Substrates For Isolating, Reacting And Microscopically Analyzing Materials

Abstract

An immobilizing device for biological material comprises a rigid support ( 12 ) carrying a substrate layer ( 20, 20′ ) of polymer having biological immobilizing properties, e.g. for amino and nucleic acids. Substantially solid ultra-thin substrate layers ( 20′ ) having a thickness less than about 5 micron, preferably between about 0.1 and 0.5 micron, and micro-porous, ultra-thin substrate layers ( 20′ ) having a thickness less than about 5 micron, preferably less than 3 micron, 2 or 1 micron are shown, which may be segmented by isolating moats M. The substrate layer is on a microscope slide ( 302 ), round disc ( 122 ), bio-cassette, at the bottom of a well of a multiwell plate, and as a coating inside a tube.

1 . A device for immobilizing biological material comprising a polymer substrate layer  20 ,  20 ′,  20 ″ having biological immobilizing properties preferably for protein or nucleic acid, the substrate layer deposited on a rigid support  12 , and having an outer deposit-receiving region exposed to receive the biological material, wherein the substrate layer is ultra-thin, having a thickness tut less than about 5 micron. 
     
     
         2 . (canceled) 
     
     
         3 . The device of  claim 1  wherein the rigid support  12  defines a straight support surface e. g., a planar surface such as that of a microscope slide  10   a  or cylindrical surface, and said substrate layer  20 ,  20 ′,  20 ″ is a drawn coating applied directly or indirectly to the rigid support in the direction of said straight surface, preferably drawn substantially according to  FIG. 14 . 
     
     
         4 . The device of  claim 1  wherein the deposit-receiving region of said substrate layer  20 ,  20 ′, 20 ″ is in a surface-treated state for enhanced adhesion of deposits of biological material thereon, preferably the surface treatment being that provided by a corona treater  122 . 
     
     
         5 . The device of  claim 1  wherein at least one intervening layer  14 ,  14 ′,  112  lies between said rigid support  12  and said ultra-thin polymer substrate layer  20 ,  20 ′,  20 ″, said intervening layer adherently joined on each of its oppositely directed faces to substance of said device. 
     
     
         6 . The device of  claim 5 , wherein immediately adjacent materials on opposite sides of a said intervening layer  14 ,  14 ′,  112  are not as adhesively compatible with each other as each is with said intervening layer. 
     
     
         7 . The device of  claim 5 , wherein said intervening layer  14 ,  14 ′,  112  is an adherent oxide of metal, preferably an oxide of tantalum or aluminum, or a silica based material, e.g. colloidal silica or a soluble silicate such as sodium silicate. 
     
     
         8 . The device of  claim 5 , wherein a said intervening layer  14 ,  14 ′,  112  is of substance selected from the group consisting of silane, epoxy silane, polylisine, PEI, GAP, an adherent metal oxide, colloidal silica and soluble silicates. 
     
     
         9 . The device of  claim 5 , wherein a said rigid support  12  defines a straight support surface e.g., planar or cylindrical, and said intervening layer  14 ,  14 ′,  112  is a drawn coating applied directly or indirectly to the rigid member in the direction of said straight surface, preferably drawn substantially according to  FIG. 13 , preferably said intervening layer  14 ,  14 ′,  112  comprising colloidal silica or a soluble silicate. 
     
     
         10 . The device of  claim 1  wherein a surface of one of the constituents of said device, prior to being united with a next constituent layer of the device, is in a surface-treated state for enhanced adhesion of that surface to said next constituent layer. 
     
     
         11 . The device of  claim 5 , wherein a said intervening layer  14 , 14 ′, 112  is at least partially opaque, the intervening layer blocking at least 30%, preferably blocking at least 50%, or often preferably blocking at least 70% of incident radiation at a wave length corresponding to the stimulating or emission wavelength of a fluorophore tag on biological material. 
     
     
         12 . The device of  claim 10 , wherein the rigid support  12  has characteristic fluorescence or luminescence in response to incident stimulating radiation, a said intervening layer  14 ,  14 ′,  112  being effective to at least substantially limit penetration of incident stimulating radiation from the substrate layer  20 , 20 ′, 20 ″ to the support or limit penetration of fluorescent or luminescent radiation from the support to the substrate layer, or both. 
     
     
         13 . The device of  claim 5  wherein a said intervening layer  14 ,  14 ′,  112  is electrically conductive. 
     
     
         14 . The device of  claim 13  wherein an electric terminal is associated with said intermediate layer  14 ,  14 ′,  112  for applying a voltage potential to or sensing the potential of said layer. 
     
     
         15 . The device of  claim 14  constructed and arranged such that electrical potential applied to said terminal is effective to provide a potential to the substrate layer  20 ,  20 ′,  20 ″ of the device, to promote binding or rejection of biological material exposed to said substrate layer. 
     
     
         16 . The device of  claim 5  constructed and arranged to support biological material for microscopy, wherein a said intervening layer  14 ,  14 ′,  112  is an oxide of metal, preferably oxide of tantalum or aluminum, adapted to serve at least one of the functions of adhesively uniting, either directly or indirectly, said rigid support  12  with said deposit-receiving substrate layer  20 ,  20 ′,  20 ″, of providing an opaque barrier to prevent or substantially limit light passing between said deposit-receiving substrate layer  20 ,  20 ′,  20 ″ and said support  12 , or of providing an electrically conductive layer as a means e.g. to electrically charge said substrate layer  20 , 20 ′, 20 ″. 
     
     
         17 . The device of  claim 1  wherein the substrate layer  20 ,  20 ′,  20 ″ is adapted to receive a deposit of biological material  420  and to be temporarily engaged by an object adjacent the deposit, as by an elastomeric gasket  430 , wherein the substrate layer is interrupted so that adherence of the substrate to the removed object does not disrupt the array, preferably an intervening adhesion promoting layer  14 ′ beneath said substrate layer being interrupted such that a substrate layer  20 ″ applied thereto is disrupted, for example by moat M,  FIG. 23 ,  24 , formed by a gap in a pattern of a metal oxide adhesion promoting intervening layer  14 ′. 
     
     
         18 . The device of  claim 17  in which said substrate layer is applied as a continuous fluid coating which separates on drying at interruptions of an adhesion promoting layer, such as the metal oxide layer. 
     
     
         19 . The device of  claim 1  constructed for use in microscopy, wherein an outer surface of the substrate layer  20 ,  20 ′,  20 ″ is constructed to receive deposits of biological material thereon in position exposed for direct illumination and inspection from the exterior. 
     
     
         20 . The device of  claim 1 , wherein the device is functionally at least partially transparent to pass effective light in at least one direction between a deposit on said substrate layer  20 ,  20 ′,  20 ″ and through said rigid support  12 , for instance the device arranged to enable illumination of a deposit of biological material on said substrate layer  20 ,  20 ′,  20 ″ via said rigid support  12 , or the device arranged to enable microscopic inspection of a deposit of biological material on said substrate layer  20 ,  20 ′,  20 ″ via said rigid support  12 , or the device arranged to enable microscopic inspection of a deposit of biological material on said substrate layer from both the exterior side of said substrate layer side and via the rigid support. 
     
     
         21 . (canceled) 
     
     
         22 . A device for immobilizing biological material comprising a polymer substrate layer  20 , 20 ′, 20 ″ having biological immobilizing properties preferably for protein or nucleic acid, the substrate layer deposited on a rigid support  12 , and having an outer deposit-receiving region exposed to receive biological material, wherein at least one intervening layer  14 ,  14 ′,  112  lies between said rigid support and said polymer substrate layer, said intervening layer adherently joined to substance of said device on each of its oppositely directed faces and wherein a said intervening layer  14 ,  14 ′,  112  is at least partially opaque to radiation employed to stimulate emission from the biological material, limiting or preventing transmission of radiation from said rigid support, preferably wherein immediately adjacent materials on opposite sides of a said intervening layer  14 ,  14 ′,  112  are not as adhesively compatible with each other as each is with said intervening layer. 
     
     
         23 - 25 . (canceled) 
     
     
         26 . The device of  claim 1 , wherein said substrate layer  20 ′,  20 ″ is substantially solid, preferably having a thickness less than about 5 micron, preferably less than 3, 2 or 1 micron and in preferred embodiments between about 0.1 and 0.5 micron. 
     
     
         27 . The device of  claim 1 , wherein said substrate layer  20 , at least in its outer region, is micro-porous, preferably said substrate layer being microporous throughout its thickness, and preferably, said substrate layer having a thickness less than 3 micron, preferably less than 2 or 1 micron. 
     
     
         28 - 36 . (canceled) 
     
     
         37 . A method of forming the device of  claim 1  including applying directly or indirectly to said rigid support a fluid containing said polymer of said substrate layer under conditions to form said substrate layer, preferably by drawing the rigid support from a bath of coating composition. 
     
     
         38 . The method of  claim 37  including applying an adhesion-promoting layer directly or indirectly to said rigid support before application of said substrate layer, preferably by applying a metal, preferably tantalum or aluminum and allowing it to oxidize, or applying soluble silica or colloidal silicate by drawing from a bath. 
     
     
         39 - 48 . (canceled) 
     
     
         49 . A method of conducting an assay including providing a device according to  claim 1 , applying an array of spots of bio-material to the substrate, conducting an assay which tags at least some of the spots with a fluorescent label, and, after washing the array, reading the array by fluorescent detection, preferably the assay being based on protein-protein interaction, or involving an array comprising nucleic acid or other genetic material, or comprising viruses, peptides, antibodies, receptors, cDNA clones, DNA probes, oligonucleotides including synthetic oligonucleotides, or polymerase chain reaction (PCR) products, or the array comprising plant, animal, human, fungal or bacterial cells, or malignant cells, or cells from biopsy tissue. 
     
     
         50 . The method of  claim 49  wherein the reading is accomplished with a CCD sensor.