MICROWELL ARRAY ARTICLES AND METHODS OF USE
The disclosure provides microstructured articles and methods useful for detecting an analyte in a sample. The articles include microwell arrays. The articles can be used with an optical system component in methods to detect or characterize an analyte
1 . An article, comprising
a microstructured layer with upper and lower major surfaces, comprising a plurality of optically-isolated microwells extending between the upper and lower major surfaces; and
an optically-transmissive flexible layer coupled to the lower major surface of the microstructured layer;
wherein each microwell in the microstructured layer comprises a top opening, a bottom opening, and at least one side wall extending between the top opening and the bottom opening;
wherein the optically-transmissive flexible layer has an average thickness of about 2 μm to about 50 μm.
2 . An article, comprising
a microstructured layer with upper and lower major surfaces, comprising a plurality of optically-isolated microwells extending below the upper major surface; and
an optically-transmissive flexible layer coupled to the lower major surface of the microstructured layer;
wherein each microwell in the microstructured layer comprises an opening, an optically-transmissive bottom wall, and at least one side wall extending between the opening and the bottom wall;
wherein the bottom wall has a thickness of about 0.1 μm to about 5 μm.
3 . An article, comprising
a microstructured layer with upper and lower major surfaces, comprising a plurality of optically-isolated microwells extending below the upper major surface; and
an optically-transmissive flexible layer coupled to the lower major surface of the microstructured layer;
wherein each microwell in the microstructured layer comprises an opening, an optically-transmissive bottom wall, and at least one side wall extending between the opening and the bottom wall;
wherein a thickness (t) is defined by a thickness of the bottom wall plus a thickness of the optically-transmissive flexible layer;
wherein t is about 2 μm to about 55 μm.
4 . The article of claim 1 , wherein the microstructured layer comprises a colorant.
5 . The article of claim 4 , wherein the colorant is selected from the group consisting of carbon black, fuchsin, carbazole violet, and Foron Brilliant Blue.
6 . The article of claim 1 , wherein the optically-transmissive flexible layer is transmissive to a selected wavelength of light.
7 . The article of claim 2 , wherein the bottom wall is substantially transmissive to the selected wavelength of light.
8 . The article of claim 6 , wherein the at least one sidewall is substantially nontransmissive to the selected wavelength of light.
9 . The article of claim 8 , wherein the at least one sidewall is at least 50% less transmissive of a selected wavelength of light than the bottom wall.
10 . The article of claim 8 , wherein a sidewall is at least 90% less transmissive of a selected wavelength of light than the bottom wall.
11 . The article of claim 1 , wherein the bottom wall and/or the at least one sidewall of a microwell further comprise a coating.
12 . The article of claim 11 , wherein the bottom wall and/or the at least one side wall of a microwell further comprises a plurality of coatings.
13 . The article of claim 11 , wherein at least one coating comprises SiO 2 .
14 . The article of claim 11 , wherein at least one coating comprises a reflective coating.
15 . The article of claim 1 , wherein the microstructured layer is a cured polymer derived from a resin.
16 . The article of claim 15 , wherein the resin is selected from the group consisting of acrylic-based resins derived from epoxies, polyesters, polyethers, and urethanes; ethylenically unsaturated compounds; aminoplast derivatives having at least one pendant acrylate group; polyurethanes (polyureas) derived from an isocyanate and a polyol (or polyamine); isocyanate derivatives having at least one pendant acrylate group; epoxy resins other than acrylated epoxies; and mixtures and combinations thereof.
17 . The article of claim 16 , wherein the optically transmissive layer comprises polyethylene terephthalate, polyethylene naphthalate, high density polyethylene, low density polyethylene, or linear low density polyethylene.
18 . The article of claim 16 , wherein the film comprises a multi-layer film.
19 . The article of claim 1 , wherein the optically-transmissive flexible layer further comprises an adhesive.
20 . The article of claim 1 , wherein the microstructured layer further comprises a tab region that is substantially free of microwells.
21 . The article of claim 20 , wherein the tab region comprises a detachable portion.
22 . The article of claim 1 , further comprising a cover layer coupled to the upper major surface of the microstructured layer.
23 . The article of claim 22 , wherein the cover layer is removably coupled to the microstructured layer.
24 . The article of claim 1 , further comprising a cover layer coupled to the optically-transmissive flexible layer on a surface opposite the microstructured layer.
25 . The article of claim 24 , wherein the cover layer is removably coupled to the optically-transmissive flexible layer.
26 . The article of claim 1 , further comprising an optical detection system comprising an optical device, wherein the optical device is optically coupled to a substrate.
27 . The article of claim 26 , wherein the optical device comprises a fiber optic face plate.
28 . The article of claim 26 , wherein the optical device comprises a CCD image sensor, a CMOS image sensor, or a photomultiplier tube.
29 . The article of claim 26 , wherein the optical system further comprises a processor.
30 . The article of claim 1 , wherein at least one microwell further comprises a polynucleotide.
31 . The article of claim 30 , wherein the polynucleotide is coupled to a particle.
32 . A process of manufacturing a microwell array article, comprising the steps of:
providing
a tool having a molding surface with a plurality of projections extending therefrom suitable for forming the microstructure elements;
a flowable, curable resin composition;
an optically-transmissive flexible layer having first and second major surfaces;
applying to the molding surface a volume of the flowable curable resin composition suitable for forming desired microstructure elements;
contacting the resin composition with the first major surface of the optically-transmissive flexible layer;
wherein the first major surface of the optically-transmissive flexible layer is surface-treated to promote adhesion to a cured resin composition;
wherein the thickness of the optically-transmissive flexible layer is about 50 μm or less; and
curing the resin composition while in contact with the flexible layer to form a microwell array article comprising a cured microstructured layer including a plurality of microwells bonded to the optically-transmissive flexible layer, and
removing the microwell array article from the tool.
33 . The process of claim 32 , wherein contacting the resin composition with the first major surface of the optically-transmissive flexible layer comprises applying pressure to the resin composition to substantially displace the resin between the tops of the projections in the tool and the surface of the optically-transmissive layer.
34 . The process of claim 32 , wherein the optically-transmissive flexible layer is surface-treated with a process selected from the group consisting of radiation treatment, corona discharge treatment, flame treatment, plasma treatment, high energy UV treatment, and chemical priming treatment.
35 . The process of claim 32 , wherein the optically-transmissive flexible layer is coupled to a carrier.
36 . The process of claim 32 , wherein the resin composition comprises a colorant.
37 . The process of claim 34 , wherein said curing comprises exposing the resin composition to at least one curing treatment selected from the group consisting of actinic radiation from a radiation source, an electron beam, and a chemical curing agent.
38 . The process of claim 32 , wherein the flexible layer has a thickness of about 2 μm to about 48 μm.
39 . The process of claim 32 , further comprising the step of removing a portion of the microstructured layer.
40 . The process of claim 32 , further comprising the step of removing a portion of the optically-transmissive flexible layer.
41 . The process of claim 32 , further comprising the step of disposing a reagent in a microwell.
42 . A method of detecting an analyte in a microwell array, comprising:
providing
a sample suspected of containing an analyte;
a reagent for the optical detection of the analyte;
an optical detection system; and
an article according to claim 1 ;
contacting the sample and the reagent in at least one microwells under conditions suitable to detect the analyte, if present, in the at least one microwells; and
using the optical detection system to detect the presence or absence of the analyte in a microwell.
43 . The method of claim 42 , wherein the optical system is optically coupled to the substrate.
44 . The method of claim 42 , wherein the optical system comprises a fiber optic face plate and wherein using the optical detection system comprises passing a signal through the fiber optic face plate.
45 . The method of claim 42 , wherein the optical system comprises a CCD image sensor, a CMOS image sensor, or a photomultiplier tube.
46 . The method of claim 42 , wherein the optical system further comprises a processor.
47 . The method of claim 42 , wherein detecting the presence or absence of an analyte comprises detecting light that is indicative of the presence of the analyte.
48 . The method of claim 47 , wherein detecting light comprises detecting light by absorbance, reflectance, or fluorescence.
49 . The method of claim 48 , wherein detecting light comprises detecting light from a lumigenic reaction.
50 . The method of claim 42 , wherein detecting the presence or absence of the analyte comprises obtaining an image of a microwell.
51 . The method of claim 50 , wherein detecting the presence or absence of the analyte comprises displaying, analyzing, or printing the image of a microwell.
52 . The method of claim 42 , wherein contacting the sample and the reagent in a plurality of microwells under conditions suitable to detect the analyte comprises an enzyme and an enzyme substrate.
53 . The method of claim 42 , wherein contacting the sample and the reagent in a plurality of microwells under conditions suitable to detect the analyte comprises forming a hybrid between two polynucleotides.
54 . An assay system, comprising:
an article according to claim 1 ;
an imaging device optically coupled thereto; and
a processor.
55 . The assay system of claim 54 , wherein optically coupled comprises adhesively coupling the microwell array article to a solid interface.
56 . A composition, comprising a compound selected from the group consisting of 1-(3-methyl-n-butylamino)-9,10-anthracenedione; 1-(3-methyl-2-butylamino)-9,10-anthracenedione; 1-(2-heptylamino)-9,10-anthracenedione; 1,1,3,3-tetramethylbutyl-9,10-anthracenedione; 1,10-decamethylene-bis-(-1-amino-9,10-anthracenedione); 1,1-dimethylethylamino-9,10-anthracenedione; and 1-(n-butoxypropylamino)-9,10-anthracenedione.
57 . The composition of claim 56 , further comprising a cured polymer.
58 . The composition of claim 57 , where the cured polymer is derived from a resin selected from the group consisting of acrylate resins, acrylic resins, acrylic-based resins derived from epoxies, polyesters, polyethers, and urethanes; ethylenically unsaturated compounds; aminoplast derivatives having at least one pendant acrylate group; polyurethanes (polyureas) derived from an isocyanate and a polyol (or polyamine); isocyanate derivatives having at least one pendant acrylate group; epoxy resins other than acrylated epoxies; and mixtures and combinations thereof.