Devices, Compositions and Methods Pertaining To Microscopic Analysis Of Microorganisms and Other Analytes of Interest

Abstract

This invention pertains to devices, compositions and methods that can be used for the rapid determination of microorganisms, cells and other analytes of interest (e.g. a nucleic acid target) as well as associated properties of said microorganisms, cells and analytes. For example, said devices, compositions and/or methods can be applied to the determination of a trait of a microorganism present in a sample. Said devices, compositions and methods utilize matrix-forming prolonged-dissolution hydrophilic polymer to encapsulate hybridization probes and optionally other assay reagents within two or more reagent zones and/or matrix zones disposed on the surface of a substrate. Each reagent zone and/or matrix zone can be designed as a separate assay. Thus, a plurality of assays can be performed on a single substrate. In some embodiments, devices can be supplied in a form ready for a customer to rapidly perform one or a plurality of assays.

1 - 36 . (canceled) 
     
     
         37 . An assay device comprising at least two component parts, wherein,
 1) a first of said at least two component parts comprises: i) a first substrate comprising a first surface; ii) at least two matrix zones disposed on said first surface of said first substrate wherein at least one of said matrix zones comprises: a) at least one peptide nucleic acid probe; and b) at least one matrix-forming prolonged-dissolution hydrophilic polymer; and   2) a second of said at least two component parts comprises a second substrate comprising a second surface.   
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . An assay device comprising at least two component parts, wherein,
 1) a first of said at least two component parts comprises: i) a first substrate comprising a first surface; ii) a matrix film disposed on said surface; iii) at least two reagent zones disposed within said matrix film, wherein, a) said matrix film is formed using at least one matrix-forming prolonged-dissolution hydrophilic polymer; and b) at least one reagent zone comprises at least one peptide nucleic acid probe; and   2) a second of said at least two component parts comprises: i) a second substrate comprising a second surface, and ii) a sample disposed on said second surface.   
     
     
         41 . (canceled) 
     
     
         42 . The assay device of  claim 37 , wherein said at least two matrix zones are in gel, semi-solid or solid form. 
     
     
         43 . The assay device of  claim 37 , wherein said first surface of said first substrate comprises from 2 to 20 matrix zones. 
     
     
         44 . The assay device of  claim 37 , further comprising a polymer cap disposed on the substrate and wherein one or more of the matrix zones is located between the polymer cap and the substrate. 
     
     
         45 - 47 . (canceled) 
     
     
         48 . The assay device of  claim 37 , wherein said matrix-forming prolonged-dissolution hydrophilic polymer is selected from the group consisting of: multi-subunit sugar copolymers, pullulan, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, carboxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose hydroxypropyl methyl cellulose, polyvinylalcohol(s), polyvinylpyrrolidones(s), polyacrylamide(s), polyacrylic acid(s), polyethyleneimines(s), pectin(s) or mixtures of any two or more of the forgoing. 
     
     
         49 - 51 . (canceled) 
     
     
         52 . The assay device of  claim 37 , wherein the first surface of the first substrate and the second surface of the second substrate are separated by a narrow gap. 
     
     
         53 . The assay device of  claim 37 , further comprising a liquid disposed between said first surface of said first substrate and said second surface of said second substrate wherein said liquid further comprises a sample. 
     
     
         54 . (canceled) 
     
     
         55 . The assay device of  claim 53 , wherein said liquid is water or an aqueous solution that may optionally comprise detergent and/or buffer. 
     
     
         56 - 59 . (canceled) 
     
     
         60 . A method comprising:
 A) providing an assay device comprising at least two component parts, wherein,
 1) a first of said at least two component parts comprises: i) a first substrate comprising a first surface; ii) at least two matrix zones disposed on said first surface of said first substrate wherein at least one of said at least two matrix zones comprises: a) at least one peptide nucleic acid probe; and b) at least one matrix-forming prolonged-dissolution hydrophilic polymer; and 
 2) a second of said at least two device component parts comprises: i) a second substrate comprising a second surface; and ii) a sample disposed on said second surface; and 
   B) disposing an aqueous liquid between said first surface of said first substrate and said second surface of said second substrate such that said first surface and said second surface are in liquid communication.   
     
     
         61 - 67 . (canceled) 
     
     
         68 . A method comprising:
 A) providing an assay device comprising at least two component parts, wherein,
 1) a first of said at least two component parts comprises: i) a first substrate comprising a first surface; ii) at least two matrix zones disposed on said first surface of said first substrate wherein each of said at least two matrix zones comprises: a) at least one peptide nucleic acid probe; and b) at least one matrix-forming prolonged-dissolution hydrophilic polymer; and 
 2) a second of said at least two device component parts comprises a second substrate comprising a second surface; and 
   B) disposing a sample and an aqueous liquid between said first surface of said first substrate and said second surface of said second substrate.   
     
     
         69 - 76 . (canceled) 
     
     
         77 . A method comprising:
 A) providing an assay device comprising at least two component parts, wherein,
 1) a first of said at least two component parts comprises: i) a first substrate comprising a first surface; ii) a matrix film disposed on said surface; iii) at least two reagent zones disposed within said matrix film, wherein, a) said matrix film is formed using at least one matrix-forming prolonged-dissolution hydrophilic polymer; and b) at least one reagent zone comprises at least one peptide nucleic acid probe; and 
 2) a second of said at least two device component parts comprises: i) a second substrate comprising a second surface; and ii) a sample disposed on said second surface; and 
   B) disposing an aqueous liquid between said first surface of said first substrate and said second surface of said second substrate such that said first surface and said second surface are in liquid communication.   
     
     
         78 - 88 . (canceled) 
     
     
         89 . The method of  claim 68 , wherein one or more of said reagent zones further comprises a polymer cap disposed on the substrate and wherein said one or more reagent zones is located between the polymer cap and the substrate. 
     
     
         90 . (canceled) 
     
     
         91 . The method of  claim 68 , wherein first surface of said first substrate comprises from 2 to 20 matrix zones. 
     
     
         92 . The method of  claim 68 , wherein one or more of said matrix zones further comprise a buffer. 
     
     
         93 - 96 . (canceled) 
     
     
         97 . The method of  claim 68 , wherein said matrix-forming prolonged-dissolution hydrophilic polymer is selected from the group consisting of: multi-subunit sugar copolymers, pullulan, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, carboxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose hydroxypropyl methyl cellulose, polyvinylalcohol(s), polyvinylpyrrolidones(s), polyacrylamide(s), polyacrylic acid(s), polyethyleneimines(s), pectin(s) or mixtures of any two or more of the forgoing. 
     
     
         98 - 100 . (canceled) 
     
     
         101 . The method of  claim 68 , wherein the first surface of the first substrate and the second surface of the second substrate are separated by a narrow gap. 
     
     
         102 . The method of  claim 68 , further comprising a liquid disposed between said first surface of said first substrate and said second surface of said second substrate, wherein said liquid comprises the sample. 
     
     
         103 - 105 . (canceled)