# US 8,993,235 B2 · Encapsulated reagents and methods of use

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- Number: US 8,993,235 B2
- Document ID: US8993235B2
- Status: Granted
- Kind: B2
- Application number: 14/336,500
- Filed: 2014-07-21
- Published: 2014-11-13
- Granted: 2015-03-31
- Earliest claimed: 2009-01-16
- Applicants: Lee H. Angros
- Inventors: Lee H. Angros
- Web page: https://patentgrep.com/patent/US8993235B2

## CPC

- G01N 1/30: Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis}
- B01L 3/52: {Containers specially adapted for storing or dispensing a reagent (B01L3/02 takes precedence; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002)}
- B01L 3/523: {with means for closing or opening}
- C12Q 1/68: involving nucleic acids
- C12Q 1/6841: In situ hybridisation
- B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid
- B01L 2300/18: Means for temperature control
- B01L 2300/1861: using radiation
- G01N 35/1002: {Reagent dispensers}
- G01N 2035/00237: {Handling microquantities of analyte, e.g. microvalves, capillary networks}
- Y10S 435/962: Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
- Y10S 435/969: Multiple layering of reactants
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

The present invention contemplates use of encapsulated aqueous and non-aqueous reagents, solutions and solvents and their use in laboratory procedures. These encapsulated aqueous or non-aqueous reagents, solutions and solvents can be completely contained or encapsulated in microcapsules or nanocapsules that can be added to an aqueous or non-aqueous carrier solution or liquid required for medical and research laboratory testing of biological or non-biological specimens.

## Claims

### Claim 1

A method of treating a biological specimen on an analytic substrate, comprising:
disposing a liquid mixture upon an analytic substrate having a biological specimen disposed thereon and a residual wash buffer with at least one detergent present in the buffer disposed over the biological specimen, the liquid mixture comprising an oil-based carrier solution having dispersed therein a quantity of at least one of a plurality of microcapsules and a plurality of nanocapsules encapsulating at least one of a DNA probe and a RNA probe disposed in an aqueous solution containing at least one detergent; and
disrupting the at least one of a plurality of microcapsules and a plurality of nanocapsules to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution such that at least one of the DNA probe and the RNA probe and the aqueous solution separate from the oil-based carrier solution so as to become available to act on or react with the biological specimen and the residual wash buffer.

### Claim 2 (depends on 1)

The method of claim 1, wherein the disrupting step comprises exposing the at least one of a plurality of microcapsules and a plurality of nanocapsules to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.

### Claim 3 (depends on 1)

The method of claim 1, wherein the disrupting step comprises exposing the at least one of a plurality of microcapsules and a plurality of nanocapsules to a pressure differential sufficient to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.

### Claim 4 (depends on 1)

The method of claim 1, wherein the disrupting step comprises mechanically disrupting the at least one of a plurality of microcapsules and a plurality of nanocapsules to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.

### Claim 5 (depends on 1)

The method of claim 1, wherein the disrupting step comprises exposing the at least one of a plurality of microcapsules and a plurality of nanocapsules to a light energy to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.

### Claim 6 (depends on 1)

The method of claim 1, wherein the disrupting step comprises exposing the at least one of a plurality of microcapsules and a plurality of nanocapsules to electromagnetic radiation to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.

### Claim 7 (depends on 1)

The method of claim 1, wherein the disrupting step comprises exposing the at least one of a plurality of microcapsules and a plurality of nanocapsules to a chemical reaction to cause the release of the at least one of DNA probe and RNA probe and the aqueous solution into the oil-based carrier solution.