# US 2014/0154664 A1 · Microfluidic apparatus and method of enriching target material in biological sample by using the same

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- Number: US 2014/0154664 A1
- Document ID: US20140154664A1
- Status: Published application
- Kind: A1
- Application number: 13/941,117
- Filed: 2013-07-12
- Published: 2014-06-05
- Earliest claimed: 2012-12-04
- Assignees: Samsung Electronics Co., Ltd.
- Applicants: Samsung Electronics Co., Ltd.
- Inventors: Min Seok S. KIM; Jong-myeon PARK
- Web page: https://patentgrep.com/patent/US20140154664A1

## CPC

- C12N 5/0693: {Tumour cells; Cancer cells}
- B01L 3/502753: {characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation}
- C12N 15/1006: {by means of a solid support carrier, e.g. particles, polymers}
- B01L 2200/0621: Control of the sequence of chambers filled or emptied
- B01L 2200/16: Reagents, handling or storing thereof
- B01L 2300/0803: Disc shape
- B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
- B01L 2400/0409: centrifugal forces
- B01L 2400/0677: phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
- B01L 2400/086: using baffles or other fixed flow obstructions
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

A micro fluidic apparatus includes a sample chamber that divides a sample into a first fluid layer and a second fluid layer due to a centrifugal force; a first enriching unit a that receives the first fluid layer from the sample chamber, forms a first complex of a first fine particle and the first target material, and separates the first complex from the first fluid layer using a density difference; and a second enriching unit that receives the second fluid layer from the sample chamber, forms a second complex of a second fine particle and the second target material, and separates the second complex from the second fluid layer using a density difference.

## Claims

### Claim 1

A microfluidic apparatus mounted on a rotation driving unit, the microfluidic apparatus comprising:
a sample chamber that receives a biological sample and separates the biological sample into a first fluid layer comprising a first target material and a second fluid layer comprising a second target material using centrifugal force;
a first enriching unit that receives the first fluid layer from he sample chamber, forms a first complex of a first fine particle and the first target material, and separates the first complex from the first fluid layer using a density difference with a density-gradient material (DGM) having a lower density than the first complex; and
a second enriching unit that receives the second fluid layer from the sample chamber, forms a second complex of a second fine particle and the second target material, and separates the second complex from the second fluid layer using a density difference between the second fluid layer and the second complex.

### Claim 2 (depends on 1)

The microfluidic apparatus of claim 1, wherein the first enriching unit comprises:
a first particle chamber that houses the first fine particle and is connected to the sample chamber by a first sample channel to receive the first fluid layer from the sample chamber;
a separation chamber that houses the DGM and is connected to the first particle chamber by a separation channel to receive the first fluid layer including the first complex; and
a first recovering chamber that is connected to the separation chamber by a first recovering channel to receive the first complex separated from the first fluid layer.

### Claim 3 (depends on 2)

The microfluidic apparatus of claim 2, wherein the first enriching unit further comprises a first waste chamber that is connected to the separation chamber by a first discharge channel to receive a fluid above the first complex in the separation chamber.

### Claim 4 (depends on 2)

The microfluidic apparatus of claim 2, wherein the first enriching unit further comprises:
a first cleaning solution chamber that houses a first cleaning solution and is connected to the separation chamber by a first cleaning channel to supply the first cleaning solution to the separation chamber; and
a first waste chamber that is connected to the separation chamber by a plurality of connection channels to receive a fluid above the first complex in the separation chamber.

### Claim 5 (depends on 2)

The microfluidic apparatus of claim 2, wherein the second enriching unit comprises:
a second particle chamber that houses the second fine particle and is connected to the sample chamber by a second sample channel to receive the second fluid layer from the sample chamber; and
a second recovering chamber that is connected to the second particle chamber by a second recovering channel to receive the second complex separated from the second fluid layer.

### Claim 6 (depends on 5)

The microfluidic apparatus of claim 5, wherein the second enriching unit further comprises a second waste chamber that is connected to the second chamber by a second discharge channel to receive a fluid above the second complex in the second particle chamber.

### Claim 7 (depends on 5)

The microfluidic apparatus of claim 5, wherein the second enriching unit further comprises:
a second cleaning solution chamber that houses a second cleaning solution and is connected to the second particle chamber by a second cleaning channel to supply the second cleaning solution to the second particle chamber; and
the second waste chamber that is connected to the second particle chamber by a plurality of connection channels to receive a fluid above the second complex in the second particle chamber.

### Claim 8 (depends on 1)

The microfluidic apparatus of claim 1, wherein the sample is blood, the first fluid layer is a white blood cell layer, and the second fluid layer is a plasma layer.

### Claim 9 (depends on 1)

The microfluidic apparatus of claim 1, wherein the first target material is a circulating tumor cell, a cancer stem cell, or a cancer cell.

### Claim 10

The microfluidic apparatus of claim wherein the second target material is nano (or micro)-vesicles, miRNA, DNA, or sub-micron sized lipid particles.

### Claim 11 (depends on 1)

The microfluidic apparatus of claim 1, wherein a partition wall is provided in the sample chamber to form a bottleneck portion together with at least one of a top wall and a bottom wall of the sample chamber to partially restrict the flow of the biological sample in a radial direction of the sample chamber, and
a gap between the bottleneck portion and the at least one of the top wall and the bottom wall is greater than a gap that would induce a capillary phenomenon.

### Claim 12 (depends on 11)

The microfluidic apparatus of claim 11, wherein the partition wall has such a shape that the bottleneck portion is narrower at an inner portion of the sample chamber than at an outer portion of the sample chamber in the radial direction of the sample chamber.

### Claim 13 (depends on 11)

The microfluidic apparatus of claim 11, wherein a plurality of partition walls are arranged in a circumference direction of the sample chamber and the partition walls are spaced apart from each other with openings therebetween.

### Claim 14 (depends on 13)

The microfluidic apparatus of claim 13, wherein each of the openings is narrower at an inner portion of the sample chamber than at an outer portion of the sample chamber in the radial direction.

### Claim 15

A method of enriching a target material using a microfluidic apparatus, the method comprising:
forming a first fluid layer that comprises a first target material and a second fluid layer that comprises a second target material by centrifuging a sample in a sample chamber of the microfluidic apparatus;
transporting the first and second fluid layers to first and second enriching units, respectively;
mixing a first fine particle that specifically binds a surface marker of the first target material with the first fluid layer to form a first complex of the first target material and the first fine particle, and separating the first complex from the first fluid layer using a DGM with a density that is less than the density of the first complex but greater than the density of the first fluid layer;
mixing a second fine particle that specifically binds a surface marker of the second target material with the second fluid layer to form a second complex of the second target material and the second fine particle, and separating the second complex from the second fluid layer using a density difference between the second complex and the second fluid layer; and
transporting the first and second complexes to first and second recovering chambers, respectively.

### Claim 16 (depends on 15)

The method of claim 15 further comprising discharging an upper layer material of the first complex and an upper layer material of the second complex to first and second waste chambers, respectively, before transporting of the first and second complexes to the first and second recovering chambers.

### Claim 17 (depends on 16)

The method of claim 16 further comprising:
cleaning the first and second complexes by supplying first and second cleaning solutions to the first and second enriching units, respectively, after the discharging of the upper layer material of the first complex and the upper layer material of the second complex to the first and second waste chambers, respectively, and
discharging the first and second cleaning solutions to the first and second waste chambers,

### Claim 18 (depends on 15)

The method of claim 15, wherein the sample is blood, the first fluid layer is a white blood cell layer, and the second fluid layer is a plasma layer.

### Claim 19 (depends on 15)

The method of claim 15, wherein the first target material is a circulating tumor cell, a cancer stem cell, or a cancer cell.

### Claim 20 (depends on 15)

The method of claim 15, wherein the second target material is nano (or micro)-vesicles, miRNA, DNA, or sub-micron sized lipid particles.