KR102646803B1 - Apparatus and method for sensing capacitance of cell based on simultaneous optical tweezing - Google Patents

Abstract

An in situ cell capacitance sensing device and method for detecting capacitance for a single cell or an array of multiple cells based on simultaneous optical tweezing are disclosed. The simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention includes a substrate on which cells are placed, a pair of electrodes spaced apart on both sides of the substrate, and a capacitance between the pair of electrodes. A cell analysis device including an analyzer configured to measure and analyze the characteristics of the cell; an optical twizing device including a laser device that irradiates a focused laser beam for trapping, and configured to adjust the position of the cell with respect to the pair of electrodes by adjusting an irradiation position of the focused laser beam with respect to the substrate; and an image acquisition device configured to capture a cell image on the pair of electrodes and the substrate to obtain a cell image that can be used as a reference for adjusting the position of the cell.

Description

{Apparatus and method for sensing capacitance of cell based on simultaneous optical tweezing}

The present invention relates to a cell capacitance sensing device and method, and more specifically to an in situ cell capacitance sensing device and method for a single cell or an array of multiple cells based on simultaneous optical tweezing. It relates to a capacitance sensing device and method.

Technologies for measuring various cell activities with high sensitivity have been studied, and in particular, a biosensor that detects changes in capacitance caused by cells has been developed as one of the label-free real-time detection technologies. Normally, cells become polarized under an external electric field due to the polarization of surface charges that accumulate on the cell membrane. This polarization is sensitively altered by the electrical properties of cellular components and is proportional to the membrane potential at low electric fields. Real-time capacitive biosensors provide valuable and accurate information about cell activity by measuring changes in capacitance, and are being used to measure a variety of biological interactions, including real-time cell differentiation, cancer cell differentiation, and cell dynamics. In particular, unlike existing fluorescence imaging methods, using a capacitive sensor can provide quantitative information on the epidermal growth factor receptor, a transmembrane glycoprotein of the cell membrane, and can be applied to effectively detect cancer biomarkers.

In general, most cell activity measurements using capacitive sensors use an average analysis method of a group of cells because it is difficult to control the number of cells settling between electrodes. Recognizing the parameters of individual cells is of great importance for many applications, such as cell phenotyping, stem cell sorting, or cell delivery via endocytosis. Several single cell analysis methods have been developed, including patch clamping, alternating current (AC) electrokinetics, and microfluidics.

However, patch clamping generally has the disadvantage of being labor-intensive, difficult to apply, and slow in the serial process for cell analysis. Methods utilizing AC electrokinetics and microfluidics typically require modification of cell structure to fit cells into narrow capillaries. Moreover, single cell analysis in these cases is generally opportunistic and has limitations that do not allow precise control, selectivity or directional mobility of individual cells. Cells usually grow together, and although, especially in cases such as neurons, every molecule transported out of one cell affects the behavior of neighboring cells, studies of conventional cell capacitance sensors have not been able to pinpoint interactions between cells. There were limitations to the analysis.

The present invention is intended to provide an in situ cell capacitance sensing device and method for detecting the capacitance of a single cell or an array of multiple cells based on simultaneous optical tweezing.

Meanwhile, the technical problems to be achieved in the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned will be clearly apparent to those skilled in the art from the description below. It will be understandable.

The simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention includes a substrate on which cells are placed, a pair of electrodes spaced apart on both sides of the substrate, and a capacitance between the pair of electrodes. A cell analysis device including an analyzer configured to measure and analyze the characteristics of the cell; an optical twizing device including a laser device that irradiates a focused laser beam for trapping, and configured to adjust the position of the cell with respect to the pair of electrodes by adjusting an irradiation position of the focused laser beam with respect to the substrate; and an image acquisition device configured to capture a cell image on the pair of electrodes and the substrate to obtain a cell image that can be used as a reference for adjusting the position of the cell.

When a first cell array to be analyzed is selected from among a plurality of preset cell arrays, the optical twirling device selects one or more first cells from among a plurality of cells included in the cell image according to the first cell array; Measure a first coordinate corresponding to the position of the first cell in the cell image, and determine a target position for each first cell based on the first cell array and the first coordinate; It may be configured to adjust the position of the first cells to match the first cell arrangement by moving at least one of the substrate and the laser device according to the target position.

The plurality of cell arrays may include a single cell array, a horizontal double cell array, a vertical double cell array, a horizontal triple cell array, a vertical triple cell array, and a multi-matrix cell array with two or more cells arranged in each of the horizontal and vertical directions. You can.

The analyzer measures a first capacitance value by measuring impedance between the pair of electrodes while the cells are arranged in the first cell array on the substrate; Applying the first capacitance value to a correction function set for the first cell array to calculate a second capacitance value representing the capacitance of a single cell; It may be configured to analyze the characteristics of the cell based on the second capacitance value.

The image acquisition device includes a light source that outputs white light toward cells on the substrate to acquire images of the cells; a focusing lens provided on the optical path of the white light output from the light source to focus the white light toward the cells on the substrate; an objective lens provided on the optical path of the white light passing through the substrate to converge the white light; a color-selective mirror provided on the optical path of the white light condensed by the objective lens to reflect the white light; an optical lens provided on the optical path of the white light reflected from the color-discriminating mirror to converge the white light; And it may include an imaging device that acquires the cell image from white light collected by the optical lens.

The optical twizing device includes a condensing lens that focuses a laser beam output from the laser device as a parallel beam; a first reflector provided on the optical path of the laser beam focused by the condenser lens to reflect the laser beam toward the color-discriminating mirror; and a second reflector provided on the optical path of the laser beam that has passed through the color-discriminating mirror to reflect the laser beam toward the cell. The white light condensed by the objective lens may be reflected from the second reflector and incident on the color line mirror.

The method for detecting cell capacitance based on simultaneous optical twigging according to an embodiment of the present invention is to image the cell on the pair of electrodes and the substrate by the image acquisition device and adjust the position of the cell for optical twigging. Obtaining a possible cell image; The focused laser beam is irradiated using the laser device of the optical twizing device, and the irradiation position of the focused laser beam is adjusted with respect to the substrate using the cell image to control the position of the cell with respect to the pair of electrodes. adjusting; and analyzing the characteristics of the cell by measuring capacitance between the pair of electrodes using an analyzer of the cell analysis device.

In the step of adjusting the position of the cell, when the first cell array to be analyzed is selected from among a plurality of preset cell arrays, one or more first cells are selected from among the plurality of cells included in the cell image according to the first cell array. steps; Measuring a first coordinate corresponding to the position of the first cell in the cell image, and determining a target position for each of the first cells based on the first cell array and the first coordinate; and moving at least one of the substrate and the laser device according to the target position to adjust the position of the first cells to match the first cell arrangement.

Analyzing the characteristics of the cells includes measuring a first capacitance value by measuring impedance between the pair of electrodes while the cells are arranged in the first cell array on the substrate; calculating a second capacitance value representing the capacitance of a single cell by applying the first capacitance value to a correction function set for the first cell array; and analyzing the characteristics of the cell based on the second capacitance value. The characteristics of the cell may include at least one of the type of the cell, the size of the cell, the electric charge of the cell, and electrostatic interaction between cells.

According to an embodiment of the present invention, an in situ cell capacitance sensing device and method are provided for detecting capacitance for a single cell or an array of multiple cells based on simultaneous optical tweezing.

Meanwhile, the effects that can be obtained from the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below. You will be able to.

1 is a configuration diagram of a simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention.
Figure 2 is a conceptual diagram of a cell analysis device constituting a simultaneous optical twizing-based cell capacitance detection device according to an embodiment of the present invention.
Figure 3 is a cross-sectional view showing a substrate and a pair of electrodes constituting a simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention.
Figure 4 is a microscope image showing a cell arrangement obtained by optical trapping according to an embodiment of the present invention.
Figure 5 is an exemplary diagram for explaining a process of measuring capacitance related to a cell according to an embodiment of the present invention.
Figure 6 is a graph showing the results of measuring capacitance for various cell arrangements according to an embodiment of the present invention.
Figure 7 is a graph showing the ratio measurement results for single cell capacitance measured for various cell arrangements according to an embodiment of the present invention.

Other advantages and features of the present invention and methods for achieving them will become clear by referring to the embodiments described in detail below along with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below and may be implemented in various different forms. The present embodiments are merely provided to ensure that the disclosure of the present invention is complete and to provide common knowledge in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

Even if not defined, all terms (including technical or scientific terms) used herein have the same meaning as generally accepted by the general art in the prior art to which this invention belongs. Terms defined by general dictionaries may be interpreted as having the same meaning as they have in the related art and/or text of the present application, and should not be conceptualized or interpreted in an overly formal manner even if expressions are not clearly defined herein. won't

The configuration of the invention to clarify the solution to the problem to be solved by the present invention will be described in detail with reference to the accompanying drawings based on preferred embodiments of the present invention, and the same will be true in assigning reference numbers to the components in the drawings. Components are given the same reference numbers even if they are in different drawings, and it is stated in advance that components of other drawings can be cited when necessary when explaining the relevant drawings.

1 is a configuration diagram of a simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention. Referring to FIG. 1, the simultaneous optical tweezing-based cell capacitance sensing device 100 according to an embodiment of the present invention detects capacitance for a single cell or multiple cell arrays based on simultaneous optical tweezing. It is for in situ cell capacitance detection, and for this purpose, it may include an optical twirling device 110, an image acquisition device 120, and a cell analysis device 130.

Figure 2 is a conceptual diagram of a cell analysis device constituting a simultaneous optical twizing-based cell capacitance detection device according to an embodiment of the present invention. 1 and 2, the cell analysis device 130 is used to measure the capacitance of the area where the cell 10 is located and to analyze the characteristics of the cell 10 from the capacitance. It may include a substrate 131, a pair of electrodes 132 and 133, an analyzer 134, and a controller 135.

The substrate 131 may be prepared on which the cells 10 are placed. Cells 10 may include a single cell or multiple cells. In order to effectively deliver a focused laser beam for optical tweezing (optical tweezers) to the cells 10 and obtain cell images by imaging the area containing the cells 10, the substrate 131 is made of a light-transmitting material such as quartz. It can be provided as a transparent substrate made of material.

Figure 3 is a cross-sectional view showing a substrate and a pair of electrodes constituting a simultaneous optical twizing-based cell capacitance sensing device according to an embodiment of the present invention. Referring to FIGS. 1 to 3 , a pair of electrodes 132 and 133 may be arranged to be spaced apart from each other on both sides of the substrate 131 . A pair of electrodes 132 and 133 may be arranged to be spaced apart by a distance that allows single cells or multiple cells to be aligned. In order to ensure the accuracy of cell characteristic analysis through measurement of capacitance, the separation distance (G) between a pair of electrodes 132 and 133 may be designed to range from 10 ㎛ to 200 ㎛, for example.

A pair of electrodes 132 and 133 may be made of a conductive material such as metal (eg, Au, Cr, etc.). In an embodiment, each electrode 30 constituting the pair of electrodes 132 and 133 is a single electrode layer or top and bottom on the electrode area 32 excluding the cell area 40 where the cells of the substrate 131 are placed. It may be composed of a plurality of stacked electrode layers 34, 36, and 38. The thickness of the electrode 30 may be designed to be approximately 100 nm to 1 um.

The analyzer 134 can analyze the characteristics of the cell 10 by measuring the capacitance between a pair of electrodes 132 and 133. The analyzer 134 determines the type of cell 10, the size of the cell 10, the charge of the cell 10, and/or the electrostatic interaction between the cells based on the capacitance between the pair of electrodes 132 and 133. The characteristics of the cell 10, such as the like, can be analyzed.

The position of the cell 10 based on the pair of electrodes 132 and 133, the number of cells 10 located between the pair of electrodes 132 and 133, and the cell arrangement are determined by the measured capacitance and this. It affects the results of cell characterization analysis. Therefore, in order to accurately analyze cell characteristics and further analyze various cell characteristics through capacitance measurement for various cell arrangements, it is necessary to accurately control the arrangement of cells based on a pair of electrodes 132 and 133. .

In order to accurately control the arrangement of cells between a pair of electrodes 132 and 133, an optical twirling device 110 and an image acquisition device 120 may be provided. The image acquisition device 120 captures a cell 10 on a pair of electrodes 132 and 133 and the substrate 131 to produce a cell image that can be referenced for position adjustment (or cell array control) of the cell 10. It can be obtained.

The image acquisition device 120 may include a light source 121, a focusing lens 122, an objective lens 123, a dichroic mirror 124, one or more optical lenses 125, 126, and an imaging device 127. You can. The light source 121 is provided on the top of the substrate 131 to acquire cell images and can output white light toward the cells 10 on the substrate 131. A condenser lens 122 is provided on the optical path of the white light output from the light source 121 and can focus the white light toward the cells 10 on the substrate 131.

An objective lens 123 may be provided on the optical path of the white light passing through the substrate 131 to converge the white light. White light collected by the objective lens 123 may be reflected by a second reflector 115, which will be described later, and be incident on a dichroic mirror 124. The color-screening mirror 124 may be provided on the optical path of the white light collected by the objective lens 123 to reflect the white light.

The optical lenses 125 and 126 are provided on the optical path of the white light reflected from the color-discriminating mirror 124 and can focus the white light. The optical lenses 125 and 126 may be composed of, for example, convex lenses, concave lenses, etc. The imaging device 127 may acquire cell images from white light collected by the optical lenses 125 and 126. The imaging device 127 may be comprised of, for example, a CCD camera or a CMOS camera. Cell images acquired by the imaging device 127 may be output to the controller 135.

The optical tweezing device 110 radiates a focused laser beam to trap cells based on the optical tweezers principle, and adjusts the irradiation position of the focused laser beam 20 with respect to the substrate 131 to form a pair of electrodes 132, 133), the position of the cell 10 can be adjusted. To this end, the optical tweezing device 110 may include a laser device 111, one or more condensing lenses 112 and 113, a first reflector 114, and a second reflector 115.

The laser device 111 can irradiate a focused laser beam to trap cells using an optical tweezers method. The laser device 111 may output a parallel laser beam using a collimator. The condensing lenses 112 and 113 may focus the laser beam output from the laser device 111 as a parallel beam. The condensing lenses 112 and 113 may be composed of convex lenses, concave lenses, etc.

The first reflector 114 is provided on the optical path of the laser beam focused by the condensing lenses 112 and 113 and can reflect the laser beam toward the color-discriminating mirror 124. The second reflector 115 is provided on the optical path of the laser beam that has passed through the color-discriminating mirror 124 and can reflect the laser beam toward the cell 10. The laser beam (focused laser beam) reflected from the second reflector 115 may be incident on the substrate 131 through the objective lens 123 and transmitted to the cells 10 through the substrate 131.

The optical tweezing device 110 can adjust the position of the cell 10 with respect to the pair of electrodes 132 and 133 by adjusting the irradiation position of the focused laser beam with respect to the substrate 131. In an embodiment, the substrate 131 may be configured to be movable in a first horizontal direction and a second horizontal direction perpendicular to the first horizontal direction. That is, the optical twizing device 110 may include a driving device (not shown) such as a driving motor or a driving cylinder that moves the substrate 131 in the horizontal direction on the X-Y stage.

The optical twirling device 110 determines the position (first two-dimensional coordinates) of the cell 10 extracted from the cell image acquired by the image acquisition device 120 from the controller 135 and the focused laser beam 20. The position (second two-dimensional coordinates) is input, and the position of the cell 10 in the cell image is determined according to the differential position (two-dimensional coordinate deviation) between the position of the cell 10 and the position of the focused laser beam 20. The substrate 131 may be moved to match the position of the beam 20.

Accordingly, when the focused laser beam 20 is focused on the cell 10, a force to trap the cell 10 is applied due to the interaction between the optical energy of the focused laser beam 20 and the cell 10. Accordingly, the cell 10 is confined to the focused laser beam 20. In this state, when the substrate 131 is moved again, the cells 10 are restrained by the focused laser beam 20 and do not move, and the electrodes 132 and 133 on the substrate 131 are moved and the electrodes 132 and 133 are moved. ) can be adjusted.

At this time, the amount of movement of the substrate 131 in the . The current position of the cell 10 can be analyzed from the cell image by the controller 135, and the target position of the cell 10 can be determined according to the cell position selected (input) by the user or the cell arrangement.

In one embodiment, when the first cell array to be analyzed among a plurality of preset cell arrays is selected by the user, the optical twirling device 110 operates on one or more cells among a plurality of cells included in the cell image according to the first cell array. The first cell may be selected. At this time, the plurality of cell arrays may include, for example, a single cell array, a horizontal/vertical double cell array, a horizontal/vertical triple cell array, and a multiple matrix cell array (a structure in which two or more cells are arranged in the horizontal and vertical directions, respectively). there is.

The optical twirling device 110 executes a control command to the controller 135 to measure the first coordinate corresponding to the position of the first cell in the cell image, and determines the first cell based on the first cell array and the first coordinate. The target position can be determined for each. The optical tweezing device 110 may adjust the position of the first cells to match the first cell arrangement by moving the substrate 131 and/or the laser device 111 according to the determined target position. The optical tweezing device 110 may sequentially adjust the position of one or more first cells on the substrate 131 in a specified order according to the first cell arrangement so that the one or more first cells form the first cell arrangement. .

The analyzer 134 may measure the first capacitance value by measuring the impedance between a pair of electrodes 132 and 133 while the cells are arranged in a first cell array on the substrate 131. The analyzer 134 calculates a second capacitance value representing the capacitance of a single cell by applying the measured first capacitance value to the correction function set for the first cell array, and based on the calculated second capacitance value. You can analyze the characteristics of cells. At this time, the characteristics of the cell analyzed by the analyzer 134 may include the type of the cell, the size of the cell, the electric charge of the cell, and electrostatic interactions between cells.

According to the embodiment of the present invention as described above, the distance between the cell and the electrodes 132 and 133 can be adjusted according to the purpose by optical tweezers technology, and the distance between the cell and the electrodes 132 and 133, the pair of electrodes (132, 133) The characteristics of cells or interactions between cells can be accurately analyzed by measuring capacitance related to cells while controlling the number of cells, spacing between cells, and arrangement of cells. there is.

According to an embodiment of the present invention, the position of cells can be adjusted using a focused laser beam with optical forces of scattering and tilting forces without the need to directly contact the cells, and living cells trapped in optical tweezers can be prevented from physical damage. Various cell characteristics can be analyzed by adjusting cell position and/or cell arrangement while maintaining function.

The cell trapping technology according to embodiments of the present invention can be used to effectively monitor interactions between nearby cells, which are essential for the development and regulation of living cells. In addition, according to embodiments of the present invention, capacitance measurement data can be processed to analyze a single cell of interest, and cells can be selected based on the capacitance measured for multiple cells at a specific cell spacing and specific cell alignment. It can be used to investigate inter-electrostatic interactions.

Hereinafter, an experiment to verify the performance of the simultaneous optical twizing-based cell capacitance sensing device and method according to an embodiment of the present invention will be described. To measure the effective capacitance of optically trapped cells in a non-invasive, label-free and rapid manner, a simultaneous optical twigging-based cell capacitance sensing device as shown in Figure 1 was fabricated to measure the capacitance associated with the cells. . A quartz substrate was used as a substrate to support cells, and a linear electrode was used as an electrode for measuring capacitance.

Measurement conditions were carefully adjusted to ensure that there was no cell damage induced by the focused laser beam or electroporation. Capacitance values were calculated for various cell numbers and cell alignments (single, double, triple, and multiple cells in horizontal, vertical, or mixed alignment) and corrected to yield the capacitance value corresponding to a single cell. Capacitance values were measured for single cells, double, triple, and multi-arranged cells arranged vertically/horizontally, and the change in capacitance according to cell arrangement or separation distance between cells was analyzed.

Optical tweezing was performed to place cells between a pair of electrodes. Optical tweezers were utilized to capture and move single cells without contact using a highly focused beam. Real-time cell images and capacitance data were collected using a computer controller connected to a CCD camera and analyzer.

Figure 4 is a microscope image showing a cell arrangement obtained by optical trapping according to an embodiment of the present invention. Cell imaging can be utilized to ensure that capacitance data was acquired under the conditions of the cells of interest and the cell array of interest without the possibility of unwanted cell movement or disturbed cell alignment.

A system setup combining optical tweezers and an impedance analyzer is useful for measuring the capacitance of different numbers of cells (e.g. 1, 2, 3, 4, 6, and 10 cells) and provides accurate Optical tweezing allows cells to be positioned in a variety of ways, at specific distances (adjacent or spacing), and according to specific cell alignments (horizontal, vertical, or mixed arrangements). As shown in Figure 4, it was possible to observe single, double, triple, and multiple cells with various cell arrangements.

In Figure 4 and the drawings below, 'DC' is a double cell, 'DCve' is a vertically aligned double cell, 'DS' is a spaced (spaced) double cell, 'TC' is a triple cell, and 'TCve' is a vertical cell. Aligned triple cells, 'TS' is spaced triple cells, 'QC' is 4 cells with a 2X2 cell arrangement, '3X2' is 6 cells with a 3X2 cell arrangement, '2X3' is 6 cells with a 2X3 cell arrangement. Cells, 'Multiple' represents 10 cells with a 5X2 cell arrangement.

Figure 5 is an exemplary diagram for explaining a process of measuring capacitance related to a cell according to an embodiment of the present invention. Figure 6 is a graph showing the results of measuring capacitance for various cell arrangements according to an embodiment of the present invention. Figure 7 is a graph showing the ratio measurement results for single cell capacitance measured for various cell arrangements according to an embodiment of the present invention. The equations assigning the capacitance values (Cm) measured for various cell arrays and the capacitance values (C _B ) measured with only phosphate-buffered saline (Dulbecco's phosphate-buffered saline (DPBS)) solution without cell arrays to each cell array. For example, the capacitance value (Cx) associated with an individual cell can be calculated by applying the correction functions of Equations 1 to 6 below.

[Equation 1]

[Equation 2]

[Equation 3]

[Equation 4]

[Equation 5]

[Equation 6]

Equation 1 is the capacitance correction function for a single cell, Equation 2 is the capacitance correction function for double cells (DC) and spaced double cells (DS), and Equation 3 is the capacitance correction function for vertically aligned double cells (DCve). The capacitance correction function for, Equation 4 is the capacitance correction function for triple cells (TC) and spaced triple cells (TS), Equation 5 is the capacitance correction function for vertically aligned triple cells (TCve), math Equation 6 is the capacitance correction function for 2X2 multiple cells (QC). The correction function can be preset or calculated through data statistical analysis or regression analysis.

Capacitance values (Cx) corrected using a correction function can show significant differences between cell array groups. The capacitance value (Cx) obtained for a single cell (red line) showed the lowest value at all scanned frequencies. The capacitance values (Cx) obtained for the double cell (orange line) show a significant increase, and the capacitance values (Cx) obtained for the triple cell (blue line) also show a significant increase.

Looking at the capacitance ratio plot (see Figure 7), where the capacitance values obtained for individual cell arrays (Cx) are divided by the capacitance values for single cells (Cx), we see a ~1.6-fold increase for dual cells; , it can be seen that triple cells show a ~2.5-fold increase compared to single cell Cx values. An increase in capacitance value may indicate larger cell size or a stronger charge on the cell. Additionally, the capacitance values obtained for cell arrays can be utilized to analyze electrostatic interactions between cells.

The terminology used herein is for describing embodiments and is not intended to limit the invention. As used herein, singular forms also include plural forms, unless specifically stated otherwise in the context. Terms such as 'unit' and 'unit' used throughout this specification may refer to a unit that processes at least one function or operation, for example, a hardware component such as software, FPGA, or ASIC. '~ part', '~ part', etc. may be configured to reside in an addressable storage medium and may be configured to reproduce one or more processors.

The above detailed description is illustrative of the present invention. Additionally, the foregoing is intended to illustrate preferred embodiments of the present invention, and the present invention can be used in various other combinations, modifications, and environments. That is, changes or modifications can be made within the scope of the inventive concept disclosed in this specification, within the scope equivalent to the written disclosure, and/or within the scope of technology or knowledge in the art. The written examples illustrate the best state for implementing the technical idea of the present invention, and various changes required for specific application fields and uses of the present invention are also possible. Accordingly, the detailed description of the invention above is not intended to limit the invention to the disclosed embodiments. Additionally, the appended claims should be construed to include other embodiments as well.

10: cells
20: Focused laser beam
30: electrode
100: Cellular capacitance sensing device based on simultaneous optical twizing
110: Optical twizing device
111: Laser device
112, 113: condenser lens
114: first reflector
115: second reflector
120: Image acquisition device
121: light source
122: Focusing lens
123: Objective lens
124: Color-selected mirror
125, 126: Optical lens
127: imaging device
130: Cell analysis device
131: substrate
132, 133: electrode
134: Analyzer
135: controller

Claims (10)

A cell analysis device comprising a substrate on which cells are placed, a pair of electrodes spaced apart on both sides of the substrate, and an analyzer configured to analyze the characteristics of the cells by measuring capacitance between the pair of electrodes. ;
an optical twizing device including a laser device that irradiates a focused laser beam for trapping, and configured to adjust the position of the cell with respect to the pair of electrodes by adjusting an irradiation position of the focused laser beam with respect to the substrate; and
An image acquisition device configured to capture cells on the pair of electrodes and the substrate to acquire cell images that can be used as a reference for adjusting the position of the cells,
The optical tweezing device:
moving the substrate so that the position of the cell in the cell image matches the position of the focused laser beam according to the differential position between the position of the cell and the position of the focused laser beam; and
By moving the substrate in a state where the cells are not moved by binding the cells to the focused laser beam due to the interaction between the optical energy of the focused laser beam and the cells, the pair of electrodes on the substrate is moved to form cells of the cells. A cell capacitance sensing device based on simultaneous optical twigging that adjusts the position of each cell relative to the pair of electrodes to conform to the array. In claim 1,
The optical tweezing device:
When a first cell array to be analyzed is selected from among a plurality of preset cell arrays, selecting one or more first cells from among a plurality of cells included in the cell image according to the first cell array;
Measure a first coordinate corresponding to the position of the first cell in the cell image, and determine a target position for each first cell based on the first cell array and the first coordinate;
Simultaneous optical tweezing based cell capacitance sensing device configured to adjust the position of the first cell to match the first cell arrangement by moving at least one of the substrate and the laser device according to the target position. In claim 2,
The plurality of cell arrays may include a single cell array; horizontal double cell arrangement; vertical double cell array; horizontal triple cell arrangement; Vertical triple cell array; and a multi-matrix cell array in which two or more cells are arranged in each of the horizontal and vertical directions. A cell capacitance sensing device based on simultaneous optical tweezing including both. In claim 2,
The analyzer:
measuring a first capacitance value by measuring impedance between the pair of electrodes while the cells are arranged in the first cell array on the substrate;
Applying the first capacitance value to a correction function set for the first cell array to calculate a second capacitance value representing the capacitance of a single cell;
A simultaneous optical tweezing based cell capacitance sensing device configured to analyze characteristics of the cell based on the second capacitance value. In claim 1,
The image acquisition device:
a light source that outputs white light toward cells on the substrate to acquire images of the cells;
a focusing lens provided on the optical path of the white light output from the light source to focus the white light toward the cells on the substrate;
an objective lens provided on the optical path of the white light passing through the substrate to converge the white light;
a color-selective mirror provided on the optical path of the white light condensed by the objective lens to reflect the white light;
an optical lens provided on the optical path of the white light reflected from the color-discriminating mirror to converge the white light; and
Simultaneous optical tweezing-based cell capacitance sensing device comprising an imaging device that acquires an image of the cell from white light collected by the optical lens. In claim 5,
The optical tweezing device:
a condensing lens that focuses the laser beam output as parallel light from the laser device;
a first reflector provided on the optical path of the laser beam focused by the condenser lens to reflect the laser beam toward the color-discriminating mirror; and
A second reflector provided on the optical path of the laser beam passing through the color-discriminating mirror to reflect the laser beam toward the cell,
The white light collected by the objective lens is reflected from the second reflector and is incident on the dichroic mirror. A simultaneous optical twizing-based cell capacitance sensing method performed by the simultaneous optical twizing-based cell capacitance sensing device of any one of claims 1 to 6, comprising:
acquiring, by the image acquisition device, images of cells on the pair of electrodes and the substrate to obtain cell images that can be referenced for position adjustment for optical twirling of the cells;
The focused laser beam is irradiated using the laser device of the optical twizing device, and the irradiation position of the focused laser beam is adjusted with respect to the substrate using the cell image to control the position of the cell with respect to the pair of electrodes. adjusting; and
Comprising: analyzing the characteristics of the cell by measuring the capacitance between the pair of electrodes using an analyzer of the cell analysis device,
The steps for adjusting the position of the cell are:
moving the substrate so that the position of the cell in the cell image matches the position of the focused laser beam according to the differential position between the position of the cell and the position of the focused laser beam; and
By moving the substrate in a state where the cells are not moved by binding the cells to the focused laser beam due to the interaction between the optical energy of the focused laser beam and the cells, the pair of electrodes on the substrate is moved to form cells of the cells. A method for sensing cell capacitance based on simultaneous optical tweezing, comprising adjusting the position of the cell relative to the pair of electrodes to conform to the array. In claim 7,
The steps for adjusting the position of the cell are:
When a first cell array to be analyzed is selected from among a plurality of preset cell arrays, selecting one or more first cells from among a plurality of cells included in the cell image according to the first cell array;
Measuring a first coordinate corresponding to the location of the first cell in the cell image, and determining a target location for each of the first cells based on the first cell array and the first coordinate; and
Simultaneous optical tweezing-based cell capacitance sensing method comprising adjusting the position of the first cell to match the first cell arrangement by moving at least one of the substrate and the laser device according to the target position. In claim 8,
The steps for analyzing the characteristics of the cells are:
measuring a first capacitance value by measuring impedance between the pair of electrodes while the cells are arranged in the first cell array on the substrate;
calculating a second capacitance value representing the capacitance of a single cell by applying the first capacitance value to a correction function set for the first cell array; and
Analyzing the characteristics of the cell based on the second capacitance value. In claim 9,
A method for detecting cell capacitance based on simultaneous optical tweezing, wherein the characteristics of the cell include at least one of the type of the cell, the size of the cell, the charge of the cell, and electrostatic interactions between cells.

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