Analytical biochemistry system with robotically carried bioarray
An analytical biochemistry system featuring a substrate with reactants immobilized thereon at fixed, known locations, a holder supporting the substrate and a manipulator for transporting the holder to a fixed sample and to an inspection station. The reactants are binding agents for a target biomolecule in a sample which forms a bound substance having a detectable characteristic. The holder may be a standard pipettor, optionally carried by a robot arm or hand as the manipulator to contact the sample for detection of the presence of target biomolecules within the sample. In one embodiment, the holder is a pipette tip within which the substrate is housed, or it may be a pipette adapter which bears the substrate and fits within the sample wells of a standard microtiter plate.
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This invention relates to a system and methods for detecting the presence of target biomolecules within samples with robotic assistance for a sample holder carrying an array of reactants.
BACKGROUND ARTAssays for the detection of target biomolecules within a sample, especially of multiple target biomolecules within a sample, are often performed by applying a volume of the sample to a test slide, membrane, or other substrate having immobilized reactants which may interact with the target or targets to form detectable complexes. These immobilized reactants are usually disposed at fixed locations, with samples brought to these locations. U.S. Pat. No. 5,139,743, for example, discloses a biochemical analysis apparatus wherein an applicator takes up a liquid sample and applies the sample to a fixed position test film for chemical analysis of the sample.
Sometimes complexes of target biomolecules and reactants are visually detectable directly after an appropriate incubation period for the sample and reactants, or after numerous development steps wherein development chemicals, such as fluorescent dye-conjugated molecules, are allowed to interact with the complexes. For example, the detection mechanism in U.S. Pat. No. 5,296,194 involves optically detecting a color change in a blood drop applied to a test slide.
U.S. Pat. No. 4,877,745 discloses methods for preparing immobilized reagents and applying samples to immobilized reagents. In particular, this patent discloses dispensing precisely controlled volumes of droplets onto a medium at precisely controlled locations, to form arrays of immobilized reagents by a jet head. An x-y plotter may be modified to carry a jet head so that reagent may be dispensed over an area.
Robotic laboratory workstations, such as the Biomek 1000 and 2000 of Beckman Instruments, Inc. have been developed for automatically carrying out assays involving multiple reactants and multiple samples. Typically such workstations are designed to deliver robotically precise volumes of reactants to a number of different samples located at known areas within the workstation. Alternatively, workstations can robotically move samples to reagents.
U.S. Pat. No. 5,171,537 to Wainwright et al. teaches activated immunodiagnostic pipette tips. The pipette tip houses a spherical element which is coated with a single ligand having affinity for a target molecule of a sample. With this device, the test element may be brought to contact the sample, as by aspirating the sample into the pipette tip. These pipette tips are limited in their sample throughput because they house only a single ligand reagent and thus preclude the detection of multiple analytes within a sample.
A class of devices known as optical biosensors, characterized by immobilized assay species within a supporter and a light collection device coupled to an optical waveguide, is also known. Optical biosensors may be used for detecting and quantifying the presence of specific species in test fluid samples, such as in clinical diagnostic reactions. For example, U.S. Pat. No. 4,857,273 discloses an optical biosensor for immunoassays and certain other reactions. Other examples, involving use of an optical fiber, are U.S. Pat. No. 5,143,066 and U.S. Pat. No. 5,401,469.
It is an object of the present invention to provide apparatus and methods for rapidly and automatically determining the presence of multiple target biomolecules in a single sample. It is another object of the present invention to provide analytical methods which require minimal sample volume and a minimal number of liquid transfers. It is a further object of the present invention to provide a device and system for rapid assessment of samples for target biomolecules which is readily adaptable to a variety of chemical and other detection schemes.
DISCLOSURE OF THE INVENTIONThe present invention achieves the above objects by providing an analytical biochemistry system for automated analysis of samples for the presence of target biomolecules. The system includes a solid substrate which is supported by a holder and carried by a manipulator, such as a robotic arm. Immobilized on the solid substrate surface at discrete, site-specific locations are reactants in an array which are capable of binding with target biomolecules in specific binding reactions to form immobilized biomolecule complexes. Such an array is termed a “bioarray”. The presence of target biomolecules in the sample is determined by detecting immobilized biomolecule complexes on the bioarray with some kind of probe, e.g. a fluorescence detector. In operation, the manipulator moves the bioarray to contact the substrate surface with a volume of sample. Then the manipulator moves the contacted bioarray to a detection station to detect the absence or presence of immobilized biomolecule complexes. In alternative embodiments the bioarray is stationary and a sample manipulator moves samples to the holder.
In the preferred embodiment, the bioarray is mobile, being carried by a manipulator. A detection station is located near the sample to probe the substrate after interaction between the reactants and sample or samples has occurred.
Distinct reactants specific to different target biomolecules are immobilized on a preferably flat, non-porous substrate. These reactants form a plurality of active sites on the substrate at known locations. The substrate may be a planar strip with linearly-arranged reactants forming separable spots or bands, or may be a planar sheet having an area-wide arrangement of reactants, forming spots or dots in a two-dimensional array, or may be a fiber or rod with substrate disposed in a manner similar to a strip.
The holder supports the bioarray and is carried by the manipulator which transports the substrate to the location of the fixed sample, and then to the location of the detection assembly. As stated, the substrate could be fixed and the sample transported. One example of a holder is a pipette or a pipette tip, within which a bioarray is affixed. The sample is drawn up into the pipette tip, as with aspiration from a bulb or vacuum pump, or withdrawal of a plunger. The sample is thus placed in contact with the substrate, allowing any target molecules which may be present within the sample to interact with the appropriate reactive sites on the substrate. After the appropriate incubation or reaction period, the sample may be removed from the pipette tip, as by air pressure or positive displacement with a plunger.
Another example of a useful holder is a pipette adapter resembling a truncated pipette tip and having a bracket or a flat surface for supporting the substrate. The pipette adapter may be placed directly into a sample, such as in a well of a microtiter plate or in a vial, in order to provide contact of the holder and the sample. The pipette adapter and accompanying substrate are then removed from the sample to a detector station. The various holders of the present invention may be adaptations of standard pipetting tools. The holders also are designed to require minimal sample volumes and to allow optical inspection of the substrate with minimal interference by the holder.
The method for detecting target biomolecules within a sample includes the steps of treating a substrate with a plurality of distinct reactants to form reagents immobilized on the substrate at fixed, known positions defining an array, i.e. a bioarray. The reactants are selected to bind one or more target molecules to form a complex having a detectable and identifiable characteristic, such as a fluorescent signature. The bioarray is supported in the holder. In turn, the holder has a shape which can be picked up by a manipulator which moves the substrate for contact with the fixed sample, and then removes and possibly rinses the substrate at another location to remove unbound biomolecules. Then the manipulator moves the substrate to a probing station, such as an optical inspection location for probing the active sites of the substrate with a beam for determining complementation of the target biomolecules by detecting the optically detectable characteristic.
Inspection may include detection of fluorescence, light scattering, absorbance, reflectance, chemiluminescence, radioactive emission, conductivity or electronic property. Depending on the nature of the substrate, detection of transmitted light is also possible. Prior to probing, intermediary steps to enhance visualization or realization of complementation, such as treatment with development chemicals, fluorescent dyes, etc. may be desired. Optical inspection of the substrate within the pipette tip is possible by use of an optical surface on the pipette tip. Optical inspection on the pipette adapter is unencumbered.
A manipulator in the form of a robotic arm gripping the pipette tip or pipette adapter type of substrate holder may place the bioarray in contact with the sample, and subsequently transfer the substrate to a detection assembly. Multiple sample transfers are thus eliminated. A computer controlling the robotic arm movement, the incubation times, and providing further analysis or display of detected signals from the substrate is preferred. An automated instrument includes a detection assembly, which in one embodiment includes a laser source providing an excitation beam to impinge upon the active sites of the substrate, a light collector for gathering signals emitted from the substrate, and a detector, such as a photomultiplier tube or CCD array. Alternatively, it may have multiple detection assemblies, depending on the requirements of the sample and the substrate chemistries. Relative movement of an excitation beam and the bioarray may be provided by the robotic arm holding the substrate or by scanning optics, such as a galvo mirror, within the excitation path of the detection assembly.
A substrate intended for use in the present invention may be an oligonucleotide array, a peptide array, or an immunochemical array, among others, and may be created on a separate member, such as a small slide, and affixed to the holder, or it may be created directly on the holder. Creation of the bioarray may be via biopolymer synthesis on a solid phase member or deposition of reactants, e.g. by movable nozzles, such as the type used for ink jet printing, or by some other method. The reactants may be affixed to the member via specific or non-specific covalent linkages, physical adsorption, or some other form of adhesion. The interaction or complexing of the target biomolecules and the immobilized reactants may be by affinity linkages, ionic linkages, adsorption, or some other reasonably secure manner.
The present invention provides a simple, highly adaptable method and apparatus for quickly and easily assessing samples for the presence of biomolecules.
BRIEF DESCRIPTION OF THE DRAWINGS
With reference to
In
By way of example, the reactants forming the active sites may comprise complementary DNA strands for detection by DNA hybridization or they may comprise immunological biomolecules for detection by immunological complexing, such as formation of antigen-antibody complexes.
The device of
In
In
The bioarray is preferably oriented so that the active sites face downward. Thus, when the adapter is placed within a sample well, as in
In
In
The bioarray is optically probed by the beam for determining the extent of complexing of the reactants in the active sites of the substrate with target biomolecules in the sample. The optical inspection may be for fluorescent signals, reflectance, absorbance, light scattering, or chemiluminescence, among others. Details of the optical system may vary according to the nature of the signal to be detected.
Arrow A of
The substrate may be formed by the device shown in
To perform a synthesis, a solid support material, such as a sheet of activated polypropylene, may be placed on top of the channels of the block. A backing plate may be used to sandwich the polypropylene substrate, allowing the flexible polypropylene to seal against the channels 50 of the block 45. The backing plate 52 of
With reference to
In
Another method of preparing the bioarray is by a technique analogous to a printing method. In this technique an analyte is deposited on a substrate by stamping or embossing a very thin layer with an array of analyte spots at desired locations. For example, an antigen attached to a molecule anchored to the substrate by pressure contact will combine with an appropriate antibody associated with a specific target biomolecule. The antibody may be fluorescent for optical detection.
Other methods of preparing the substrate may be used, particularly photolithographic techniques. In a journal article entitled “Light-Directed, Spatially Addressable Parallel Chemical Synthesis” by S. Fodor et al. in Science, Feb. 15, 1991, p. 767, the authors describe a method of synthesizing complex compounds at spatially discrete locations defined by photomasks of the type used in the semiconductor industry. Molecular building blocks are deposited at desired locations by exposing underlying building blocks, i.e. “deprotecting” the underlying block for a reaction with the superposed building block. Successive building blocks are added until a desired compound is formed. The location of each compound is precisely known from the mask set and the sites may be very closely spaced, limited only by the diffraction of light.
A method of imaging, i.e. probing, a substrate having microscopic features is by means of condensation figures (CFs) described in a journal article entitled “Imaging of Features on Surfaces by Condensation Figures” by G. Lopez et al. in Science, Apr. 30, 1993, p. 647. The authors describe the formation of an array of tiny droplets on a cold surface having an array of spots which are not wet by the droplets. The spots could be the complex compounds described in the preceding paragraph. The droplets are observed with microscope optics.
Still another bioarray forming technique is described in an article by B. Healey et al. in Science, Aug. 25, 1995 p. 1078. The authors deposited microscopic polymer arrays on a flat substrate by depositing a layer of polymerization solution on a flat plate, such as a glass chip which had been activated for adhesion with the solution. A bundle of fibers was brought into contact with the solution and then backed off and the substrate rinsed. Light was directed into the non-contacted end of the fiber bundle to cause polymer deposition on the substrate below the fibers of the fiber bundle. Polymer spots of a 2.0 micrometer diameter and a spacing of 4.0 micrometers were produced.
Yet another bioarray forming technique is the Southern blotting method in which hybridization is used simultaneously on a large number of DNA segments. DNA is fragmented, electrophoresed, denatured and transferred from a gel to filter paper. Positions of numerous fragments are established. The DNA fragments are robotically moved in accord with the present invention and combined with radioactive phosphorous labelled RNA which can be identified. The degree of DNA-RNA complementation, i.e. probing of the sample, can be determined by autoradiography.
In another bioarray forming technique a polyunsaturated polymerized lipid layer is applied to a support. The lipids have a member of a specific binding pair bound to one end. The lipids have an optical characteristic which is modified upon complexing the other member of the binding pair. Such an optical characteristic can be polarization of light and such light is used to probe the bioarray.
In
Although the method of the present invention is designed for detection of target biomolecules in a sample, quantification of the target biomolecules is possible by, for example, recording the sample volume exposed to the substrate, quantifying the degree of complementation at the active sites of the substrate, and calculating the amount of target biomolecule present from these two values. Quantification of the degree of complementation may be performed, e.g., by measuring the percentage of active sites which are fluorescently-labeled or give some other optical signal indicating complementation. Additionally, affixing an excess amount of reactants to the substrate compared to the amount of suspected target biomolecules of the sample is a preferred practice and makes quantification more accurate.
Referring to
In operation, the instrument of
In the above description, the robotic arm moved the pipette adapter, with holder and bioarray, to a sample location, such as a microtiter plate. However, the robotic arm could pick up sample in a pipettor and bring it to a stationary holder where the pipettor could dispense the sample onto the holder. Then, the same robotic arm, or another one, with an appropriate gripper could move the holder to a detection station.
The detection station could be any of the optical types described above, but could also be a radioactive tag detector if the immobilized reactants for the target biomolecule had been radioactive. Also, if the tag was a moiety suitable for detection by laser desorption mass spectrometry (LD-MS), then an LD-MS detection system could be used. Other tags and detection systems will be evident to those skilled in the art.
Claims
1. A system for detecting the presence of a target biomolecule within a sample comprising:
- a support surface treated with distinct reactants immobilized thereon in a spaced-apart relation, forming a substrate having a plurality of spaced-apart active sites, at least one of the reactants being reactive with a target biomolecule to form a bound complex having a detectable characteristic,
- holder means for supporting the substrate,
- an inspection station having means for probing the spaced-apart active sites of the substrate for said detectable characteristic of the sample, and
- manipulator means for bringing the holder means into contact with the sample and into said inspection station.
2-76. (canceled)
77. An apparatus for conducting chemical reactions, said apparatus comprising: (a) a plurality of wells in a housing and (b) a channel in said housing, said channel surrounding said plurality of wells and adapted for being filled with an amount of a fluid to form a convex meniscus extending above the top of said channel.
78. An apparatus according to claim 77 wherein said plurality of wells is in the form of a pattern in said housing.
79. An apparatus according to claim 77 wherein each of said wells has variable depth.
80. An apparatus according to claim 79 wherein each of said wells has a fluid circulation mechanism associated therewith.
81. An apparatus according to claim 80 wherein said fluid circulation mechanism comprises a member selected from the group consisting of sources for generating a thermal gradient causing convective flow and sources of mechanical energy.
82. An apparatus according to claim 81 wherein said member is a source for generating a thermal gradient selected from the group consisting of electrical current sources, infrared radiation sources, radio frequency sources, electrical resistance heaters, and semiconductor junction heaters, and Peltier cooling devices or said member is a source of mechanical energy selected from the group consisting of electric pulses, audio mechanical pulses, sub-audio mechanical impulses, vibration sources, ultrasonic pulses, and continuous wave acoustic sources.
83. An apparatus according to claim 77 wherein the surface properties surrounding each of said wells is different than the surface properties of said wells.
84. An apparatus according to claim 77 wherein each of said wells has a groove around its perimeter.
85. A method for conducting chemical reactions, said method comprising: (a) placing one or more liquid samples in separate wells in a housing surface comprising a plurality of said wells wherein the volume of said liquid sample in each of said wells is sufficient to form a convex meniscus at the surface of each of said wells, and (b) contacting said liquid samples with a plurality of arrays of chemical compounds wherein each of said arrays corresponds to a respective well in said housing.
86. A method according to claim 85 wherein said liquid samples are contacted with a substrate surface having a plurality of arrays of chemical compounds arranged on said substrate surface wherein each of said arrays corresponds to a respective well in said housing and wherein said substrate surface compresses each convex meniscus without cross-contact between adjacent liquid samples.
87. A method according to claim 86 further comprising, during said contacting, forming a seal between said substrate surface and said housing surface around the perimeter of said wells.
88. A method according to claim 87 wherein said seal is selected from the group consisting of fluid seals and seals is formed by placing a liquid in a channel in said housing surface surrounding said wells prior to contacting said substrate surface with said housing surface wherein the amount of said liquid is sufficient to form a convex meniscus.
89. A method according to claim 86 wherein said plurality of wells is in the form of a pattern in said housing.
90. A method according to claim 86 further comprising circulating said liquid sample in each of said wells.
91. A method according to claim 90 wherein the bottom of said wells is slanted and said method further comprises a step generating a thermal gradient causing convective flow in said liquid samples or a step of applying mechanical energy to said liquid samples sufficient to cause circulation therein.
92. A method according to claim 91 wherein said step is generating a thermal gradient causing convective flow selected from the group of steps consisting of (i) applying heat to said liquid samples sufficient to cause circulation in said samples from a heat source selected from the group consisting of electrical current sources, infrared radiation sources, radio frequency sources, electrical resistance heaters, and semiconductor junction heaters, and (ii) cooling said samples by means of a Peltier cooling device sufficient to cause circulating in said liquid samples or said step is applying mechanical energy selected from the group of steps consisting of (i) applying an electrical pulse to said liquid samples sufficient to cause circulating in said liquid samples, (ii) applying an audio or sub-audio mechanical impulse or vibration to said liquid samples sufficient to cause circulating in said liquid samples, and (iii) applying an ultrasonic pulse or continuous wave acoustic signal to said liquid samples sufficient to cause circulating in said liquid samples.
93. A method according to claim 86 wherein the surface properties surrounding each of said wells is different than the surface properties of said wells.
94. A method according to claim 86 wherein each of said wells has a groove around its perimeter.
95. A method according to claim 86 wherein said chemical reactions involve biopolymers.
96. A method according to claim 86 further comprising reading the arrays.
97. A method according to claim 96 comprising forwarding data representing a result obtained from reading one of the arrays.
98. A method according to claim 97 wherein the data is transmitted to a remote location.
99. A method according to claim 97 comprising receiving data representing a result of an interrogation obtained by reading one of the arrays.
100. A method of testing multiple liquid samples with multiple biopolymer arrays, said method comprising: (a) placing each of a multiple liquid samples in all or less than all separate wells in a housing surface comprising a plurality of said wells wherein the volume of said liquid sample in each of said wells is sufficient to form a convex meniscus at the surface of each of said wells, wherein the bottom of said wells is slanted, (b) placing a liquid in a channel in said housing surface surrounding said wells wherein the amount of said liquid is sufficient to form a convex meniscus, (c) contacting said liquid samples with a substrate surface having multiple biopolymer arrays arranged on said substrate surface wherein each of said arrays corresponds to a respective well in said housing and wherein said substrate surface compresses each convex meniscus without cross-contact between adjacent liquid samples and wherein forming a seal between said substrate surface and said housing surface around the perimeter of said wells, (d) causing circulation in said liquid samples, and (e) observing said substrate surface for the presence of reactions between said biopolymer arrays and said liquid samples.
101. A method according to claim 100 wherein said plurality of wells is in the form of a pattern in said housing.
102. A method according to claim 100 wherein said circulation is caused by generating a thermal gradient causing convective flow in said liquid samples or by applying mechanical energy to said liquid samples sufficient to cause circulation therein.
103. A method according to claim 102 wherein said circulation is caused by generating a thermal gradient causing convective flow selected from the group of steps consisting of (i) applying heat to said liquid samples sufficient to cause circulation in said samples from a heat source selected from the group consisting of electrical current sources, infrared radiation sources, radio frequency sources, electrical resistance heaters, and semiconductor junction heaters, and (ii) cooling said samples by means of a Peltier cooling device sufficient to cause circulating in said liquid samples or said circulation is caused applying mechanical energy selected from the group of steps consisting of (i) applying an electrical pulse to said liquid samples sufficient to cause circulating in said liquid samples, (ii) applying an audio or sub-audio mechanical impulse or vibration to said liquid samples sufficient to cause circulating in said liquid samples, and (iii) applying an ultrasonic pulse or continuous wave acoustic signal to said liquid samples sufficient to cause circulating in said liquid samples.
104. A method according to claim 100 wherein the surface properties surrounding each of said wells is different than the surface properties of said wells.
105. A method according to claim 100 wherein each of said wells has a groove around its perimeter.
106. A method according to claim 100 wherein said biopolymers are polynucleotides or polypeptides.
107. A kit for analyzing multiple biopolymer arrays on the surface of a substrate, said kit comprising in packaged combination: (a) an apparatus for conducting chemical reactions, said apparatus comprising: (i) a plurality of wells in a housing and (ii) a channel in said housing, said channel surrounding said plurality of wells and adapted for being filled with an amount of a fluid to form a convex meniscus extending above the top of said channel, and (b) a substrate having on a surface thereof a plurality of biopolymer arrays.
108. A method for conducting chemical reactions, said method comprising: (a) placing one or more liquid samples in separate wells in a housing surface comprising a plurality of said wells wherein each of the wells has a depth which varies within the well, and (b) contacting said liquid samples with a plurality of arrays of chemical compounds wherein each of said arrays corresponds to a respective well in said housing.
109. A method according to claim 108 wherein said liquid samples are contacted with a substrate surface placed over well openings, and which surface has the plurality of arrays of chemical compounds arranged on said substrate surface.
110. An integrated microfluidic array device comprising: a microfluidic component having a microfluidic feature for carrying a fluid of interest; and an array component comprising a flexible array substrate supporting an addressable collection of probes, said array component in operable association with said microfluidic feature of the microfluidic component for analyzing said fluid of interest.
111. The integrated microfluidic array device of claim 110 wherein said flexible any substrate comprises a flexible base supporting, in order, a reflective layer, a transparent layer, and the addressable collection of probes.
112. The integrated microfluidic array device of claim 110 wherein said flexible array substrate further comprises a flexible support supporting the flexible base.
113. The integrated microfluidic array device of claim 110 wherein the microfluidic component includes an identifier.
114. The integrated microfluidic array device of claim 110 wherein the integrated microfluidic array device is flexible.
115. The integrated microfluidic array device of claim 110 wherein the integrated microfluidic array device is rigid.
116. The integrated microfluidic array device of claim 110, wherein said microfluidic component further comprises a reservoir for holding one of a washing buffer, a sample fluid, or a reagent solution, the reservoir in fluid communication with the microfluidic feature.
117. The integrated microfluidic array device of claim 110, wherein said microfluidic component further comprises a valve for controlling fluid flow, the valve in operable relation to the microfluidic feature.
118. The integrated microfluidic array device of claim 110 wherein said flexible array substrate comprises a polymer material selected from the group consisting of polyacrylamide, polyetheretherketone, polyacrylate, polymethacrylate, polyesters, polyolefins, polyethylene, polytetrafluoroethylene, polypropylene, poly (4-methylbutene), polystyrene, poly(ethylene terephthalate), nitrocellulose, cellulose acetate, poly (vinyl chloride), polyamides, nylon, poly(vinyl butyrate), cross-linked dextran, and agarose.
119. The integrated microfluidic array device of claim 110 wherein said microfluidic component and said array component are modular.
120. The integrated microfluidic array device of claim 110 wherein said microfluidic feature is a microfluidic sipper structure.
121. The integrated microfluidic array device of claim 110, wherein said microfluidic feature is adapted to perform at least one of a reaction, concentration, or separation process.
122. The integrated microfluidic device of claim 110 wherein the microfluidic component includes a chamber in fluid communication with the microfluidic feature, the flexible array substrate disposed within the chamber.
123. The integrated microfluidic array device of claim 110 further comprising a chamber defined in part by the flexible array substrate, the chamber in fluid communication with the microfluidic feature.
124. The integrated microfluidic array device of claim 110 wherein the array component comprises a plurality of addressable collections of probes, each addressable collection of probes associated with its own separate chamber defined in pan by the array substrate.
125. The integrated microfluidic array device of claim 110 wherein more than one array component is joined to the microfluidic component.
126. The integrated microfluidic array device of claim 110 wherein more than one microfluidic component is joined to the array component.
127. The integrated microfluidic array device of claim 110, the array component further comprising an interface surface and the microfluidic component further comprising a mating surface, wherein the interface surface and the mating surface are adapted to fit together to form a fluid tight seal when the army component is in operable association with the microfluidic component.
128. The integrated microfluidic device of claim 127 wherein the array component is bonded to the microfluidic component using an adhesive.
129. The integrated microfluidic device of claim 127 wherein the array component is joined to the microfluidic component using ultrasonic welding.
130. The integrated microfluidic array device of claim 110, wherein said microfluidic component is adapted for interfacing to a standard multi-well plate.
131. The integrated microfluidic array device of claim 110 further comprising fiducial marks supported on the flexible array substrate.
132. The integrated microfluidic array device of claim 110 wherein the flexible array substrate has a thickness of less than about 800 microns.
133. A method of producing an integrated microfluidic array device comprising: a) forming a plurality of arrays by fabricating a plurality of addressable collections of probes on a flexible array substrate; b) forming a microfluidic component having a microfluidic feature; and c) placing at least one of said arrays in fluid communication with said microfluidic feature of said microfluidic component.
134. The method of claim 133, further comprising, before step c), cutting the flexible array substrate to provide said at least one of said arrays for step c).
135. A method of performing a hybridization assay on a sample comprising: a) obtaining an integrated microfluidic array device comprising a microfluidic component having a microfluidic feature, the microfluidic array device further comprising an array comprising a flexible array substrate supporting an addressable collection of probes in fluid communication with said microfluidic feature; b) introducing said sample to the integrated microfluidic array device; and c) contacting the addressable collection of probes with the sample for a time and under conditions sufficient to allow the sample to react with the probes, thereby performing the hybridization assay.
136. The method of claim 135, fisher comprising interrogating the array.
137. The method of claim 135, further comprising removing the array from the integrated microfluidic device prior to interrogating.
138. The method of claim 135, wherein the microfluidic array device further comprises a cover covering the array, the method further comprising removing the cover prior to interrogating the array.
139. The method of claim 135 further comprising mixing said sample during said reaction.
140. The method of claim 135 further comprising washing said array.
141. The method of claim 135 further comprising reading an identifier on the microfluidic device.
Type: Application
Filed: Mar 4, 2005
Publication Date: Mar 16, 2006
Applicant: Affymetrix, Inc. (Santa Clara, CA)
Inventors: Peter Coassin (San Juan Capristrano, CA), Jack McNeal (Long Beach, CA), David Helphrey (Santa Ana, CA)
Application Number: 11/073,288
International Classification: G01N 33/53 (20060101); B01L 3/02 (20060101);