# US 12,216,033 B2 · Contact-type patch, staining method using the same, and manufacturing method thereof

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- Number: US 12,216,033 B2
- Document ID: US12216033B2
- Status: Granted
- Kind: B2
- Application number: 18/351,796
- Filed: 2023-07-13
- Published: 2023-11-16
- Granted: 2025-02-04
- Earliest claimed: 2016-02-23
- Assignees: Noul Co., Ltd.
- Applicants: Noul Co, Ltd.
- Inventors: Dongyoung Lee; Kyunghwan Kim; Youngmin Shin; Hyunjeong Yang; Chanyang Lim
- Web page: https://patentgrep.com/patent/US12216033B2

## CPC

- G01N 1/312: {for samples mounted on planar substrates}
- B01F 33/3039: with mixing achieved by diffusion between layers
- B01L 3/00: Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- C07K 16/3061: {Blood cells}
- C12Q 1/6844: Nucleic acid amplification reactions
- C12Q 1/6848: characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 1/701: {Specific hybridization probes}
- G01N 1/2813: {Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence)}
- G01N 1/30: Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis}
- G01N 1/31: Apparatus therefor
- G01N 15/06: Investigating concentration of particle suspensions (by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10)
- G01N 15/14: Optical investigation techniques, e.g. flow cytometry
- G01N 21/77: by observing the effect on a chemical indicator
- G01N 21/78: producing a change of colour
- G01N 33/4833: {of solid biological material, e.g. tissue samples, cell cultures (tissue in vivo A61B5/00; cell suspensions G01N33/48735)}
- G01N 33/49: Blood {(chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80)}
- G01N 33/5082: {Supracellular entities, e.g. tissue, organisms}
- G01N 33/52: Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)}
- G01N 33/5304: {Reaction vessels, e.g. agglutination plates (for solid-phase systems G01N33/543)}
- G01N 33/533: with fluorescent label
- G01N 33/575: for cancer
- G01N 33/60: involving radioactive labelled substances
- G01N 35/00029: {provided with flat sample substrates, e.g. slides (G01N35/028 takes precedence)}
- G06T 7/0012: {Biomedical image inspection}
- G06T 7/0014: {using an image reference approach}
- B01L 3/505: {Flexible containers without fluid transport within}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- G01N 15/01: specially adapted for biological cells, e.g. blood cells (investigating sedimentation of particle suspensions in blood G01N15/05)
- G01N 15/075: by optical means
- G01N 2001/302: {Stain compositions}
- G01N 2021/7723: {Swelling part, also for adsorption sensor, i.e. without chemical reaction}
- G01N 2021/7786: {Fluorescence}
- G01N 2035/00039: {Transport arrangements specific to flat sample substrates, e.g. pusher blade}
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- Y02P 20/582: Recycling of unreacted starting or intermediate materials
- 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)
- B01F: MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING (mixing paints B44D3/06)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- 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
- G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present disclosure relates to a gel-phase patch that performs a function of assisting in staining during a staining process such as a process of coming into contact with a specimen such as blood to perform a staining function of staining the specimen, a process of fixing the specimen, or a process of forming an optimal pH at a specimen stained by a staining sample. According to an aspect of the present disclosure, a contact-type staining patch includes a staining solution that reacts with a specimen and a gel receptor provided as a gel matrix of a mesh structure in which a pore that accommodates the staining solution is formed and the mesh structure prevents the staining solution in the pore from leaking or degenerating, and having a contact surface that comes into contact with the specimen to transfer some of the staining solution to the specimen.

## Claims

### Claim 1

A method of staining a specimen using a gel-phase contact-type staining patch, the method comprising:
staining a specimen fixed on a slide with a staining reagent,
wherein, the staining step comprises,
contacting a surface of the gel-phase contact-type staining patch comprising a staining reagent configured to react with the specimen and a buffer solution to the specimen fixed on the slide; and
separating the gel-phase contact-type staining patch from the specimen.

### Claim 2 (depends on 1)

The method of claim 1, wherein the buffer solution has a predetermined pH value, wherein the predetermined pH value is identical to an optimal pH value of the staining reagent.

### Claim 3 (depends on 1)

The method of claim 1, wherein the buffer solution has a predetermined pH value, wherein the predetermined pH value is smaller than the optimal pH of the staining reagent when the optimal pH is acidic, and is larger than the optimal pH of the staining reagent when the optimal pH is basic.

### Claim 4 (depends on 1)

The method of claim 1, wherein the gel is any one selected from the group consisting of hydrogel, silicone gel, silica gel, agar gel, agarose gel, PDMS (polydimethylsiloxane) gel, and PMMA (polymethylmethacrylate) gel.

### Claim 5 (depends on 1)

The method of claim 1, wherein the contacting the surface of the gel-phase contact-type staining patch to the specimen fixed on the slide is performed by applying predetermined pressure.

### Claim 6 (depends on 1)

The method of claim 1, wherein the contacting the surface of the gel-phase contact-type staining patch to the specimen fixed on the slide is performed without applying any external pressure.

### Claim 7 (depends on 1)

The method of claim 1, wherein the staining reagent is any one selected from the group consisting of acetocarmine, methylene blue, eosin, Azure II, acidic fuchsin, crystal violet, safranin, Janus green B, hemotoxylin, Giemsa solution, Wright solution, Wright-Giemsa solution, Romanowski staining solution, Leishman staining solution, Gram staining solution, carbol-fuchsin, Ziehl solution, DNA probe coupled to DAPI (4,6-diamidino-2-phenylindole) fluorochorme, a fluorescent substance, an antibody coupled to an enzyme, an antibody coupled to a fluorescent substance, and an antibody coupled to an isotope.