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

An assay device comprising:

• i) a substrate comprising a surface;
• ii) at least two matrix zones disposed on said surface of said substrate wherein at least one of said at least two matrix zones comprises:
• a) at least one hybridization probe comprising a linked fluorescent label;
• b) at least one hybridization probe comprising a linked quencher moiety;
• and c) at least one matrix-forming prolonged-dissolution hydrophilic polymer, wherein each matrix zone is a gel, semi-solid or solid.

An assay device comprising:

• i) a substrate comprising a surface;
• ii) a film of matrix disposed on said surface of said substrate wherein said film of matrix comprises a matrix-forming prolonged-dissolution hydrophilic polymer and at least two reagent zones;
• and iii) one or more reagents disposed at each of said reagent zones, wherein said film of matrix is a gel, semi-solid or solid and at least one reagent zone comprises:
• a) at least one hybridization probe comprising a linked fluorescent label and at least one hybridization probe comprising a linked quencher moiety;
• or b) at least one peptide nucleic acid probe.

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 matrix zones comprises:
• a) at least one hybridization probe comprising a linked fluorescent label;
• b) at least one hybridization probe comprising a linked quencher moiety;
• and c) 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;
• B) disposing a sample and an aqueous liquid between said first surface of said first substrate and said second surface of said second substrate;
• C) waiting for a period of time sufficient for said matrix-forming prolonged-dissolution hydrophilic polymer to re-hydrate and for:
• a) said at least one hybridization probe comprising a linked fluorescent label;
• and b) said at least one hybridization probe comprising a linked quencher moiety present at said matrix zone to interact with each other and with said sample;
• and
• D) determining one or more conditions of interest of said sample based on how:
• a) said at least one hybridization probe comprising a linked fluorescent label;
• and b) said at least one hybridization probe comprising a linked quencher moiety interact with each other and with said sample at each matrix zone.

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 first 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 hybridization probe comprising a linked fluorescent label and at least one hybridization probe comprising a linked quencher moiety;
• 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;
• C) waiting for a period of time sufficient for said matrix-forming prolonged-dissolution hydrophilic polymer to re-hydrate and for;
• a) said at least one hybridization probe comprising a linked fluorescent label;
• and b) said at least one hybridization probe comprising a linked quencher moiety present at said reagent zone to interact with each other and with said sample;
• D) determining one or more conditions of interest of said sample based, at least in part, on how:
• a) said at least one hybridization probe comprising a linked fluorescent label;
• and b) said at least one hybridization probe comprising a linked quencher moiety interact with each other and with said sample at said reagent zone.

A kit comprising the assay device comprising:

• i) a substrate comprising a surface;
• ii) at least two matrix zones disposed on said surface of said substrate wherein at least one of said at least two matrix zones comprises:
• a) at least one hybridization probe comprising a linked fluorescent label;
• b) at least one hybridization probe comprising a linked quencher moiety;
• and c) at least one matrix-forming prolonged-dissolution hydrophilic polymer, wherein each matrix zone is a gel, semi-solid or solid.