KR20180090597A - Cell electrical impedance spectroscopy measurement apparatus and method - Google Patents

Abstract

A cell impedance spectroscopy measurement apparatus comprises: a microfluidic channel providing a space for a flow of a fluid including a cell and having a channel size of a diameter of the cell or less; at least one pair of electrodes including a first electrode and a second electrode separately arranged so as to be in contact with the cell passing through the microfluidic channel within the microfluidic channel; and an electrical signal detector for supplying electrical signals having different frequencies by location of the cell passing through the microfluidic channel and detecting the electrical signal from the cell.

Description

TECHNICAL FIELD [0001] The present invention relates to a cell impedance spectroscopy measurement device,

The present invention relates to an apparatus and method for measuring cell impedance spectroscopy. More particularly, the present invention relates to a measuring device capable of measuring electrical characteristics of cells using impedance spectroscopy, and a method of measuring the cell impedance spectroscopy using the same.

Research is underway to determine the pathological state of cancer cells through the measurement of cellular electrical properties. Conventional impedance flow cytometry is suitable for counting and classification of cells but it is not sensitive to the measurement of cells without contact with electrodes and is not suitable for the quantification of metastatic cancer cells. To improve this, devices for measuring metastatic cancer cells with high sensitivity by directly contacting the cells with electrodes are being developed.

Micro-impedance spectroscopy is a device that directly measures electrodes in contact with cells. It measures the dielectric properties of cells according to frequency, and determines cell membrane capacitance, cell membrane resistance, cytoplasmic resistance, cytoplasmic capacitance, And permittivity can be measured. However, since the existing device measures electric signals of various frequencies at one electrode after performing cell trapping, additional devices for cell collection and operation time for cell collection are required.

An object of the present invention is to provide a device for measuring cell impedance spectroscopy capable of precisely measuring metastatic cancer cells such as cancer cells and reducing the measurement time without additional devices for cell collection of conventional micro impedance devices.

Another object of the present invention is to provide a method of measuring the cell impedance spectroscopy using the above-described apparatus for measuring cell impedance spectroscopy.

According to an aspect of the present invention, there is provided an apparatus for measuring cell impedance spectroscopy, comprising: a microfluidic channel having a channel size of less than a diameter of a cell, At least one electrode pair having a first electrode and a second electrode spaced apart from each other to be in contact with the cell passing through the microfluidic channel in the microfluidic channel, And an electrical signal detector for applying electrical signals of a frequency to the electrode pair and detecting an electrical signal from the cell.

In exemplary embodiments, a plurality of the electrode pairs may be provided, and the plurality of electrode pairs may be spaced apart from each other along the longitudinal direction of the microfluidic channel.

In exemplary embodiments, the electrical signal detector may include a plurality of detectors coupled to each of the plurality of electrode pairs for applying electrical signals of different frequencies, respectively.

In exemplary embodiments, the first and second electrodes of the first electrode pair among the electrode pairs have a first length, and the first and second electrodes of the second electrode pair, Lt; RTI ID = 0.0 > 1 < / RTI > length.

In exemplary embodiments, the electrode pair is provided, and the first and second electrodes may extend in the longitudinal direction of the microfluidic channel, respectively.

In exemplary embodiments, the electrical signal detector detects an electrical signal of the cell by applying a first frequency when the cell passes through the first location of the microfluidic channel, It is possible to detect an electrical signal of the cell by applying a second frequency different from the first frequency.

In exemplary embodiments, the first electrode and the second electrode may be disposed on first and second opposing sidewalls of the microfluidic channel, respectively.

In exemplary embodiments, at least one side wall of the microfluidic channel may be inclined or curved at an angle to the bottom surface.

In exemplary embodiments, the device for measuring cell impedance spectroscopy may further include a variable thin film structure constituting a part of a sidewall of the microfluidic channel.

In exemplary embodiments, the device for measuring cell impedance spectroscopy may further comprise an additional structure disposed in the microfluidic channel and for controlling an area through which the fluid passes.

In exemplary embodiments, the cell impedance spectrometer may further include a biochemical material film coated on one side wall of the microfluidic channel or on the first and second electrodes.

In exemplary embodiments, a plurality of the microfluidic channels may be arranged in series or parallel to each other, and the electrode pairs may be disposed in each of the microfluidic channels.

In exemplary embodiments, the channel sizes of the plurality of microfluidic channels may be equal to or different from each other.

According to another aspect of the present invention, there is provided a method for measuring a cell impedance spectroscopy, comprising the steps of: measuring at least one cell having a first electrode and a second electrode spaced apart from each other in a microfluidic channel having a first channel size; Place the electrode pairs. And a fluid containing cells having a diameter larger than the first channel size is introduced into the microfluidic channel. And an electrical signal of different frequency is applied to the electrode pair according to the position of the cell passing through the microfluidic channel. And an electrical signal is detected from the cell. And discharges the fluid from the microfluidic channel.

In exemplary embodiments, disposing the electrode pair may include disposing a plurality of electrode pairs spaced apart along the longitudinal direction of the microfluidic channel, wherein applying the electrical signal to the electrode pair And applying electrical signals of different frequencies to each of the plurality of electrode pairs.

In the exemplary embodiments, disposing the electrode pair may include disposing the first and second electrodes to extend in the longitudinal direction of the microfluidic channel, respectively, wherein the electrical signal is applied to the electrode pair The application of an electrical signal of a first frequency when the cell passes through the first location of the microfluidic channel and the application of an electrical signal of a second frequency different from the first frequency when the cell passes the second location of the microfluidic channel. Lt; RTI ID = 0.0 > frequency. ≪ / RTI >

In the cell impedance spectroscopic measurement apparatus and method according to the present invention, electrical signals of different frequencies are applied to at least one pair of electrodes provided in a microfluidic channel having a channel size (width or depth) And the electrical characteristics of the cells can be measured.

By measuring the dielectric properties of the cells by detecting electrical signals at different frequencies, it is possible to quantify metastatic cancer cells and complex and heterogeneous cells, and to produce channel cross-sections of the microfluidic channels below the cell diameter, Since there is no need for a device, no operation time is required for cell collection, and measurement is possible in a short time.

However, the effects of the present invention are not limited to the above-mentioned effects, and may be variously expanded without departing from the spirit and scope of the present invention.

1 is an exploded perspective view showing a device for measuring cell impedance spectroscopy according to exemplary embodiments.
2 is a plan view showing the cell impedance spectroscopy measuring apparatus of FIG.
3 is a plan view showing electrode pairs disposed in a microfluidic channel of the device for measuring cell impedance spectroscopy of FIG.
4 is a cross-sectional view taken along line AA 'of FIG.
5 is a block diagram showing an electrical signal detector of the apparatus for measuring cell impedance spectroscopy of FIG.
6 is a plan view showing a cell impedance spectroscopy measurement device according to exemplary embodiments.
7 is a cross-sectional view taken along line BB 'of FIG.
8 is a block diagram showing an electrical signal detector of the cell impedance spectroscopy measurement apparatus of FIG.
9 is a cross-sectional view illustrating a microfluidic channel in accordance with exemplary embodiments.
10 is a cross-sectional view illustrating a microfluidic channel in accordance with exemplary embodiments.
11A-11F are cross-sectional views illustrating electrodes of a pair of electrodes in a microfluidic channel in accordance with exemplary embodiments.
12A-C are cross-sectional views illustrating microfluidic channels in accordance with exemplary embodiments.
13 is a plan view showing a device for measuring cell impedance spectroscopy according to exemplary embodiments.
14 is a plan view showing a microfluidic channel in accordance with exemplary embodiments.
15 is a plan view of a device for measuring cell impedance spectroscopy according to exemplary embodiments.
16 is a top view of a device for measuring cell impedance spectroscopy according to exemplary embodiments.
17 is a flow chart illustrating a method for measuring cell impedance spectroscopy according to exemplary embodiments.
18 is a circuit diagram showing an electrical equivalent circuit model when cells are in a microfluidic channel.
FIGS. 19 and 20 are graphs showing electrical characteristics of cells measured by a cell impedance spectroscopy measuring apparatus according to an embodiment.

For the embodiments of the invention disclosed herein, specific structural and functional descriptions are set forth for the purpose of describing an embodiment of the invention only, and it is to be understood that the embodiments of the invention may be practiced in various forms, The present invention should not be construed as limited to the embodiments described in Figs.

The present invention is capable of various modifications and various forms, and specific embodiments are illustrated in the drawings and described in detail in the text. It is to be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but on the contrary, is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms may be used for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the first component may be referred to as a second component, and similarly, the second component may also be referred to as a first component.

It is to be understood that when an element is referred to as being "connected" or "connected" to another element, it may be directly connected or connected to the other element, . On the other hand, when an element is referred to as being "directly connected" or "directly connected" to another element, it should be understood that there are no other elements in between. Other expressions that describe the relationship between components, such as "between" and "between" or "neighboring to" and "directly adjacent to" should be interpreted as well.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprise", "having", and the like are intended to specify the presence of stated features, integers, steps, operations, elements, components, or combinations thereof, , Steps, operations, components, parts, or combinations thereof, as a matter of principle.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be construed as meaning consistent with meaning in the context of the relevant art and are not to be construed as ideal or overly formal in meaning unless expressly defined in the present application .

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The same reference numerals are used for the same constituent elements in the drawings and redundant explanations for the same constituent elements are omitted.

1 is an exploded perspective view showing a device for measuring cell impedance spectroscopy according to exemplary embodiments. 2 is a plan view showing the cell impedance spectroscopy measuring apparatus of FIG. 3 is a plan view showing electrode pairs disposed in a microfluidic channel of the device for measuring cell impedance spectroscopy of FIG. 4 is a cross-sectional view taken along the line A-A 'in FIG. 5 is a block diagram showing an electrical signal detector of the apparatus for measuring cell impedance spectroscopy of FIG.

1 to 5, the apparatus for measuring cell impedance spectroscopy 10 includes a flow path 110 having a microfluidic channel 112 through which a fluid including a cell C flows, An electrode pair, and an electrical signal detector for applying an electrical signal to the electrode pair and detecting an electrical signal from the cell (C).

In the exemplary embodiments, the flow path 110 may include an inlet portion 150 and an outlet portion 160 that are provided at both sides, respectively. The flow path 110 can provide a space for the flow of the fluid including microparticles such as the cells (C). The fluid may flow into the microfluidic channel 112 of the flow path 110 through the inlet 150 and may be discharged through the outlet 160.

The flow direction and the flow rate of the fluid in the microfluidic channel 112 of the flow path 110 can be adjusted. This provides the appropriate flow rate and flow direction for each cell or cell characteristic. For example, at least one of the inflow section 150 and the outflow section 160 may include a fluid supply element (not shown) to adjust the direction and flow rate of the fluid in the microfluidic channel 112. The fluid supply element may include mechanical devices (syringe pumps, pneumatic membrane pumps, vibrating membrane pumps), electrical or magnetic devices (electrohydrodynamic pumps, magnetorheological pumps), thermodynamic devices, and the like.

The flow path 110 adjacent to the inflow section 150 and the outflow section 160 may further include a guiding structure for guiding the flow of the fluid. The guiding structure may have an inclined surface for guiding fluid flow to the microfluidic channel 112. Accordingly, it is possible to adjust the time during which the cells pass through the inflow section 150 and the outflow section 160.

The fluid may be a solution comprising biochemical microparticles. Examples of the solution may be blood, body fluid, cerebrospinal fluid, urine, sputum or a mixture or diluted solution thereof. Examples of the fine particles include blood cancer cells, stem cells, mesenchymal cells, interstitial cells, and the like.

The device for measuring cell impedance spectroscopy can be formed by a micro fabrication technique including photolithography, soft lithography, and the like. The microfluidic channel may be formed using a polymer material, an inorganic material, or the like. Examples of the polymer material include PDMS, PMMA, SU-8, and the like. Examples of the inorganic material include glass, quartz, silicon and the like. The electrode pair may be formed using a metal material. Examples of the metal material include gold, silver, platinum, copper and aluminum.

1 to 4, the microparticle analyzer 10 may include a first substrate 100, a second substrate 102, and a third substrate 104 which are sequentially stacked .

At least one electrode pattern may be formed on the first substrate 100. Specifically, the first to fourth electrode patterns 300, 302, 304, and 306 may be spaced apart from each other along the first direction (X direction). The first electrode pattern 300 may include a first pattern 300a and a second pattern 300b spaced from each other with a center line of the first substrate 100 interposed therebetween. The second electrode pattern 302 may include a first pattern 302a and a second pattern 302b spaced apart from each other with a center line of the first substrate 100 therebetween. The third electrode pattern 304 may include a first pattern 304a and a second pattern 304b spaced apart from each other with a center line of the first substrate 100 therebetween. The fourth electrode pattern 306 may include a first pattern 306a and a second pattern 306b spaced apart from each other with a center line of the first substrate 100 therebetween. As will be described later, one end of the first and second patterns may include a connection terminal for connection with the electrical signal detector.

The second substrate 102 may be formed on the first substrate 100 to define the flow path 110 having the microfluidic channel 112. The flow path 110 may extend in the first direction (X direction). The third substrate 104 may be formed on the second substrate 102. The third substrate 104 may include an inlet 150 and an outlet 160 connected to both ends of the flow path 110.

The first pattern 300a and the second pattern 300b of the first electrode pattern 300 may extend in opposite directions from the microfluidic channel 112. The first pattern 300a and the second pattern 300b may extend in a second direction (Y direction) perpendicular to the first direction. The first pattern 300a includes a first electrode 310a disposed in the microfluidic channel 112, a first connection terminal 320a spaced apart from the microfluidic channel 112, and a first connection 310a extending from the first electrode 310a. And a first connection pattern 330a extending to the terminal 320a. Similarly, the second pattern 300b includes a second electrode 310b disposed in the microfluidic channel 112, a second connection terminal 320b spaced from the microfluidic channel 112, and a second electrode 310b. And a second connection pattern 330b extending from the second connection terminal 320b to the second connection terminal 320b.

The first pattern 302a and the second pattern 302b of the second electrode pattern 302 may extend in opposite directions from the microfluidic channel 112. [ The first pattern 302a and the second pattern 302b may extend in a second direction (Y direction) orthogonal to the first direction. The first pattern 302a is disposed in the microfluidic channel 112 A first connection terminal 322a spaced from the microfluidic channel 112 and a first connection pattern 332a extending from the first electrode 312a to the first connection terminal 322a . Similarly, the second pattern 302b includes a second electrode 312b disposed in the microfluidic channel 112, a second connection terminal 322b spaced from the microfluidic channel 112, and a second electrode 312b. And a second connection pattern 332b extending from the second connection terminal 322b to the second connection terminal 322b.

The first pattern 304a and the second pattern 304b of the third electrode pattern 304 may extend in opposite directions from the microfluidic channel 112. The first pattern 304a and the second pattern 304b may extend in a second direction (Y direction) perpendicular to the first direction. The first pattern 304a includes a first electrode 314a disposed in the microfluidic channel 112, a first connection terminal 324a spaced from the microfluidic channel 112, And a first connection pattern 334a extending to the terminal 324a. Similarly, the second pattern 304b includes a second electrode 314b disposed in the microfluidic channel 112, a second connection terminal 324b spaced from the microfluidic channel 112, and a second electrode 314b. And a second connection pattern 334b extending from the second connection terminal 324b to the second connection terminal 324b.

The first pattern 306a and the second pattern 306b of the fourth electrode pattern 306 may extend in opposite directions from the microfluidic channel 112. The first pattern 306a and the second pattern 306b may extend in a second direction (Y direction) perpendicular to the first direction. The first pattern 306a includes a first electrode 316a disposed in the microfluidic channel 112, a first connection terminal 326a spaced apart from the microfluidic channel 112, and a first connection 316b extending from the first electrode 316a. And a first connection pattern 336a extending to the terminal 326a. Similarly, the second pattern 306b includes a second electrode 316b disposed in the microfluidic channel 112, a second connection terminal 326b spaced from the microfluidic channel 112, and a second electrode 316b. And a second connection pattern 336b extending from the second connection terminal 326b to the second connection terminal 326b.

In the exemplary embodiments, a plurality of first through fourth electrode pairs may be spaced apart from each other along the longitudinal direction (X direction) of the microfluidic channel 112 in the microfluidic channel 112. The first electrode pair may include a first electrode 310a and a second electrode 310b of the first electrode pattern 300. The second electrode pair may include a first electrode 312a and a second electrode 312b of the second electrode pattern 302. The third electrode pair may include a first electrode 314a and a second electrode 314b of the third electrode pattern 304. [ The fourth electrode pair may include a first electrode 316a and a second electrode 316b of the fourth electrode pattern 306. [ The first pair of electrodes may be disposed at a first position from an inlet in the microfluidic channel 112. The second pair of electrodes may be disposed at a second location further away from the first location from the inlet of the microfluidic channel 112. The third pair of electrodes may be disposed at a third location further away from the second location from the inlet of the microfluidic channel 112. The fourth pair of electrodes may be disposed at a fourth location further away from the third location from the inlet of the microfluidic channel 112.

As shown in FIGS. 3 and 4, the microfluidic channel 112 may have a channel size D2 that is less than the diameter D1 of the cell C. Here, the channel size may represent a dimension such as a width, a depth, a diameter, and the like. The first electrode 310a and the second electrode 310b of the first electrode pair may be disposed on the first sidewall 114a and the second sidewall 114b facing each other of the microfluidic channel 112 . Therefore, when the cells C in the fluid pass through the microfluidic channel 112, different sites of the cells C may contact the first electrode 310a and the second electrode 310b, respectively.

For example, the cell (C) may have a diameter of 1 탆 to 100 탆. The microfluidic channel 112 may have a channel size (width or depth) ranging from 20% to 50% of the diameter of the cell (C). When the cells (C) are two kinds of breast cancer cell lines (MCF7, MDA 231), the average diameters of the cells (C) are 17.26 탆 and 17.85 탆, and the channel size (width or depth) And may be 4 탆 to 10 탆.

As shown in FIG. 5, the electrical signal detector may apply electrical signals of different frequencies to the electrode pairs according to positions in the microfluidic channel 112 of the cell passing through the microfluidic channel 112 . Specifically, the electrical signal detector includes first, second, third and fourth detectors connected to each of the first, second, third and fourth electrode pairs for applying electrical signals of different frequencies, (400, 402, 404, 406).

The first detector 400 may be connected to the first connection terminal 320a and the second connection terminal 320b of the first electrode pattern 300. [ The AC signal having the first frequency range may be applied to the first connection terminal 320a and the second connection terminal 320b may be grounded. For example, the first detector 400 may apply an electrical signal having a frequency of 100 kHz to the first electrode pair.

The second detector 402 may be connected to the first connection terminal 322a and the second connection terminal 322b of the second electrode pattern 302. [ The AC signal having the second frequency range may be applied to the first connection terminal 322a and the second connection terminal 322b may be grounded. For example, the second detector 402 may apply an electrical signal having a frequency of 500 kHz to the second electrode pair.

The third detector 404 may be connected to the first connection terminal 324a and the second connection terminal 324b of the third electrode pattern 304. [ The AC signal having the third frequency range may be applied to the first connection terminal 324a and the second connection terminal 324b may be grounded. For example, the third detector 404 may apply an electrical signal having a frequency of 900 kHz to the third electrode pair.

The fourth detector 406 may be connected to the first connection terminal 326a and the second connection terminal 326b of the fourth electrode pattern 306. [ The AC signal having the fourth frequency range may be applied to the first connection terminal 326a and the second connection terminal 326b may be grounded. For example, the fourth detector 406 may apply an electrical signal having a frequency of 1.3 MHz to the fourth electrode pair.

The frequencies applied to the first through fourth electrode pairs may be regularly increased or decreased. Alternatively, the frequencies applied to the first through fourth electrode pairs may have irregularly different values. A plurality of electrical signals having the most significant frequency can be selected according to the characteristics of each cell and the characteristics thereof.

The first to fourth detectors 400, 402, 404, and 406 can detect the magnitude and phase of the electrical signal according to the change in frequency. By detecting the electrical characteristics of the cell (C) (cell membrane, cytoplasm), cancer cells can be discriminated and quantified.

As described above, the cell impedance spectroscopy measuring apparatus 10 is configured to apply electric signals of different frequencies to a plurality of electrode pairs disposed in the microfluidic channel 112 having a diameter smaller than the diameter of the cell C, (C) can be measured.

When the cell (C) is measured by an electrical signal of a different frequency, it is possible to obtain the dielectric characteristics of the cells (C) different from each other at a frequency, thereby precisely quantifying cells having complex and heterogeneous characteristics such as metastatic cancer cells have.

Since the electrode pair is in direct contact with the cell C, it is highly sensitive in the measurement of cell characteristics, and since an electric signal of a different frequency is measured in a plurality of electrode pairs rather than one electrode pair, an additional device for cell collection is not required It is possible to measure in a short time because no operation time is required for cell collection.

6 is a plan view showing a cell impedance spectroscopy measurement device according to exemplary embodiments. 7 is a cross-sectional view taken along line B-B 'of FIG. 8 is a block diagram showing an electrical signal detector of the cell impedance spectroscopy measurement apparatus of FIG. The cell impedance spectroscopy measuring device is substantially the same as or similar to the cell impedance spectroscopy measuring device described in Figs. 1 to 5 except for the configuration of the electrode pair and the electric signal detector. Accordingly, the same constituent elements are denoted by the same reference numerals, and repetitive description of the same constituent elements is omitted.

6 to 8, the apparatus for measuring a cell impedance spectroscopy 11 includes a pair of electrodes disposed in a microfluidic channel 112 and a plurality of microfluidic channels 112 disposed in the microfluidic channel 112 And an electrical signal detector for applying electrical signals of different frequencies to the electrode pair and detecting an electrical signal from the cell.

In the exemplary embodiments, one electrode pair may be disposed in the microfluidic channel 112. The first electrode pattern 300 may include a first pattern 300a and a second pattern 300b spaced apart from each other with a center line of the microfluidic channel 112 interposed therebetween. The first pattern 300a and the second pattern 300b of the first electrode pattern 300 may extend in opposite directions from the microfluidic channel 112. The first pattern 300a and the second pattern 300b may extend in a second direction (Y direction) orthogonal to the first direction (X direction).

The first pattern 300a includes a first electrode 310a disposed in the microfluidic channel 112, a first connection terminal 320a spaced apart from the microfluidic channel 112, and a first connection 310a extending from the first electrode 310a. And a first connection pattern 330a extending to the terminal 320a. Similarly, the second pattern 300b includes a second electrode 310b disposed in the microfluidic channel 112, a second connection terminal 320b spaced from the microfluidic channel 112, and a second electrode 310b. And a second connection pattern 330b extending from the second connection terminal 320b to the second connection terminal 320b. The first electrode 310a and the second electrode 310b of the first electrode pattern 300 may constitute the first electrode pair.

The first electrode 310a and the second electrode 310b may extend in the longitudinal direction of the microfluidic channel 112. The first and second electrodes 310a and 310b may have a length of several tens to several tens of times the diameter of the cells. The first electrode 310a may cover the first sidewall 114a as a whole and the second electrode 310b may cover the second sidewall 114b as a whole. The first electrode 310a and the second electrode 310b may be spaced apart from each other by an interval smaller than the diameter of the cell. The first and second electrodes 310a and 310b can directly form at least a part of the microfluidic channel through which the fluid flows. Therefore, the cells flowing into the microfluidic channel 112 can move while being in direct contact with the first electrode 310a and the second electrode 310b.

The electrical signal detector may include a detector 400 for applying an alternating signal of variable frequency to the first and second electrodes 310a and 310b of the electrode pair. The detector 400 detects an electrical signal of the cell by applying a first frequency when the cell passes through the first location of the microfluidic channel 112 and when the cell detects the second location of the microfluidic channel 112 It is possible to detect an electrical signal of the cell by applying a second frequency different from the first frequency. The detector 400 can detect the electrical signals of the cells by applying different frequencies at certain intervals during the movement of the cells in the microfluidic channel 112.

Therefore, when the cells move between the first and second electrodes 310a and 310b in the microfluidic channel 112 in direct contact therewith, An electrical signal of a different frequency can be applied and the electrical characteristics of the cell can be detected.

9 is a cross-sectional view illustrating a microfluidic channel in accordance with exemplary embodiments.

9, a cell impedance spectroscopy apparatus includes a biochemical material film 140 (not shown) coated on at least one side of a pair of electrodes 310a, 310b in at least one side wall of a microfluidic channel 112 or a microchannel channel 112, ). ≪ / RTI > The biochemical material membrane 140 may be formed in the micro-rapeseed channel 112 to increase or prevent adhesion to the cell. The biochemical material membrane 140 may comprise an antibody. Examples of the antibody include an anti-EpCAM antibody and an anti-CK antibody.

10 is a cross-sectional view illustrating a microfluidic channel in accordance with exemplary embodiments.

Referring to FIG. 10, at least one side wall of the microfluidic channel 112 may be inclined or curved at an angle with respect to the bottom surface. The first sidewall 114a and the second sidewall 114b facing each other may have a vertical cross-sectional profile of a concave shape along the vertical direction.

11A-11F are cross-sectional views illustrating electrodes of a pair of electrodes in a microfluidic channel in accordance with exemplary embodiments.

Referring to FIGS. 11A to 11, the electrode pairs disposed in the microfluidic channel may include a first electrode 310a and a second electrode 310b spaced from each other. The first electrode 310a and the second electrode 310b may be disposed symmetrically or asymmetrically with each other in the microfluidic channel. The first and second electrodes 310a and 310b may be disposed at the upper, middle, or lower portion of the microfluidic channel.

Also, two or more first electrodes 310a and two or more second electrodes 310b may be disposed in the microfluidic channel. Through this, the electrical signals of cells can be measured three-dimensionally.

12A-C are cross-sectional views illustrating microfluidic channels in accordance with exemplary embodiments.

12A to 12C, the apparatus for measuring cell impedance spectroscopy may further include a variable thin film structure 520, 522 constituting a part of a sidewall of a microfluidic channel.

12A, the first variable thin film 500 is sandwiched between the second substrate 102 and the third substrate 104 and includes a first variable thin film structure 520, which is a part of the first variable thin film 500 May constitute at least one side wall of the microfluidic channel.

A recess may be formed on the lower surface of the third substrate 104 at a position corresponding to the first variable thin film structure 520. The first variable thin film 500 may cover the recess to form the first thin film control line 510. The first thin film control line 510 may be connected to an external hydraulic or pneumatic source to deform the first deformable thin film structure 520.

Accordingly, the first deformable thin film structure 520 may be deformed by an external force to change the size of the microfluidic channel or to pressurize the cells.

12B, the second variable thin film 502 is interposed between the first substrate 100 and the second substrate 102, and the second variable thin film structure 522 (a part of the second variable thin film 502) May constitute at least one side wall of the microfluidic channel.

A recess may be formed on the upper surface of the first substrate 100 at a position corresponding to the second variable thin film structure 522. The second variable thin film 502 may cover the recess to form a second thin film control line 512. The second thin film control line 512 may be connected to an external hydraulic or pneumatic source to deform the second deformable thin film structure 522.

Accordingly, the second variable thin film structure 522 may be deformed by an external force to change the size of the microfluidic channel or to pressurize the cells.

12C, the first deformable thin film structure 520 constitutes at least a portion of the upper sidewall of the microfluidic channel and the second deformable thin film structure 522 forms at least a portion of the lower sidewall of the microfluidic channel .

Accordingly, the first and second variable thin film structures 520 and 522 may be deformed by an external force to change the size of the microfluidic channel or pressurize the cells.

13 is a plan view showing a device for measuring cell impedance spectroscopy according to exemplary embodiments.

Referring to FIG. 13, the apparatus for measuring cell impedance spectroscopy may further include an additional structure 600 disposed in the microfluidic channel 112 for controlling an area through which the fluid passes.

A plurality of additional structures 600 may be disposed behind each pair of electrodes. The additional structure may increase the contact time between the cell and the electrode pair as a stopper.

14 is a plan view showing a microfluidic channel in accordance with exemplary embodiments.

Referring to FIG. 14, the electrode lengths of the electrode pairs may be the same or different. Specifically, the first and second electrodes 310a and 310b of the first electrode pair may have a first length L1 in the longitudinal direction of the microfluidic channel 112. The first and second electrodes 312a and 312b of the second electrode pair may have a second length L2 greater than the first length L1 in the longitudinal direction of the microfluidic channel 112. [ The first and second electrodes 314a and 314b of the third electrode pair may have a third length L3 that is greater than the second length L2 in the longitudinal direction of the microfluidic channel 112. [

15 is a plan view of a device for measuring cell impedance spectroscopy according to exemplary embodiments.

Referring to FIG. 15, the apparatus for measuring cell impedance spectroscopy may include a plurality of first to third microfluidic channels 112, 122 and 132 arranged in series with each other.

The first to third microfluidic channels 112, 122, 132 may have the same or different diameters. Specifically, the first microfluidic channel 112 has a first diameter D2, the second microfluidic channel 122 has a second diameter D3 that is larger than the first diameter D2, The fluid channel 132 may have a fourth diameter D4 that is greater than the second diameter D3.

The cell impedance spectroscopy apparatus includes a first group of electrode pairs 20 disposed in a first microfluidic channel 112, a second group of electrode pairs 22 disposed in a second microfluidic channel 122, And a third group of electrode pairs (24) disposed in the third fine rapid channel (132).

Therefore, it is possible to additionally measure the mechanical properties of cells such as cell passage time and cell size change.

16 is a top view of a device for measuring cell impedance spectroscopy according to exemplary embodiments.

Referring to FIG. 16, the apparatus for measuring cell impedance spectroscopy may include a plurality of first to third microfluidic channels 112, 122, 132 arranged in parallel with each other.

The first to third microfluidic channels 112, 122, 132 may have the same or different diameters. Specifically, the first microfluidic channel 112 has a first diameter D2, the second microfluidic channel 122 has a second diameter D3 that is larger than the first diameter D2, The fluid channel 132 may have a fourth diameter D4 that is greater than the second diameter D3.

The device for measuring cell impedance spectroscopy includes a first group of electrode pairs 30 disposed in a first microfluidic channel 112, a second group of electrode pairs 32 disposed in a second microfluidic channel 122, And a third group of electrode pairs 34 disposed in the third microchannel channel 132.

Therefore, it is possible to additionally measure the mechanical properties of cells such as cell passage time and cell size change.

Hereinafter, a method for measuring the electrical characteristics of a cell using the above-described cell impedance spectroscopic measurement apparatus will be described.

17 is a flow chart illustrating a method for measuring cell impedance spectroscopy according to exemplary embodiments.

Referring to FIG. 17, a microfluidic channel having a first diameter and at least one pair of electrodes is provided, and a fluid including cells C is introduced into the microfluidic channel through an inlet 150 (S100 ). At this time, the cells C may have a diameter larger than the first diameter.

A plurality of electrode pairs may be disposed along the longitudinal direction of the microfluidic channel 112 in the microfluidic channel 112 of the measurement apparatus 10 of FIG. One electrode pair may be disposed in the microfluidic channel 112 of the measuring device 11 of FIG.

Next, an electrical signal having a frequency different according to the position of the cell passing through the microfluidic channel is applied to the electrode pair (S110), and an electrical signal of the cell is detected (S120). Thereafter, the fluid is discharged from the microfluidic channel (S130).

In the case of the measuring apparatus 10 of FIG. 1, electrical signals of different frequencies may be applied to each of the plurality of electrode pairs. 6, when the cell passes through the first position of the microfluidic channel 112, an electrical signal of a first frequency is applied, and after the cell passes through the first position, And may apply an electrical signal of a second frequency that is different from the first frequency when passing through the second position of the second electrode 112. The electrical signal detector can detect the magnitude and the phase of the electrical signal according to the frequency change.

18 is a circuit diagram showing an electrical equivalent circuit model when cells are in a microfluidic channel.

Referring to Fig. 18, in the equivalent circuit model of cells, Rm represents the resistance of the cell membrane, Re represents the resistance of the electric double layer, Rc represents the resistance of the cytoplasm, and Cm represents the capacitance of the cell membrane. The results obtained through the cell impedance spectroscopy can be applied to modeling to derive electrical properties of the cell membrane and cytoplasm.

FIGS. 19 and 20 are graphs showing electrical characteristics of cells measured by a cell impedance spectroscopy measuring apparatus according to an embodiment.

Referring to FIGS. 19 and 20, results are shown according to the size and phase of two types of breast cancer cell lines (MCF7, MDA 231) and a control group (PBS) according to changes in frequency. Through the electrical characteristics measured by the cell impedance spectroscopic measurement device, the resistance of the cell membrane, the resistance of the electric double layer, the resistance of the cytoplasm, and the capacitance of the cell membrane can be obtained, and the cancer cell can be discriminated and quantified.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined in the following claims. It can be understood that it is possible.

10, 11: Cell Impedance Spectrometer
100: first substrate 102: second substrate
104: third substrate 110:
112, 122, 132: microfluidic channel 140: biochemical material membrane
150: inlet 160: outlet
300, 302, 304, 306: first to fourth electrode patterns
300a, 302a, 304a, 306a: a first pattern
300b, 302b, 304b, and 306b: a second pattern
310a, 312a, 314a, 316a:
310b, 312b, 314b, 316b:
320a, 322a, 324a, 326a: a first connection terminal
320b, 322b, 324b, 326b: a second connection terminal
330a, 332a, 334a, 336a: a first connection pattern
330b, 332b, 334b, 336b: a second connection pattern
400, 402, 404, 406: first to fourth detectors
500: first variable thin film 502: second variable thin film
510: first thin film control line 512: second thin film control line
520: first variable thin film structure 522: second variable thin film structure
600: Stopper

Claims (16)

A microfluidic channel providing a space for fluid flow including cells and having a channel size below the diameter of the cells;
At least one electrode pair having a first electrode and a second electrode spaced apart from each other to be in contact with the cells passing through the microfluidic channel in the microfluidic channel; And
And an electrical signal detector for applying electrical signals of different frequencies to the electrode pair according to positions of the cells passing through the microfluidic channels and for detecting an electrical signal from the cells. The cell impedance spectrometer of claim 1, wherein the plurality of electrode pairs are disposed in the longitudinal direction of the microfluidic channel. 3. The apparatus according to claim 2, wherein the electrical signal detector includes a plurality of detectors connected to each of the plurality of electrode pairs for applying electrical signals of different frequencies. 3. The plasma display panel of claim 2, wherein the first and second electrodes of the first pair of electrodes have a first length and the first and second electrodes of the second pair of electrodes comprise a first length And a second length equal to or different from the second length. The apparatus of claim 1, wherein the electrode pair is provided with one electrode pair, and the first and second electrodes extend in the longitudinal direction of the microfluidic channel. 6. The method of claim 5, wherein the electrical signal detector detects an electrical signal of the cell by applying a first frequency when the cell passes through the first location of the microfluidic channel, And a second frequency different from the first frequency is applied to the cell to detect an electrical signal of the cell. The apparatus of claim 1, wherein the first electrode and the second electrode are respectively disposed on first and second sidewalls facing each other of the microfluidic channel. The apparatus according to claim 1, wherein at least one side wall of the microfluidic channel has an inclined or curved shape at a predetermined angle with respect to a bottom surface. The cell impedance spectroscopic measurement apparatus according to claim 1, further comprising a variable thin film structure constituting a part of a sidewall of the microfluidic channel. The apparatus of claim 1, further comprising an additional structure disposed within the microfluidic channel and for controlling an area through which the fluid passes. The apparatus of claim 1, further comprising a biochemical material film coated on one side wall of the microfluidic channel or on the first and second electrodes. The apparatus according to claim 1, wherein a plurality of the microfluidic channels are arranged in series or parallel to each other, and the electrode pairs are disposed in each of the microfluidic channels. The apparatus of claim 1, wherein the channel sizes of the plurality of microfluidic channels are equal to or different from each other. Disposing at least one electrode pair having a first electrode and a second electrode so as to be spaced apart from each other in a microfluidic channel having a first channel size;
Introducing a fluid including cells having a diameter larger than the first channel size into the microfluidic channel;
Applying electrical signals of different frequencies to the electrode pairs according to positions of the cells passing through the microfluidic channel;
Detecting an electrical signal from the cell; And
And discharging the fluid from the microfluidic channel. 15. The method of claim 14, wherein disposing the electrode pair comprises disposing a plurality of electrode pairs apart from each other along the longitudinal direction of the microfluidic channel,
Wherein applying the electrical signal to the electrode pair comprises applying electrical signals of different frequencies to each of the plurality of electrode pairs. 15. The method of claim 14, wherein disposing the electrode pair comprises disposing the first and second electrodes to extend in the longitudinal direction of the microfluidic channel, respectively,
Wherein applying the electrical signal to the electrode pair applies an electrical signal of a first frequency when the cell passes through the first location of the microfluidic channel and when the cell passes through the second location of the microfluidic channel And applying an electrical signal of a second frequency different from the first frequency.

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