# US 10,371,604 B2 · Biological sample preparation for testing

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- Number: US 10,371,604 B2
- Document ID: US10371604B2
- Status: Granted
- Kind: B2
- Application number: 15/233,801
- Filed: 2016-08-10
- Published: 2017-02-16
- Granted: 2019-08-06
- Earliest claimed: 2015-08-10
- Assignees: Life Technologies Corporation
- Applicants: Life Technologies Corporation
- Inventors: Mousumi Rath; Mark Shannon; Jason La
- Web page: https://patentgrep.com/patent/US10371604B2

## CPC

- G01N 1/04: in the solid state, e.g. by cutting
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- G01N 1/2813: {Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence)}
- G01N 1/286: {involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F)}
- B01L 3/50825: {Closing or opening means, corks, bungs}
- B01L 2300/042: Caps; Plugs
- B01L 2300/046: Function or devices integrated in the closure
- B01L 2300/0838: Capillaries
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 2001/028: {Sampling from a surface, swabbing, vaporising}
- G01N 2001/045: {Laser ablation; Microwave vaporisation}
- G01N 2001/284: {using local activation of adhesive, i.e. Laser Capture Microdissection}
- G01N 2001/2886: {Laser cutting, e.g. tissue catapult}
- 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)
- 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
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

In one embodiment, a method for processing a sample includes selecting a selected sample from a biological specimen, the selected sample being in contact with a first surface of a first substrate. The method also includes transferring the selected sample directly from the first surface to a container comprising an internal volume. The method also includes forming or providing a sample solution within the internal volume of the container by contacting the selected sample with a lysis mixture using a protocol. The method further includes performing an assay, experiment, or test on the sample solution while the sample solution disposed is within the internal volume of the container. In another embodiment, a method for processing a sample includes providing a selected sample comprising one or more cells. The method also includes transferring the selected sample into an internal volume of a container. The method also includes contacting the selected sample with a lysis mixture using a protocol to provide a sample solution, wherein the protocol comprises heating the sample solution to a first temperature that is greater than 37 degrees Celsius and less than or equal to 75 degrees Celsius.

## Claims

### Claim 1

A method for processing a sample containing one or more nucleic acids, comprising:
selecting a sample from a biological specimen or heterogeneous mixture of biological cells using an optical system, wherein the selected sample is in contact with a first surface of a first substrate and the biological specimen or heterogeneous mixture of biological cells is in contact with a second substrate comprising a second surface;
moving the first substrate away from the second surface such that the selected sample maintains contact with the first surface and the selected sample does not maintain contact with the second surface;
attaching the first substrate to a container such that the selected sample is disposed within an internal volume of the container; and
forming a sample solution within the internal volume of the container by contacting the selected sample with a lysis mixture using a protocol.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the optical system comprises is a microscope.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the optical system comprises a laser capture microdissection apparatus.

### Claim 4 (depends on 1)

The method according to claim 1, wherein transferring the selected sample comprises:
levitating the selected sample into the container; or
using gravity to move the selected sample from the first substrate to the container.

### Claim 5 (depends on 1)

The method according to claim 1, further comprising detaching the first substrate from the container.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the first substrate comprises one or more of a laser capture microdissection cap (LCM cap), a sample carrier, or an extraction device.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the container comprises a container cap and transferring the selected sample comprises transferring the selected sample from the first substrate to the container cap.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the container comprises a well or vial of a microtiter plate and the first substrate is attached to the well or vial to form the internal volume.

### Claim 9 (depends on 8)

The method of claim 8, further comprising a plurality of additional substrates that are geometrically similar or equivalent to the first substrate, wherein each of the substrates is attached to one of the wells of the microtiter plate.

### Claim 10 (depends on 8)

The method of claim 8, wherein the microtiter plate comprises 96 wells.

### Claim 11 (depends on 1)

The method according to claim 1, wherein the container is a first container, the method further comprising attaching the first substrate to the first container to provide a first closed container.

### Claim 12 (depends on 11)

The method according to claim 11, further comprising at least one of placing the first closed container on a holder and/or placing the first closed container in an incubator.

### Claim 13 (depends on 12)

The method of claim 12, further comprising a plurality of additional closed containers, each additional closed container comprising an additional substrate and an additional container that are geometrically similar or equivalent to the first substrate and the first container, wherein the first closed container and the additional closed containers are together placed on the holder and/or placed in the incubator.

### Claim 14 (depends on 1)

The method according to claim 1, wherein the protocol comprises heating the sample solution to a first temperature that is greater than 37 degrees Celsius and less than or equal to 75 degrees Celsius.

### Claim 15 (depends on 1)

The method according to claim 1, wherein the protocol comprises heating the sample solution to a first temperature that is greater than or equal to 45 degrees Celsius and less than or equal to 75 degrees Celsius.

### Claim 16 (depends on 1)

The method according to claim 1, further comprising performing an assay, experiment, or test on the sample containing one or more nucleic acids.

### Claim 17 (depends on 16)

The method according to claim 16, wherein the assay, experiment, or test on the sample containing one or more nucleic acids is performed without filtration, purification, or pipetting of the sample solution.

### Claim 18 (depends on 16)

The method according to claim 16, wherein performing an assay, experiment, or test on the sample containing one or more nucleic acids comprises sequencing the one or more nucleic acids.

### Claim 19 (depends on 16)

The method according to claim 16, wherein performing an assay, experiment, or test on the sample containing one or more nucleic acids includes performing a polymerase chain reaction (PCR) assay, experiment or test on one or more nucleic acids.