Surface assembly for immobilizing DNA capture probes in genetic assays using enzymatic reactions to generate signal in optical bio-discs and methods relating thereto

Abstract

The invention relates to methods and systems for use of bio-discs in quantifying an amount or concentration of one or more substances that are present in a biological sample. The system includes an optical disc having one or more samples deposited thereon, an optical element configured to emit and divert radiation onto the samples and components for measuring spectral change of the compound and determining an amount or concentration of one or more target substances. The bio-disc is prepared using a method including providing a membrane that is or can be dimensional to fit within a channel of the bio-disc, applying a reagent on the membrane which is configured to allow the reagent to be released from the membrane into a solution in contact with the membrane, and depositing the membrane in a channel of the bio-disc.

1 . A method of preparing a bio-disc having at least one channel, the method comprising: 
 providing a membrane that is or can be dimensioned to fit within the at least one channel of the bio-disc;    applying one or more reagents on the membrane, wherein the membrane is configured to allow the one or more reagents to be released from the membrane into a solution placed in contact with the one or more reagents on the membrane; and    depositing the membrane in one of the at least one channels of the bio-disc.    
     
     
         2 . The method of  claim 1 , wherein the bio-disc comprises a semi-reflective layer having a thickness of less than about 400 Å.  
     
     
         3 . The method of  claim 1 , wherein the bio-disc comprises a semi-reflective layer having a thickness of between about 100 and 300 Å.  
     
     
         4 . The method of  claim 1 , wherein the bio-disc comprises a plurality of channels and a plurality of membranes are deposited in the plurality of channels.  
     
     
         5 . The method of  claim 1 , wherein the reagents on the membrane are allowed to dry before the depositing step is performed.  
     
     
         6 . The method of  claim 1 , wherein the one or more reagents comprise one or more enzymes.  
     
     
         7 . The method of  claim 1 , wherein providing a membrane comprises providing a membrane that is or can be dimensioned to fit within a side chamber of the bio-disc that is in fluid communication with the at least one channel.  
     
     
         8 . The method of  claim 1 , wherein the membrane is a bibulous hydrophilic material.  
     
     
         9 . The method of  claim 1 , wherein the membrane comprises hydrophilic polyethersulfone.  
     
     
         10 . The method of  claim 1 , wherein the applying is performed using a pipette.  
     
     
         11 . An apparatus for quantifying an optical density change in calorimetric assays, the apparatus comprising: 
 an optical disc having one or more compounds deposited thereon, wherein the one or more compounds change one or more spectral characteristics in the presence of a target substance so that a spectral change by each of the one or more compounds is a function of a concentration of the target substance brought into contact with each of the one or more compounds;    an optical element configured to emit and direct radiation so that the radiation is incident on the compounds;    a detector configured to measure an indication of the spectral change for each of the one or more compounds;    a computing device responsive to the detector and configured to determine an amount or concentration of one or more target substances.    
     
     
         12 . A method of quantifying an amount of one or more analytes present in a biological sample, the method comprising: 
 providing an optical disc having one or more reagents located on or in one or more analysis zones of the optical disc;    introducing a sample onto or into the optical disc so that the sample contacts the one or more reagents on the optical disc;    incubating the optical disc for a period of time;    quantifying a spectral change in at least one portion of the disc resulting from introduction of the sample; and    determining an amount of the one or more analytes present in the sample based upon results from the quantifying step.    
     
     
         13 . The method of  claim 12 , wherein the sample comprises one or more of a body fluid sample, an agricultural product sample, a food sample, a waste product sample, and an environmental sample.  
     
     
         14 . A method of quantifying an amount of one or more analytes present in a biological sample, the method comprising: 
 depositing one or more reagents onto respective one or more analysis zones on an optical disc;    applying a sample onto the optical disc so that the one or more reagents is brought into contact with the sample;    incubating the optical disc for a period of time;    emitting radiation having a known wavelength so that the radiation is incident upon the sample;    quantifying an amount of radiation transmitted through the sample in contact with each of the one or more reagents; and    determining an amount of the one or more analytes present in sample based upon results from the quantifying step.