but not limited to: (i) selective lysis of cells in different regions, (ii) different amounts of analyte in the sample in different regions, (iii) reduction of Hook effect, etc.

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