# US 12,109,564 B2 · Micro-fluidic device for concentration of particles via centrifugal force, and corresponding centrifugation and/or detection device

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- Number: US 12,109,564 B2
- Document ID: US12109564B2
- Status: Granted
- Kind: B2
- Application number: 16/972,283
- Filed: 2019-06-05
- Published: 2021-08-05
- Granted: 2024-10-08
- Earliest claimed: 2018-06-06
- Assignees: ELTEK S.p.A.
- Applicants: ELTEK S.p.A.
- Inventors: Marco Pizzi; Giovanni Melioli; Valentina Gallo; Massimo Zanin
- Web page: https://patentgrep.com/patent/US12109564B2

## CPC

- B01L 3/502753: {characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation}
- G01N 1/4077: {by other techniques involving separation of suspended solids}
- C12Q 1/18: Testing for antimicrobial activity of a material
- G01N 35/00069: {whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk}
- B01L 2200/0668: Trapping microscopic beads
- B01L 2300/0645: Electrodes
- B01L 2300/0806: Standardised forms, e.g. compact disc [CD] format
- B01L 2300/0861: Configuration of multiple channels and/or chambers in a single devices
- B01L 2300/161: Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
- G01N 2001/4083: {sedimentation}
- G01N 2035/00495: {Centrifuges}
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- 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

## Abstract

A microfluidic device for concentrating particles contained in a fluid sample (FS) comprises a substrate (11) configured for being set in rotation about a centre of rotation (5a), the substrate (11) having a surface (11b) defined in which is at least one microfluidic arrangement (12) that extends substantially according to a plane identified by the substrate (11). The at least one microfluidic arrangement (12) comprises at least one microchannel (13) having a first end (13a) and a second end (13b), wherein: —the at least one microchannel (13) comprises, in a region thereof intermediate between its first end (13a) and its second end (13a), at least one accumulation area (15) that is at a first distance (R1) in a radial direction (R) from the centre of rotation (5a) of the substrate (11); —the first end (13a) and the second end (13b) of the at least one microchannel (13) are at second distances (R2) in a radial direction (R) from the centre of rotation (5a) of the substrate (11); and —the first distance (R1) in a radial direction (R) is greater than the second distances (R2) in a radial direction (R), in such a way that particles (P) possibly contained in a volume of fluid of the fluid sample (FS) that penetrates into the at least one microchannel (13) tend to concentrate in the at least one accumulation area (15) as a result of the centrifugal force caused by a rotation of the substrate (11) about the centre of rotation (5a).

## Claims

### Claim 1

A microfluidic device for concentrating and detecting particles contained in a fluid sample, the microfluidic device comprising:
a substrate configured for being set in rotation with respect to a centre of rotation, the substrate having a surface at which at least one microfluidic arrangement is defined, which extends substantially according to a plane identified by the substrate, wherein the at least one microfluidic arrangement comprises a plurality of microchannels each having a first end and a second end; wherein
each microchannel of the plurality of microchannels comprises, in a region thereof intermediate to the first end and the second end of the microchannel, at least one detection area that is at a first distance in a radial direction with respect to the centre of rotation of the substrate;
the first end and the second end of each microchannel of the plurality of microchannels are at second distances in the radial direction with respect to the centre of rotation of the substrate;
the first distance in the radial direction is greater than the second distances in the radial direction,
wherein the at least one detection area is an area for particle accumulation;
wherein the at least one microfluidic arrangement comprises a common loading duct or chamber and a common venting duct or chamber, all the microchannels of the plurality of microchannels being fluidically connected to the common loading duct or chamber and to the common venting duct or chamber, all the microchannels of the plurality of microchannels having the respective first ends and second ends fluidically connected in parallel to the common loading duct or chamber and to the common venting duct or chamber, respectively, the common loading duct or chamber being operable for receiving the fluid sample, such that the fluid sample can penetrate from the common loading duct or chamber into all the microchannels of the plurality of microchannels, and air contained in all the microchannels of the plurality of microchannels can progressively be vented at the common venting duct or chamber,
wherein, in the at least one detection area for particle accumulation, each microchannel of the plurality of microchannels is substantially V-shaped or U-shaped, or comprises two substantially convergent microchannel branches, the at least one detection area of each microchannel of the plurality of microchannels being free of exit ways for the fluid sample,
in such a way that particles contained in a volume of fluid of the fluid sample that penetrates into at least one microchannel of the plurality of microchannels concentrate in the at least one detection area as a result of the centrifugal force caused by a rotation of the substrate about the centre of rotation.

### Claim 2 (depends on 1)

The microfluidic device according to claim 1, wherein the microchannels of the plurality of microchannels are arranged substantially side by side in the radial direction.

### Claim 3 (depends on 1)

The microfluidic device according to claim 1, wherein each microchannel of the plurality of microchannels is shaped, at the respective at least one detection area, to define at least one of the following:
at least one syphon,
at least one well.

### Claim 4 (depends on 1)

The microfluidic device according to claim 1, wherein each microchannel of the plurality of microchannels comprises at least one microchannel branch which extends in a substantially radial direction and leads to a respective one said detection area.

### Claim 5 (depends on 1)

The microfluidic device according to claim 1, wherein each microchannel of the plurality of microchannels has associated at least two electrodes, the at least two electrodes being at one said detection area or in a position that is comprised between one said detection area and one of the first end and the second end of the corresponding microchannel of the plurality of microchannels.

### Claim 6 (depends on 1)

The microfluidic device according to claim 1, wherein each microchannel of the plurality of microchannels:
has a width of between 5 and 200 μm, and/or
has a depth or height of between 2 and 100 μm.

### Claim 7 (depends on 1)

The microfluidic device according to claim 1, wherein:
each microchannel of the plurality of microchannels has at least one surface portion defined by at least one of a hydrophilic material or a hydrophobic material, the hydrophilic material or the hydrophobic material belonging to at least one of the substrate and a covering element of the device that extends at least partially over each microchannel of the plurality of microchannels.

### Claim 8 (depends on 1)

The microfluidic device according to claim 1, wherein the substrate:
is configured for being mounted on a rotary member of a centrifugation device; and/or
is substantially disk-shaped.

### Claim 9 (depends on 1)

The microfluidic device according to claim 1, wherein defined in said surface of the substrate are a plurality of said microfluidic arrangements arranged substantially aligned with one another in the radial direction and/or set substantially according to a circumference.

### Claim 10 (depends on 1)

The microfluidic device according to claim 1, wherein said second distances are the same as one another.

### Claim 11 (depends on 1)

A method for the detection of particles possibly present in a fluid sample, comprising the steps of:
providing the microfluidic device according to claim 1;
introducing a volume of the fluid sample into each microchannel of the at least one microfluidic arrangement of the microfluidic device;
causing a rotation of the microfluidic device with respect to the corresponding centre of rotation at a centrifugation speed; and
detecting particles that have accumulated in the area for particle accumulation of each microchannel, in an optical and/or electrical way.

### Claim 12 (depends on 1)

A method for carrying out an antibiogram, comprising:
providing the microfluidic device according to claim 1;
providing a liquid medium containing micro-organisms, or microbes, or bacteria of at least one bacterial strain;
introducing a volume of the liquid medium into the plurality of first microchannels of the at least one first microfluidic arrangement of the microfluidic device;
causing an angular movement of the microfluidic device with respect to the corresponding centre of rotation at a centrifugation speed, via a centrifugation device; and
quantifying the number of micro-organisms, or microbes, or bacteria that have accumulated in the area for particle accumulation of each microchannel.

### Claim 13 (depends on 12)

The method according to claim 12, comprising:
i) pre-treating said first microchannels with at least one first antibiotic;
ii) obtaining a mass of the micro-organisms, or the microbes, or the bacteria;
iii) inoculating at least one part of said mass into the liquid medium;
iv) introducing the volume of the liquid medium into said first microchannels;
v) waiting for a period of time comprised between 10 min and 6 h;
vi) subjecting the microfluidic device to centrifugation; and
vii) quantifying the number of micro-organisms, or microbes, or bacteria that have accumulated in the area for particle accumulation of said first microchannels, carrying out a relative quantification between said first microchannels and second microchannels of the microfluidic device that have not been pre-treated with the at least one first antibiotic, in order to obtain a susceptibility profile of said micro-organisms, or microbes, or bacteria to the at least one first antibiotic.

### Claim 14 (depends on 13)

The method according to claim 13, wherein said mass referred to in step ii) is taken from a culture dish or else is obtained from a sample taken from a host organism.

### Claim 15

A microfluidic device for concentrating and detecting particles contained in a fluid sample, the microfluidic device comprising:
a substrate configured for being set in rotation with respect to a centre of rotation, the substrate having a surface at which at least one microfluidic arrangement is defined, which extends substantially according to a plane identified by the substrate, wherein the at least one microfluidic arrangement comprises a plurality of microchannels each having a first end and a second end, wherein:
each microchannel of the plurality of microchannels comprises, in a region thereof intermediate to the first end and the second end, at least one detection area that is at a first distance in a radial direction with respect to the centre of rotation of the substrate;
the first end and the second end of each microchannel of the plurality of microchannels are at second distances in the radial direction with respect to the centre of rotation of the substrate;
the first distance in the radial direction is greater than the second distances in the radial direction,
wherein the at least one detection area is an area for particle accumulation;
wherein the at least one microfluidic arrangement comprises at least one first duct or chamber and a second duct or chamber, the microchannels of the plurality of microchannels having the respective first ends and second ends connected in parallel to the first duct or chamber and to the second duct or chamber, respectively, one of the first duct or chamber and second duct or chamber being pre-arranged for receiving the fluid sample, such that the fluid sample can penetrate from one end of the microchannels of the plurality of microchannels, and air contained therein can progressively be vented at the other one of the first duct or chamber and second duct or chamber,
wherein the microchannels of the plurality of microchannels are the same as one another and extend at least in part substantially parallel to, or equidistant from, one another in the radial direction,
wherein, in the at least one detection area for particle accumulation, each microchannel of the plurality of microchannels is substantially V-shaped or U-shaped, or comprises two substantially convergent microchannel branches, the at least one detection area of each microchannel of the plurality of microchannels being free of exit ways for the fluid sample,
in such a way that particles contained in a volume of fluid of the fluid sample that penetrates into at least one microchannel of the plurality of microchannels concentrate in the at least one detection area as a result of the centrifugal force caused by a rotation of the substrate about the centre of rotation.