Microfabricated, flowthrough porous apparatus for discrete detection of binding reactions

Abstract

An improved microfabricated apparatus for conducting a multiplicity of individual and simultaneous binding reactions is described. The apparatus comprises a substrate on which are located discrete and isolated sites for binding reactions. The apparatus is characterized by discrete and isolated regions that extend through said substrate and terminate on a second surface thereof such that when a test sample is allowed to the substrate, it is capable of penetrating through each such region during the course of said binding reaction. The apparatus is especially useful for sequencing by hybridization of DNA molecules.

1 - 20 . (canceled)  
   
   
       21 . A device for binding a target molecule, comprising: 
 a substrate having oppositely facing first and second major surfaces;    a multiplicity of channels extending through said substrate from said first major surface to said second major surface;    a first binding reagent immobilized on the walls of a first group of said channels; and    a second binding reagent immobilized on the walls of a second group of said channels.    
   
   
       22 . The device of  claim 21 , wherein the channels within the first or second group are branched or interconnected.  
   
   
       23 . The device of  claim 21 , wherein said first and second binding reagents differ from one another.  
   
   
       24 . The device of  claim 21 , wherein said substrate is fabricated from glass or silicon.  
   
   
       25 . The device of  claim 21 , wherein said binding reagents are selected from the group consisting of DNA, proteins and ligands.  
   
   
       26 . The device of  claim 21 , further comprising a rigid support, wherein said rigid support is integral to said substrate, or is bonded to said substrate.  
   
   
       27 . A device for binding a target molecule, comprising: 
 a substrate having oppositely facing first and second major surfaces;    a multiplicity of discrete regions of channels extending through said substrate from said first major surface to said second major surface; and    at least a binding reagent immobilized on the walls of a said discrete regions of channels.    
   
   
       28 . The device of  claim 27 , wherein the channels within the group of said channels are branched or interconnected.  
   
   
       29 . The device of  claim 27 , wherein said substrate is fabricated from glass or silicon.  
   
   
       30 . The device of  claim 27 , wherein said binding reagents are selected from the group consisting of DNA, proteins and ligands.  
   
   
       31 . The device of  claim 27 , further comprising a rigid support, wherein said rigid support is integral to said substrate, or is bonded to said substrate.  
   
   
       32 . A method for detecting a binding reaction comprising: 
 a) providing a substrate having (i) oppositely facing first and second major surfaces, (ii) a multiplicity of channels extending through said substrate from said first major surface to said second major surface, and (iii) a binding reagent immobilized on the walls of a group of said channels;    b) passing a sample through the substrate; and    c) detecting binding between a binding target in the sample and the binding reagent on the, thereby detecting said binding reaction.    
   
   
       33 . The method of  claim 32 , wherein the channels within the first group are branched or interconnected.  
   
   
       34 . The method of  claim 32 , wherein said substrate is fabricated from glass or silicon.  
   
   
       35 . The method of  claim 32 , wherein said binding reagents are selected from the group consisting of DNA, proteins and ligands.  
   
   
       36 . The method of  claim 32 , wherein said binding reagents are oligonucleotide probes.  
   
   
       37 . The method of  claim 32 , wherein the substrate is contained in a sample well.  
   
   
       38 . The method of  claim 32 , wherein the detection step is accomplished by a CCD camera.  
   
   
       39 . A method for detecting binding reactions comprising: 
 a) providing a substrate having (i) oppositely facing first and second major surfaces, (ii) a multiplicity of discrete regions of channels extending through said substrate from said first major surface to said second major surface, (iii) a first binding reagent immobilized on the walls of a first group of said channels, and (iv) a second binding reagent immobilized on the walls of a second group of said channels;    b) passing a sample through the substrate; and    c) detecting binding of the fist and second binding reagents with first binding targets in the sample, thereby detecting said binding reactions.    
   
   
       40 . The method of  claim 39 , wherein the channels within the first or second group are branched or interconnected.  
   
   
       41 . The method of  claim 39 , wherein said substrate is fabricated from glass or silicon.  
   
   
       42 . The method of  claim 39 , wherein said binding reagents are selected from the group consisting of DNA, proteins and ligands.  
   
   
       43 . The method of  claim 39 , wherein the first group of channels is contained in a first sample well and the second group of channels is contained in a second sample well.  
   
   
       44 . The method of  claim 32 , wherein the detection step is accomplished by a CCD camera.