Abstract:
A system for processing and analyzing a sample includes a belt with wells that proceeds through the system, a dispensing station that dispenses the sample and reagents into the wells of the belt, and a detection station that detects an analyte in the wells of the belt. The system further includes a wash and decontamination station for decontaminating the wells of the belt.

Description:
CROSS-REFERENCE TO RELATED APPLICATION(S) 
       [0001]    This application claims priority from U.S. Provisional Application No. 61/792,556, filed Mar. 15, 2013 for “ARRAY TAPE FORMATION AND ARRAY TAPE BELT” by Darren Lynn Cook et al. 
     
    
     BACKGROUND 
       [0002]    The present invention relates to inline sample processing on high throughput systems, and more specifically relates to formation of a tape with a matrix of wells and a reusable belt with a matrix of wells. 
         [0003]    Advances in the biosciences industry have created a demand for high throughput biological sample processing and detection systems. For example, Astle, U.S. Pat. No. 6,632,653, discloses a high throughput method of performing biological assays using a tape with a matrix of wells. In a high throughput system, a liquid handling and sample processing system transfers the source and assay from microplates into a tape with a matrix of wells, seals the tape, and accumulates the tape on spools. The tape containing samples, such as biological samples, is then transferred to a water bath product and a reaction may be performed, such as polymerase chain reaction (PCR) using thermocycling. Subsequently, the tape may be loaded onto a detection instrument, which detects presence of a desired analyte, such as nucleic acid presence in a biological sample. 
         [0004]    Tape with a matrix of wells employed in such high throughput systems is typically used once to process and detect the presence of an analyte in a single sample. After a single use, the tape is discarded. It is not reused due to contamination risks. Additionally, tape with a matrix of wells is typically formed through thermal embossing. Therefore, consumable materials like tape with a matrix of wells increase costs associated with high throughput systems due to the cost of the tape and waste disposal. With a push towards increasing reaction speeds to process even more samples at an even faster rate, tape costs could become prohibitively expensive. 
       SUMMARY 
       [0005]    A system for processing and analyzing a sample includes a belt with wells that proceeds through the system, a dispensing station that dispenses the sample and reagents into the wells of the belt, and a detection station that detects an analyte in the wells of the belt. The system further includes a wash and decontamination station for decontaminating the wells of the belt. 
         [0006]    A method for processing and analyzing a sample in a system includes advancing a belt wells through the system to a dispensing station, dispensing a sample and reagents into the wells of the belt, advancing the belt to a detection station of the system, and detecting an analyte in the sample in the wells of the belt. The method further includes advancing the belt to a wash and decontamination station of the system and decontaminating the wells of the belt. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0007]      FIG. 1A  is a top view of a matrix of wells formed on a substrate using a die cut technique. 
           [0008]      FIG. 1B  is a cross-sectional view of a tape with a matrix of wells formed on a substrate using a die cut technique, along line  1 B- 1 B in  FIG. 1A . 
           [0009]      FIG. 2A  is a top view of a tape with a matrix of wells formed on a substrate using a laser technique. 
           [0010]      FIG. 2B  is a cross-sectional view of a tape with a matrix of wells formed on a substrate using a laser technique, along line  2 B- 2 B in  FIG. 2A . 
           [0011]      FIG. 3A  is a top view of a tape with a matrix of wells formed on a substrate using a thick film depositing technique. 
           [0012]      FIG. 3B  is a cross-sectional view of a tape with a matrix of wells formed on a substrate using a thick film depositing technique, along line  3 B- 3 B in  FIG. 3A . 
           [0013]      FIG. 3C  is a cross-sectional view of a tape with a matrix of wells formed on a substrate using a thick film depositing technique. 
           [0014]      FIG. 4A  is a top view of a tape with a matrix wells formed on a substrate. 
           [0015]      FIG. 4B  is a cross-sectional view of a tape with a matrix wells formed on a substrate using an additive technique, along line  4 B- 4 B in  FIG. 4A . 
           [0016]      FIG. 4C  is a cross-sectional view of a tape with a matrix wells formed on a substrate using a subtractive etching technique. 
           [0017]      FIG. 5  is a schematic diagram of a high throughput system employing a reusable belt with a matrix of wells. 
           [0018]      FIG. 6A  is a bottom view of an embodiment of a reusable belt with a matrix of wells on the bottom of the belt. 
           [0019]      FIG. 6B  is a cross-sectional view of an embodiment of a reusable belt with a matrix of wells on the bottom of the belt, along line  6 B- 6 B in  FIG. 6A . 
           [0020]      FIG. 7A  is a top view of an embodiment of a reusable belt with a matrix of wells on the top of the belt. 
           [0021]      FIG. 7B  is a cross-sectional view of an embodiment of a reusable belt with a matrix of wells on the top of the belt, along line  7 B- 7 B in  FIG. 7A . 
           [0022]      FIG. 8A  is a top view of an embodiment of a reusable belt with a matrix of wells on the top of the belt. 
           [0023]      FIG. 8B  is a cross-sectional view of an embodiment of a reusable belt with a matrix of wells with wells on the top of the belt, along line  8 B- 8 B in  FIG. 8A . 
           [0024]      FIG. 9  is a schematic diagram of another embodiment of a high throughput system employing a reusable belt with a matrix of wells. 
           [0025]      FIG. 10  is a schematic diagram of another embodiment of a high throughput system employing a reusable belt with a matrix of wells. 
           [0026]      FIG. 11  is a schematic diagram of another embodiment of a high throughput system employing a reusable belt with a matrix of wells. 
           [0027]      FIG. 12  is a schematic diagram of another embodiment of a high throughput system employing a reusable belt with a matrix of wells. 
           [0028]      FIG. 13  is a schematic diagram of another embodiment of a high throughput system employing a reusable belt with a matrix of wells. 
       
    
    
     DETAILED DESCRIPTION 
       [0029]    A system disclosed herein, in one aspect, provides more cost-effective methods of forming disposable tape with a matrix of wells. This provides accurate and controllable methods to introduce wells, recesses, or channels into a substrate to form a tape with a matrix of wells. Another embodiment replaces disposable tape with a matrix of wells with a reusable belt with a matrix of wells. The reusable belt progresses through a high throughput system for biological sample processing and detection, but is not discarded once detection is complete. Instead, the belt progresses through a decontamination regimen in order to remove processed biological material to allow the belt to be re-used for biological sample processing and detection. The high throughput system performs inline sampling, where a biological material is dispensed, reagents are added, the samples may be incubated for a specified amount of time to carry out a reaction, and the reaction may be scanned by a detector to determine the amount of an analyte in the biological material. 
         [0030]      FIGS. 1A and 1B  are a top view and a cross-sectional view of tape  20  including matrix of wells  21 , substrate  22 , and die-cut top layer  24 . Matrix of wells  21  is formed on substrate  22  using a die cut technique. Substrate  22  may be a substrate suitable for use as a tape with a matrix of wells. Using die-cut technology, a patterned film is laminated to substrate  22 . In an alternative embodiment, hole-punching technology is used to laminate a patterned film to substrate  22 . The attachment of a patterned film to substrate  22  can form matrix of wells  21 , resulting in die-cut top layer  24  with a bottom layer of substrate  22 . Matrix of wells  20  may be formed in an array pattern on substrate  22 . In alternative embodiments, the attachment of a pattern to substrate  22  can form a matrix of recesses, channels, or chambers. In one embodiment, this results in a disposable tape with matrix of wells  21  that can be employed in a high throughput system. The disposable tape with matrix of wells  21  can hold or control fluids or materials deposited into matrix of wells  21  for use, for example, in a bioassay or chemical reaction. A cover layer may be applied to contain fluid or material deposits within matrix of wells  20 . 
         [0031]      FIGS. 2A and 2B  are a top view and a cross-sectional view of tape  25  including matrix of wells  26  and substrate  28 . Matrix of wells  26  is formed on substrate  28  using a laser technique. Substrate  28  may be a substrate suitable for use as a tape with a matrix of wells. Using laser technology, such as an excimer laser, matrix of wells  26  is lasered into substrate  28 . In an alternative embodiment, a carbon dioxide (CO2) laser is used. An excimer laser can controllably and accurately create matrix of wells  26  in substrate  28 , allowing for extremely accurate, simple or complex, geometries to be machined into a flat tape format. Matrix of wells  26  may be formed in an array pattern on substrate  28 . The size and volume of matrix of wells  26  can be controlled by the amount of material removed by the excimer laser. Additionally, well density can be controlled through use of the laser technique. In alternative embodiments, a matrix of recesses, channels, or chambers can be lasered into substrate  28 . In one embodiment, this results in a disposable tape with matrix of wells  26  that can be employed in a high throughput system. The disposable tape with matrix of wells  26  can hold or control fluids or materials deposited into the wells for use, for example, in a bioassay or chemical reaction. A cover layer may be applied to contain fluid or material deposits within matrix of wells  26 . 
         [0032]      FIGS. 3A-3C  are a side view and cross-sectional views of tape  29  including matrix of wells  30 , substrate  32 , and thick film top layer  34 . Matrix of wells  30  is formed on substrate  32  using a thick film depositing technique. Substrate  32  may be a substrate suitable for use as a tape with a matrix of wells. Thick film technology may be used to deposit a thick film layer in a pattern on substrate  32  to form matrix of wells  30  on substrate  32 , resulting in thick film top layer  34  with a bottom layer of substrate  32 . The pattern may be an array pattern. The bottom of matrix of wells  30  consists of exposed substrate  32 . In alternative embodiments, an array of recesses, channels, or chambers can be formed on substrate  32  by depositing a thick film layer in a desired pattern. In one embodiment, this results in a disposable tape with matrix of wells  30  that can be employed in a high throughput system. The disposable tape with matrix of wells  30  can hold or control fluids or materials deposited into matrix of wells  30  for use, for example, in a bioassay or chemical reaction. Cover seal  36  may be applied to contain fluid or material deposits within matrix of wells  30 . 
         [0033]      FIGS. 4A-4B  are a top view and a cross-sectional view of tape  37  including matrix of wells  38 , photoresist top layer  40 , and substrate  42 . Matrix of wells  38  is formed on substrate  42  using an additive technique. Substrate  42  may be a substrate suitable for use as a tape with a matrix of wells. Matrix of wells  38  may be formed in an array pattern on substrate  42 . A photoresist layer is added to substrate  42  in a pattern to form matrix of wells  38  on substrate  42 , resulting in photoresist top layer  40  with a bottom layer of substrate  42 . The bottom of matrix of wells  38  consists of exposed substrate  42 . In an alternative embodiment shown in a cross-sectional view in  FIG. 4C , tape  43  includes matrix of wells  44  and substrate  46 . Matrix of wells  44  may be etched into substrate  46  using a subtractive technique such as photochemical etching, plasma etching, vapor etching, particle etching, or any other suitable etching technique. Matrix of wells  44  may be etched in an array on substrate  46 . 
         [0034]    Etching may controllably and accurately create wells in substrates, allowing for extremely accurate, simple or complex, geometries to be machined into a flat tape format. The size and volume of the wells can be controlled by the amount of material removed by etching. Additionally, well density may be controlled. In alternative embodiments, an array of recesses, channels, or chambers can be formed. In one embodiment, this results in tape  37  with matrix of wells  38  or tape  43  matrix of wells  44  that may be employed in a high throughput system. Tape  37  or tape  43  may hold or control fluids or materials deposited into the wells for use, for example, in a bioassay or chemical reaction. A cover seal may be applied to contain fluid or material deposits within matrix of wells  38  or matrix of wells  44 . 
         [0035]    In an alternative embodiment to the techniques described in  FIGS. 1-4 , a mechanical embossing technique may be used to emboss dimples into a substrate and displace the substrate material into the surrounding area. This mechanical embossing could be performed using a stamping technique or any other suitable mechanical technique. Mechanical embossing may produce arrays of wells with a range of volumes. For example, small dimples may be produced for reactions encased in oil that require very low sample volumes. 
         [0036]    In an alternative embodiment, instead of forming wells, a secondary substrate or coating could be applied to a base substrate using any appropriate technique such that the substrate or coating is capable of capturing samples and reagents. The substrate or coating may, for example, be paper, fabric, or a gel such as a hydrogel or agarose gel. Samples may be added in specific locations on the substrate. The substrate could be preloaded with reagents for a desired chemical reaction and the sample can subsequently be added to the substrate. Alternatively, the substrate may be preloaded with a sample and reagents subsequently added to the substrate. 
         [0037]    All of the above techniques can produce a tape with a matrix of wells with a flat bottom. The substrate may be plastic, metal, ceramic, glass, or any other suitable substrate for the appropriate technique. If a cover seal is applied, the tape with a matrix of wells will include both a flat top and a flat bottom. This is advantageous over traditional tape with a matrix of wells, which does not have a flat bottom due to thermoformed wells. A flat top and flat bottom allow imaging techniques for detecting a desired analyte, for example, to be used via either the cover seal or through the bottom layer of the tape with a matrix of wells or both. In some embodiments, magnets, heaters, coolers, vibrations, or other interactive systems may be applied directly to the flat surfaces of either or both the cover seal and the bottom layer of the tape with a matrix of wells to manipulate the fluids or materials deposited in the tape with a matrix of wells wells. In other embodiments, either or both the cover seal and the bottom layer of the tape with a matrix of wells may be imparted with a coating or finish that is metallic, dielectric, refractive, reflective, or absorbent. 
         [0038]    The well formation techniques can accurately produce wells that accommodate very low volumes of fluids and particles, such as wells for microfluidic applications. The techniques minimally affect the material chemical properties of substrate materials. Furthermore, these techniques are fast, low cost, and allow flexibility in manufacturing. These techniques may be used to create different formats for the tape with a matrix of wells such as individual arrays, continuous carrier tape including arrays used in a reel-to-reel process, microplate arrays, or slide arrays. The techniques described may also be used to create a matrix of wells on the second aspect of the system disclosed herein, which is a reusable belt with a matrix of wells. 
         [0039]      FIG. 5  is a schematic diagram of high throughput system  50  employing reusable belt  52 . Reusable belt  52  is a continuous loop/belt of tape with a built-in matrix of reaction wells. Reusable belt  52  can be utilized to process a sample, such as a biological sample, in a high throughput system and subsequently be decontaminated and reused to process a new sample. High throughput system  50  includes reusable belt  52 , dispensing station  54 , detection station  56 , and wash/decontamination station  58 . Wash/decontamination station  58  may include wash step  60 , vacuum step  62 , decontamination step  64 , and drying step  66 . Wash/decontamination station  58  eliminates the need for disposing of tape due to contamination risks, and allows reusable belt  52  to be used for processing numerous biological samples. Wash/decontamination station  58  provides a tightly controlled environment to control waste products such as amplicons from contaminating other parts of the high throughput system and subsequent samples. In one embodiment, wash/decontamination station  58  may be contained within a separate, sealed chamber. In an alternative embodiment, negative pressure could be used within a separate, sealed chamber to inhibit the possible escape of biological material from wash/decontamination station  58 . 
         [0040]    A biological sample may be loaded into the matrix of wells of reusable belt  52  and any necessary reagents for a desired reaction may be loaded into the matrix of wells of the reusable belt  52  at dispensing station  54 . A reaction can subsequently take place and reusable belt  52  may proceed to detection station  56  where a desired analyte can be detected. Once the detection is complete, reusable belt  52  proceeds to wash/decontamination station  58 . The first step in wash/decontamination station  58  is to remove the completed reaction by washing out the matrix of wells of reusable belt  52  in wash step  60  and/or applying a vacuum in vacuum step  62 . In an alternative embodiment an air knife or a water knife may be used. In other embodiments, any combination of a wash, a vacuum, an air knife, and a water knife may be used. The reaction wells are systematically washed depending on how the biological sample is processed within high throughput system  50 , including DNA amplification, RNA amplification, protein detection, and small molecule detection. 
         [0041]    After the bulk reaction is removed, reusable belt  52  moves to more refined decontamination step  64 , where decontamination and/or sterilization is performed to ensure that DNA/RNA/protein products are completely removed from the reaction wells of reusable belt  52 . Biological products may be removed using chemical solutions like bleach, acid, or any other suitable chemical agent. In alternative embodiments, UV radiation, heat, or cold may be used to remove the biological products. In an alternative embodiment, a chlorine solution may be sprayed in or fogged in, or the reaction wells may be immersed in a chlorine solution. After decontamination step  64  is complete, reusable belt  52  proceeds to drying step  66  where the reaction wells of reusable belt  52  are dried to make sure there is no residual decontamination material in the wells that would inhibit reactions of new samples. Since reusable belt  52  is a continuous loop with reaction wells built in, while some wells are decontaminated in wash/decontamination station  58 , decontaminated wells could proceed through the rest of high throughput system  50  to simultaneously process another biological sample. 
         [0042]    As stated above, the reaction wells in reusable belt  52  may be formed by using any of the techniques referred to in relation to  FIGS. 1-4 . Reusable belt  52  may be made of stainless steel or any other suitable metal, which is resilient to rust and degradation. In an alternative embodiment, reusable belt  52  may be made of a material similar to disposable tape with a matrix of wells, such as a polymer. In other embodiments, reusable belt  52  may be made of any other suitable material that can operate in a flexible manner and cycle within high throughput system  50 . The format of the reaction wells of reusable belt  52  may be a traditional rectangular array, a radial array, a single well row, or any other matrix format suitable for processing a desired biological material sample. Additionally, reusable belt  52  may be continuous or may be segmented, similar to a bulldozer track. In alternative embodiments, reusable belt  52  may be made of discrete films that are somewhat connected. In other embodiments, reusable belt  52  may be made of array segments that are held together, for example, by a magnet, strapped together, or riveted together. 
         [0043]      FIGS. 6A and 6B  are a bottom view and a cross-sectional view of an embodiment of reusable belt  52  with matrix of wells  68  with matrix of wells  68  formed on the bottom of reusable belt  52 . Matrix of wells  68  may be formed in an array pattern on reusable belt  52 . Reusable belt  52  may be transparent such that each of the wells of matrix of wells  68  is visible from the top of reusable belt  52 . With matrix of wells  68  on the bottom of reusable belt  52 , a reagent can be dispensed from the bottom of reusable belt  52 , as shown in  FIG. 5 . Depending on the reaction taking place in the high throughput system, detection may occur from the bottom or the top of this embodiment of reusable belt  52 . 
         [0044]      FIGS. 7A and 7B  are a top view and a cross-sectional view of another embodiment of reusable belt  52  with matrix of wells  68  formed on the top of reusable belt  52 . Matrix of wells  68  may be formed in an array pattern on reusable belt  52 . The bottom of reusable belt  52  is flat, thus detection can occur from either the top of reusable belt  52  or the bottom of reusable belt  52 . 
         [0045]      FIGS. 8A and 8B  are a top view and a cross-sectional view of another embodiment of reusable belt  52 . Reusable belt  52  includes arrays  70  with matrix of wells  68  and belt portion  72 . Belt portion  72  of reusable belt  52  may be made of stainless steel containing gaps or windows. Arrays  70  are configured to be placed over the gaps or windows in reusable belt  52 . Arrays  70  may be made of a transparent material such that detection may occur from underneath reusable belt  52 . 
         [0046]      FIG. 9  is a schematic diagram of high throughput system  80  employing reusable belt  82 . High throughput system  80  includes top dispensing station  84 , detection station  86 , and wash/decontamination station  88 . High throughput system  80  also allows simultaneous reaction incubation and detection for processes such as real time polymerase chain reaction and isothermal polymerase chain reaction. Since reusable belt  82  is a continuous loop with reaction wells built in, while some wells are decontaminated in wash/decontamination station  88 , decontaminated wells could simultaneously proceed through the rest of high throughput system  80  to process another biological sample. 
         [0047]      FIG. 10  is a schematic diagram of high throughput system  90  employing reusable belt  92 . High throughput system  90  includes bottom dispensing station  94 , detection station  96 , and wash/decontamination station  98 . Wash/decontamination station  98  of this embodiment includes a wash basin to decontaminate the reaction wells of reusable belt  92 . 
         [0048]      FIG. 11  is a schematic diagram of high throughput system  100  employing reusable belt  102 . High throughput system  100  includes dispensing station  104 , detection station  106 , belt cleaning station  108 , cover seal belt  110 , and cover seal belt cleaning station  112 . A biological sample may be loaded into reusable belt  102  of high throughput system  100  and a reagent may be dispensed at dispensing station  104 . Subsequently, a cover seal may be placed from cover seal belt  110  onto reusable belt  102 , covering the reaction wells of reusable belt  102 . Reusable belt  102  then proceeds to detection station  106 , where detection of a desired analyte occurs. The cover seal is then removed by cover seal belt  110 , and cover seal belt  110  continues to cover seal belt clean station  112 . Reusable belt  102  proceeds to belt cleaning station  108 . Thus, both cover seal belt  110  and reusable belt  102  are cleaned and decontaminated and may be reused in high throughput  100  system to process additional biological samples. 
         [0049]      FIG. 12  is a schematic diagram of high throughput system  120  employing reusable belt  122 . High throughput system  120  includes two dispensing stations  124 , detection station  126 , belt cleaning station  128 , cover seal dispensing station  130 , cover seal removal station  132 , and incubation station  134 . High throughput system  120  does not use a reusable cover seal. Instead, a sample may be loaded or dispensed into reusable belt  122 , one or more reagents may be dispensed in one or more of dispensing stations  124 , a cover seal may be placed over reusable belt  122  by cover seal dispensing system  120 , and reusable belt  122  may proceed through high throughput system  120 . Incubation station  134  allows reusable belt  122  to accumulate, incubate at a constant temperature, or pass through to detection station  136  if incubation is unnecessary for the desired reaction to take place. 
         [0050]    After reusable belt  122  passes through detection station  126  and the biological sample analysis is complete, the cover seal is removed by cover seal removal station  132  and the used cover seal is taken up by a cover seal take up. The used cover seal is subsequently disposed. Reusable belt  122  proceeds to belt cleaning station  128 , where reusable belt  122  is washed and/or decontaminated and may subsequently be reused in high throughput system  120  to process additional biological samples. 
         [0051]      FIG. 13  is a schematic diagram of high throughput system  140  employing reusable belt  142 . High throughput system  140  includes three dispensing stations  144 , detection station  146 , belt cleaning station  148 , two cover seal dispensing stations  150 , two cover seal removal stations  152 , and incubation station  154 . A sample may be loaded or dispensed into reusable belt  142 , one or more reagents may be dispensed in one or more of the first two dispensing stations  144 , a cover seal may be placed over reusable belt  142  by the first cover seal dispensing station  150 , and reusable belt  142  may proceed through high throughput system  140 . Incubation station  154  reusable belt  142  to accumulate, incubate at a constant temperature, or pass through to the third dispensing station  144  or detection station  146  if incubation is unnecessary for the desired reaction to take place. 
         [0052]    The cover seal may subsequently be removed at the first cover seal removal station  150  and reusable belt  142  may proceed to the third dispensing station  144 , where additional reagents may be added to the reaction wells. The reaction may be resealed at the second cover seal dispensing station  150 , and reusable belt  142  may proceed to detection station  146 . After detection, the second cover seal may be removed, taken up, and subsequently disposed at the second cover seal removal station  152 . Reusable belt  142  proceeds to belt cleaning station  148 , where reusable belt  142  is washed and decontaminated and may subsequently be reused in high throughput system  140  to process additional biological samples. 
         [0053]    Although the present invention has been described with reference to preferred embodiments, workers skilled in the art will recognize that changes may be made in form and detail without departing from the spirit and scope of the invention.