This invention relates to a chamber apparatus.
In order to conduct parallel expression profiling of hundreds to thousands of genes or proteins typically a microarray is utilized. A microarray is a collection of microscopic spots attached to a substrate in a defined pattern, with the substrate generally consisting of a slide, chip, or plate of glass, plastic, or silicon. The spots may be of DNA, biological or chemical samples, other nucleic acids, proteins, or other probe materials. The probes are immobilized in a predetermined pattern on the substrate, such that each probe has a defined position. Microarray-based assays typically include exposing the arrayed probes to fluidic samples that contain target materials, which may interact with specific probes on the microarray. In a nucleic acid microarray, for example, arrayed single-stranded synthetic oligonucleotide or cDNA probes are contacted with labeled (e.g., fluorescently, radioactively, etc.) single-stranded target nucleic acids, which hybridize with complementary probe molecules in the microarray. Since the probes are arrayed at predetermined positions, the presence and quantity of target sequences in the fluid can be identified by the position at which fluorescence or radiation is detected and the intensity of the emitted fluorescence or radiation, respectively.
Microarray technology provides a user with the ability to perform hundreds to thousands of parallel biological or chemical assays. This technology is applicable for basic and applied research. For basic research, microarray based assays are used in finding genes (e.g. by hybridizing cDNA to predict open reading frames) and in the identification of common regulatory elements (e.g. by gene co-expression), for example. In applied research, the technology is used, e.g., in complex system profiling (e.g., of specific organs and diseases, stress responses, aging, and wound healing) in disease diagnosis, prognosis, and classification, in performing toxicity assessments (e.g., of drugs, foods, environmental conditions, etc.), and in drug discovery (e.g., to identify and validate targets to optimize efficacy, etc.)
Microarrays are typically manufactured by synthesizing or dispensing probe material on the surface of a planer substrate. To conduct an assay, a fluid well is typically formed by addition of chamber on the top surface of the substrate. Currently, this configuration of microarray or multiple well plate assays includes an assembly containing the array itself, with a chamber to contain the target hybridization solution, and a separate gasket or adhesive to contain the solution in the wells and prevent leakage. This type of plate or assay configuration requires multiple components to be assembled, very accurate gasket placement, and sufficient pressure to hold and compress the gasket to the substrate without buckling. The requirement for utilization of multiple components makes it cumbersome to fully utilize the microarray or multiple well plates. In addition, there is a risk of incorrect gasket placement such that when a sample is loaded into an array or well plate, this sample may leak into another array or well plate, which prevents an accurate test from being conducted. Further, as assays are often conducted over extended time periods or at elevated temperatures, the chambers must be sealed to prevent evaporation.
Therefore, there are several features of a chamber design that are desirable. The interface between the substrate and chamber must form a seal to prevent fluid from leaking out. The chamber must be critically aligned with the probe features on the substrate. The chamber/substrate apparatus should be easy to assemble. Further, the chamber should be removable, to allow the substrate to be scanned or imaged in standard equipment. It is also beneficial for the chamber to be designed for ease of use during the assay, either by manual handing of an operator, or by integration with standard automation equipment. To this end, the formation of a top chamber surface that may be easily sealed, either manually or by an automated system, is required. There are currently no microarray chamber designs available that incorporate all of these features. The current invention describes a chamber apparatus with an integrated lower gasket to form a removable seal to the substrate, and a second integrated upper gasket to form a sealable surface to enclose the chamber with a chamber cover, thereby limiting evaporation.
The present invention has been accomplished in view of the above-mentioned technical background, and it is an object of the present invention to provide a chamber apparatus that may contain a single well or multiple wells that prevents inserted samples from leaking into other portions or other wells of the chamber apparatus. It is an additional object of the present invention to provide a chamber apparatus that is easily utilized and assembled, and also easily disassembled for further substrate processing.
In a preferred embodiment of the invention, a chamber apparatus is disclosed. A chamber frame has an integrated upper gasket and an integrated lower gasket, where a cover is disposed over the integrated upper gasket. The integrated upper gasket and the integrated lower gasket are disposed on the chamber frame by over-molding. A substrate is disposed below the chamber frame, where the substrate interfaces with the integrated lower gasket. A substrate frame is disposed below the substrate, where the substrate frame structure is configured to receive the substrate, where the substrate frame is aligned and fastened to the chamber frame that is configured to receive the substrate frame. A chamber cover is disposed onto the integrated upper gasket forming a leak tight seal and preventing evaporation of the assay fluid. The chamber cover may cover one or multiple chamber apparatuses, and may be placed or removed manually or by automation hardware.
In another preferred embodiment of the invention, a system for utilizing multiple chamber apparatuses is disclosed. A plurality of chamber apparatuses is inserted into chamber tray. The chamber tray is configured to receive the plurality of chamber apparatuses.
In yet another preferred embodiment of the invention, a method of assembling a chamber apparatus is disclosed. A chamber frame is provided. The chamber frame is disposed in between an integrated upper gasket and an integrated lower gasket by over-molding. A substrate frame and substrate are provided. A cover over the integrated upper gasket is provided. The substrate frame positions and contains the substrate. A chamber frame is placed over the substrate and substrate frame. The chamber frame is assembled into the substrate frame producing a leak tight seal between the substrate and individual well(s).
These and other advantages of the present invention will become more apparent as the following description is read in conjunction with the accompanying drawings, wherein:
The presently preferred embodiments of the invention are described with reference to the drawings, where like components are identified with the same numerals. The descriptions of the preferred embodiments are exemplary and are not intended to limit the scope of the invention.
The lower exterior portion of the frame 101 also includes a slide rail 101d that acts as a means to locate and for fixturing or fitting the chamber apparatus 100 with the chamber tray 609 for automation, etc as in
Chamber cover 101c is fabricated or produced by ordinary machining or machine injection molding processes. The chamber cover 101c is fabricated by using a molding or a machining process and it is disposed or assembled to 101 by snapping it in place manually. Critical variables such as draft and mold temperatures must be considered when utilizing this molding process. The cover or chamber cover 101c is utilized for hybridization to prevent evaporation by forming a compression seal with the upper integrated gasket. The lower integrated gasket, 101b, seal prevents samples, specimens or biomolecules (nucleic acids, proteins etc.) from leaking into other wells after the specimen is inserted into a well or chamber 101e (
Referring to
These barcode labels 101f and 103a can be read by a wide range of commercial optical scanners. As an alternative, the labels may include an RFID tag or transponder, which can be read by scanners that utilize radio frequency identification (RFID) technology.
In
In
Referring to
Next, at block 807 the seal is removed and the well 101e of the chamber apparatus 100 is flushed and another solution is added to well 101e. For example, the well 101e is flushed three times with 0.75×TNT and 250 ul of 0.75×TNT is added into the well 101e and the well 101e is sealed again. The chamber apparatus 100 is incubated at 46 degrees Celsius for 1 hour. At block 809, the seal is removed from the well 101e and the solution is removed from the well 101e. Next, 250 ul of staining solution is then added, then the solution is incubated at ambient temperature for 30 minutes in a dark area. At block 811, the array well 101e is flushed three times, then 1×TNT is added in each well 101e and incubated at an ambient temperature for twenty minutes in a dark area. Next the 1×TNT is removed and the wells filled with a low salt final rinse buffer. At block 813 the final rinse solution is removed and the substrate is dried, by placing a chamber tray with the chamber apparatus into a centrifuge bucket and spinning until dry. The chamber apparatus 100 is placed in a light-tight box until scanning. The chamber apparatus may be dissembled to allow removal of the substrate 103, which may then be scanned by a suitable scanning or imaging device.
This invention provides a system that allows a user to assemble a chamber apparatus that prohibits samples from leaking or mixing with other samples or chamber array wells, when they are inserted into an array well of the chamber apparatus. In addition, the chamber frame design allows for easy assembly and disassembly for simplified use and slide substrate scanning on conventional microarray scanners. Chamber apparatus includes a chamber frame with an upper integrated gasket and a lower integrated gasket, a substrate, and a substrate frame that positions and captures the substrate. The lower integrated gasket provides a single sealing surface between the chamber frame and the substrate. The upper integrated gasket interfaces with a chamber cover forming a compression seal that prevents sample loss due to evaporation during the hybridization process. The chamber frame and substrate frame have integrated features that allow them to align and fasten to each other by latching or snapping, resulting in an optimal clamping force to produce a compression seal between the integrated lower gasket and the substrate.
It is intended that the foregoing detailed description of the invention be regarded as illustrative rather than limiting and that it be understood that it is the following claims, including all equivalents, which are intended to define the scope of the invention.
This application claims priority to U.S. provisional patent application No. 60/806,108 filed Jun. 29, 2006; the disclosure of which is incorporated herein by reference in its entirety.
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/US07/72172 | 6/27/2007 | WO | 00 | 12/22/2008 |
Number | Date | Country | |
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60806108 | Jun 2006 | US |