A real-time cell monitoring system

Abstract

The present application provides a real-time cell monitoring system which can improve the accuracy and convenience of real-time cell analysis by simultaneously measuring various electrical signals and oxygen consumption rate for cells in a non-destructive manner.

Description

Technical Field

[0001] This invention relates to a real-time cell monitoring system, and more specifically, to a real-time cell monitoring system that simultaneously measures multiple electrical signals and oxygen consumption rates of cells using a non-destructive method, thereby improving the accuracy and convenience of real-time cell analysis. Background Technology

[0002] In modern biological and medical research, cell analysis has become an indispensable element. In particular, the accurate analysis of cell state—a crucial biological process in which cells transform into mature forms with specific functions—plays a vital role in various research and experiments, including disease diagnosis, treatment development, and drug efficacy evaluation.

[0003] Previous cell analyses utilized methods such as microscopic observation and biochemical analysis, while cell differentiation assessment employed methods such as gene expression analysis, protein expression profiling, immunofluorescence, and flow cytometry. However, these methods not only struggle to monitor real-time cell dynamics but also suffer from limitations in terms of cell contamination, accuracy, cost, and time.

[0004] Recently, we have been researching various non-destructive real-time cell analysis techniques, such as electrical cell-substrate impedance sensing (ECIS), local field potential measurement, and oxygen consumption rate (OCR) measurement. However, while these techniques can analyze the functional state and changes of cells in real time, most of them are limited to the level of individual measurement of specific data.

[0005] Therefore, there is a need to develop a new system that can more accurately and in real time monitor and analyze cell states such as cell metabolism, cell culture, cell differentiation, and drug screening. Summary of the Invention

[0006] The problem that the invention aims to solve The technical problem to be solved by the present invention is to provide a real-time cell monitoring system that simultaneously measures multiple electrical signals and oxygen consumption rate of cells in a non-destructive manner, thereby improving the accuracy and convenience of real-time cell analysis.

[0007] The technical problems of this invention are not limited to those mentioned above. Those skilled in the art can clearly understand other technical problems not mentioned through the following description.

[0008] means for solving problems To address the aforementioned technical problems, a real-time cell monitoring system according to an embodiment of the present invention for simultaneously measuring cell impedance and oxygen consumption rate includes: a cell electrical signal measurement module comprising an impedance measurement unit and a controller; a cell chip fixed to the cell electrical signal measurement module and comprising: a hole having a receiving portion open at the upper end to accommodate cells and culture medium; a substrate disposed below the hole and having a common electrode and a working electrode formed thereon; a guide module fixed to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip and having an opening to expose the upper end of the hole; a plate module fixed to the upper part of the guide module and including a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and an oxygen consumption rate measurement module fixed to the upper part of the plate module and including a light-emitting unit for emitting light to the oxygen-responsive substance and a sensor unit for receiving optical signals generated from the oxygen-responsive substance.

[0009] The impedance measuring unit can be electrically connected to the common electrode and the working electrode, and can measure the impedance between the common electrode and the working electrode by applying an AC signal to the common electrode.

[0010] The probe can be located inside the receiving portion of the hole.

[0011] The oxygen consumption rate measuring module may also include a driving component that moves the plate module up and down, and the distance between the probe and the bottom of the receiving part of the hole may change according to the up and down movement of the plate module.

[0012] The system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, the processor being able to analyze the state of the cells using impedance data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

[0013] To achieve the aforementioned technical problem, a real-time cell monitoring system according to another embodiment of the present invention for simultaneously measuring local field potential and oxygen consumption rate includes: a cell electrical signal measurement module, which includes a local field potential measurement unit and a controller; a cell chip, which is fixed to the cell electrical signal measurement module and includes: a hole having a receiving portion open at the upper end to accommodate cells and culture medium; a substrate disposed at the lower part of the hole and having a common electrode and a working electrode formed thereon; a guide module, which is fixed to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip and has an opening to expose the upper end of the hole; a plate module, which is fixed to the upper part of the guide module and includes a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and an oxygen consumption rate measurement module, which is fixed to the upper part of the plate module and includes a light-emitting unit for emitting light to the oxygen-responsive substance and a sensor unit for receiving optical signals generated from the oxygen-responsive substance.

[0014] The local field potential measuring unit can be electrically connected to the common electrode and the working electrode, and can measure the local field potential between the common electrode and the working electrode.

[0015] The probe can be located inside the receiving portion of the hole.

[0016] The oxygen consumption rate measuring module may also include a driving component that moves the plate module up and down, and the distance between the probe and the bottom of the receiving part of the hole may change according to the up and down movement of the plate module.

[0017] The system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, the processor being able to analyze the state of the cell by utilizing local field potential data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

[0018] To achieve the aforementioned technical problem, a real-time cell monitoring system according to another embodiment of the present invention for simultaneously measuring cell impedance, local field potential, and oxygen consumption rate includes: a cell electrical signal measurement module, which includes an impedance measurement unit, a local field potential measurement unit, and a controller; a cell chip, which is fixed to the cell electrical signal measurement module and includes: a hole having a receiving portion open at the upper end to accommodate cells and culture medium; and a substrate disposed at the lower part of the hole and having a common electrode and a working electrode formed thereon; a guide module, which is fixed to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip and has an opening to expose the upper end of the hole; a plate module, which is fixed to the upper part of the guide module and includes a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and an oxygen consumption rate measurement module, which is fixed to the upper part of the plate module and includes a light-emitting unit for emitting light to the oxygen-responsive substance and a sensor unit for receiving optical signals generated from the oxygen-responsive substance.

[0019] The impedance measuring unit and the local field potential measuring unit can be electrically connected to the common electrode and the working electrode. The impedance measuring unit can measure the impedance between the common electrode and the working electrode by applying an AC signal to the common electrode. The local field potential measuring unit can measure the local field potential between the common electrode and the working electrode.

[0020] The probe can be located inside the receiving portion of the hole.

[0021] The oxygen consumption rate measuring module may also include a driving component that moves the plate module up and down, and the distance between the probe and the bottom of the receiving part of the hole may change according to the up and down movement of the plate module.

[0022] The system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, the processor being able to analyze the state of the cell by utilizing impedance data and local field potential data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

[0023] Invention Effects According to the present invention as described above, it is possible to simultaneously monitor cell impedance and oxygen consumption rate, local field potential and oxygen consumption rate, or cell impedance, local field potential and oxygen consumption rate in real time using a non-destructive method.

[0024] In addition, it has the ability to more accurately analyze cell states such as cell metabolism, cell culture, cell differentiation, and drug screening by utilizing simultaneously measured cell impedance, local field potential, or oxygen consumption rate data.

[0025] The effects of the present invention are not limited to those mentioned above, and those skilled in the art can clearly understand other effects not mentioned from the following description. Attached Figure Description

[0026] Figure 1 This is a schematic perspective view of a real-time cell monitoring system according to an embodiment of the present invention.

[0027] Figure 2 It is shown in a general way. Figure 1 An exploded 3D diagram of the components of a real-time cell monitoring system.

[0028] Figure 3 (a) to Figure 3 (c) is a diagram illustrating a cell electrical signal measurement module constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0029] Figure 4 This is a diagram illustrating one embodiment of a cell chip constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0030] Figure 5 It shows the composition Figure 4 A diagram of one embodiment of a substrate for a cell chip.

[0031] Figure 6 (a) and Figure 6 (b) is a diagram illustrating an embodiment of a boot module constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0032] Figure 7 This is a diagram illustrating one embodiment of a plate module and an oxygen consumption rate measurement module constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0033] Figure 8 This is a graph showing the impedance results measured using a real-time cell monitoring system according to an embodiment of the present invention.

[0034] Figure 9 This is a graph showing the results of oxygen consumption rate measured using a real-time cell monitoring system according to an embodiment of the present invention.

[0035] Figure 10 This is a diagram illustrating a cell electrical signal measurement module that constitutes a real-time cell monitoring system according to another embodiment of the present invention.

[0036] Figure 11 This is a diagram illustrating a cell electrical signal measurement module constituting a real-time cell monitoring system according to yet another embodiment of the present invention. Detailed Implementation

[0037] The advantages, features, and methods of implementing the present invention will become apparent from the accompanying drawings and the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various different forms. These embodiments are provided only to complete the disclosure of the invention and to fully inform those skilled in the art of the scope of the invention, which is defined solely by the scope of the claims.

[0038] The terminology used in this specification will be briefly explained, and then the invention will be described in detail.

[0039] Regarding the terminology used in this invention, widely used general terms have been selected to the extent possible while considering the functionality of the invention. However, this may vary depending on the intent of those skilled in the art, precedents, or the emergence of new technologies. Furthermore, in certain cases, the applicant may arbitrarily choose and use terms; in such cases, their meanings will be detailed in the corresponding sections of the specification. Therefore, the terminology used in this invention should not be defined based on its simple name, but rather on its meaning and the overall content of the invention.

[0040] Throughout this specification, when a section is referred to as "including" a component, unless specifically stated otherwise, it does not exclude other components, but rather indicates that other components may also be included. Furthermore, the terms "section," "module," and "unit" as used in this specification refer to a unit that performs at least one function or operation, which can be implemented by software, hardware components such as FPGAs or ASICs, or a combination of software and hardware. However, the terms "section," "module," and "unit" are not limited to software or hardware. "Sections," "modules," and "units" can be configured to reside in addressable storage media and can be configured to execute on one or more processors. Therefore, as an example, the terms "section," "module," and "unit" include components such as software components, object-oriented software components, class components, and task components, as well as procedures, functions, attributes, programs, subroutines, program code segments, drivers, firmware, microcode, circuits, data, databases, data structures, tables, arrays, and variables.

[0041] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings to enable those skilled in the art to readily implement the invention. Furthermore, in the drawings, parts unrelated to the description will be omitted for clarity.

[0042] Terms containing ordinal numbers, such as "first" and "second," can be used to describe various constituent elements, but the constituent elements are not limited by these terms. These terms are only used to distinguish one constituent element from other constituent elements. For example, without departing from the scope of the invention, a first constituent element can be named a second constituent element, and similarly, a second constituent element can be named a first constituent element. The term "and / or" includes a combination of multiple related items or any one of multiple related items.

[0043] Figure 1 This is a schematic perspective view of a real-time cell monitoring system according to an embodiment of the present invention. Figure 2 It is shown in a general way. Figure 1 An exploded 3D diagram of the components of a real-time cell monitoring system. Figure 3 (a) to Figure 3 (c) is a diagram illustrating a cell electrophysiological signal measurement module constituting a real-time cell monitoring system according to an embodiment of the present invention. Figure 4 This is a diagram illustrating one embodiment of a cell chip constituting a real-time cell monitoring system according to an embodiment of the present invention. Figure 5 It shows the composition Figure 4 A diagram of one embodiment of a cell chip substrate. Figure 6 (a) and Figure 6 Figure (b) is a diagram illustrating one embodiment of a boot module constituting a real-time cell monitoring system according to an embodiment of the present invention. Figure 7 This is a diagram illustrating one embodiment of a plate module and an oxygen consumption rate measurement module constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0044] Reference Figure 1 and Figure 2 According to an embodiment of the present invention, a real-time cell monitoring system (1) (hereinafter also referred to as the "system") includes a cell electrical signal measurement module (100), a cell chip (200), a guide module (300), a plate module (400), and an oxygen consumption rate measurement module (500). The system (1) according to an embodiment of the present invention is capable of simultaneously measuring cell impedance (hereinafter also referred to as "impedance" or "impedance data") and oxygen consumption rate. Specifically, cell impedance can be measured at the lower part of the cell chip (200), and oxygen consumption rate can be measured at the upper part of the cell chip (200).

[0045] In addition, the system (1) may also include a processor (not shown) electrically connected to the cell electrical signal measurement module (100) and the oxygen consumption rate measurement module (500). The processor can analyze the state of the cells by using impedance data measured by the cell electrical signal measurement module (100) and oxygen consumption rate data measured by the oxygen consumption rate measurement module (500).

[0046] At this point, the system (1) can be implemented by the processor and multiple objects, or by a single object. In the case of implementation by multiple objects, a series of processes for analyzing cell states can be executed separately, and various data can be sent and received via wired or wireless communication networks.

[0047] Reference Figure 3 The cell electrical signal measurement module (100) includes an impedance measurement unit (110) and a controller (120). The cell electrical signal measurement module (100) performs the following functions: applying an AC signal to the cell chip (200) to measure the impedance generated during cell culture or during the treatment of cultured cells, and generating impedance data. Specifically, when an AC signal is applied to the wells (210) of the cell chip (200) during cell culture, as the number of cells gradually increases with cell culture, the area of ​​the common electrode (221) and working electrode (222) covering the lower part of the well (210) increases, which physically impedes the current, thereby increasing the impedance. Conversely, when an AC signal is applied during the treatment of cells cultured in the wells (210) of the cell chip (200) with drugs or the like, as the number of cells gradually decreases with the treatment, the area of ​​the common electrode (221) and working electrode (222) covering the lower part of the well (210) decreases, thereby decreasing the impedance.

[0048] Reference Figure 3 In one embodiment of the invention, the impedance measurement unit (110) includes a signal generator (111), a digital-to-analog converter (112), and an analog-to-digital converter (113).

[0049] The signal generator (111) can generate AC signals controlled by the controller (120) described later, which can be configured together with the controller (120) or provided as a separate power supply and associated with the controller (120).

[0050] The digital-to-analog converter (112) (DAC) is electrically connected to a common electrode (221) corresponding to each cell chip (200) and can convert an AC signal generated from the signal generator (111) into an analog input signal, which is then applied to the common electrode (221) corresponding to each cell chip (200).

[0051] An analog-to-digital converter (ADC) (113) is electrically connected to a working electrode (222) corresponding to each cell chip (200). It can receive an analog response signal generated between a common electrode (221) and a working electrode (222), convert the analog response signal into a digital signal, and then send the digital signal to a controller (120). Alternatively, the digital signal converted in the ADC (113) can be sent directly to the controller (120) or sent to the controller (120) at the back end via an additional component.

[0052] Reference Figure 3 In another embodiment of the invention, as shown in (b), the impedance measuring unit (110) may further include: a first signal amplifier (114a) (AMP) that amplifies the analog input signal at the back end of the digital-to-analog converter (112); and a second signal amplifier (114b) that amplifies the analog response signal at the front end of the analog-to-digital converter (113). In this case, there may be multiple first signal amplifiers (114a) or second signal amplifiers (114b).

[0053] Reference Figure 3 In another embodiment of the invention, the impedance measurement unit (110) at the front end of the second signal amplifier (114b) includes multiple input channels and one output channel, and may also include a multiplexer (115) that connects any one of the multiple input channels to the output channel. In one embodiment of the invention, when the cell chip (200) is composed of multiple holes (210), a common electrode (221) and a working electrode (222) are disposed at the lower part of the receiving portion (212) of each hole (210). The multiplexer (115) can receive all the different analog response signals generated between the common electrode 221 and the working electrode (222) corresponding to each hole (210) and selectively output them.

[0054] The controller (120) can control the overall operation of the impedance measurement unit (110), specifically, it can control the frequency of the AC signal applied to the cell chip (200) through the impedance measurement unit (110) and the impedance measurement time.

[0055] In addition, the controller (120) can receive impedance information measured by the impedance measuring unit (110) to generate impedance data, and send the generated impedance data to the processor (not shown) described later.

[0056] Specifically, the controller (120) can generate any one of the following impedance data: resistive data, capacitive data, amplitude data, phase data, and combinations thereof. More specifically, the controller (120) can receive impedance information from the impedance measurement unit (110) in the form of a sum of a resistance value as the real part and an reactance value as the imaginary part, generate resistive data using the resistance value as the real part, generate capacitive data using the reactance value as the imaginary part, and generate amplitude data using both the resistance value as the real part and the reactance value as the imaginary part. Furthermore, the controller (120) can of course generate impedance data for cells and impedance data for culture medium, respectively.

[0057] Furthermore, the controller (120) can generate different data based on the control operation for the impedance measurement unit (110).

[0058] As an example, the controller (120) can control the frequency of the AC signal to a single value during the impedance measurement period of the impedance measurement unit (110), at which time the controller (120) can generate impedance data over time.

[0059] As another example, the controller (120) can control the frequency of the AC signal to multiple different values ​​between the minimum frequency and the maximum frequency during the impedance measurement period of the impedance measurement unit (110). At this time, the controller (120) can generate impedance data by time and / or by frequency.

[0060] As another example, the controller (120) can control the frequency of the AC signal to multiple different values ​​between the minimum and maximum frequencies during the impedance measurement by the impedance measurement unit (110), and simultaneously control the measurement time according to the frequency of the AC signal. At this time, the controller (120) can generate impedance data by time and / or by frequency. Thus, the more frequencies of the AC signal applied to the cell chip (200) and the lower the frequency values, the longer the measurement time is required, and the accuracy of the impedance data generated by this process can be improved.

[0061] As described above, the system (1) of the present invention can control the operation of the impedance measurement unit (110) in various ways according to the purpose of cell impedance measurement, and the controller (120) can generate different data according to the control operation, thus significantly improving the convenience of cell analysis.

[0062] Reference Figure 4 and Figure 5 The cell chip (200) includes: wells (210) for culturing or processing cultured cells; and a substrate (220) having a common electrode (221) and a working electrode (222). As an example, the wells (210) can be disposed on the substrate (220) by thermoforming at 60 to 70°C and 30 to 30 psi on a substrate (120) covered with double-sided adhesive tape of polyethylene, silicone, polyurethane, or polyester. However, this is not a limitation; the wells (210) can be fixed to the substrate (220) by various methods.

[0063] The pores (210) are for culturing cells or processing cultured cells, and are preferably made of a transparent biocompatible material. The biocompatible material can be a transparent plastic such as PDMS, PMMA, PET, or PC, but is not limited to these, and can be made of transparent glass or the like.

[0064] The pore (210) has a receiving portion (212), which is formed with an open top due to the partition (211), thereby being able to contain cells and culture medium. The cross-section of the receiving portion (212) can be formed in various shapes such as circular and square. In addition, the pore (210) can be composed of a single receiving portion (212), which forms a space inside through the partition (211) arranged in the vertical direction. Through the partition (211), multiple receiving portions (212) can be arranged adjacently or spaced apart in rows (horizontal direction) and columns (vertical direction). At this time, the number of receiving portions (212) can be set in various ways, such as 4, 6, 7, 12, 16, 24, 48, 96, 128, etc.

[0065] The substrate (220) serves as a support hole (210) and also receives AC signals from the cell electrical signal measurement module (100). The substrate (220) is a non-conductive substrate, and is preferably made of transparent materials such as glass or plastic for optical monitoring of cell culture or cell processing status.

[0066] A plurality of electrodes are formed on the surface of the substrate (220), which are divided into a common electrode (221) and a working electrode (222) spaced apart from the common electrode (221). A transmission line (223) and a terminal pad (224) may also be formed together with the common electrode (221) and the working electrode (222). On the other hand, in another embodiment of the present invention, a stimulation electrode (not shown) may also be formed on the surface of the substrate (220) at a position spaced apart from the common electrode (221) and the working electrode (222).

[0067] The common electrode (221) may include a main body and a recessed portion, the recessed portion being formed by recessing a portion of the lateral side of the main body inward. Preferably, the recessed portion is disposed at the lower part of the receiving portion (212) of the hole (210), and multiple recessed portions may be arranged at intervals on the lateral side of the main body. Furthermore, preferably, the recessed portion is a semi-circular arc shape with a certain curvature, but is not limited thereto; it may also be a semi-rectangular shape or a triangular shape recessed inward.

[0068] The working electrode (222) can be disposed within the recess formed in the common electrode (221), and can be formed into various shapes depending on the shape of the recess. As an example, when the recess is a semi-circular arc shape with a certain curvature, the working electrode (222) can be formed into a corresponding disk shape.

[0069] The working electrode (222) and the common electrode (221) are spaced apart, thereby enabling impedance to be generated based on the AC signal applied from the impedance measurement module (100) to the common electrode (221). More specifically, the working electrode (222) is spaced apart from the common electrode (221) at the lower part of the accommodating portion (212) of the well (210), thereby enabling the non-destructive collection and analysis of real-time impedance changes caused by cell culture or cell treatment performed in the accommodating portion (212) of the well (210).

[0070] The transmission line (223) connects the common electrode (221) and the working electrode (222) to the terminal pad (224). The terminal pad (224) is electrically connected to the impedance measuring unit (110) to receive AC signals, and AC signals can be applied to the common electrode (221) through the transmission line (223).

[0071] As in one embodiment of the present invention, when a common electrode (221) is formed with a plurality of recesses and a plurality of working electrodes (222) (see [reference]). Figure 5The transmission lines (223) connected to each working electrode (222) can be formed with different widths to account for the coupling and parasitic capacitance generated by adjacent transmission lines (223) and connected to the terminal pad (224). At the same time, they can also be formed with different intervals with adjacent transmission lines (223) depending on their distance from the terminal pad (224). More specifically, preferably, the transmission lines (223) connected to the working electrodes (222) farther away from the terminal pad (224) are formed with a wider width, and the transmission lines (223) connected to the working electrodes (222) closer to the terminal pad (224) are formed with a narrower width. Furthermore, preferably, the transmission line (223) connected to the working electrode (222) farther from the terminal pad (224) has a wider spacing from its adjacent transmission line (223) compared to the transmission line (223) connected to the working electrode (222) closer to the terminal pad (224). This minimizes well variation and improves the reliability of cell analysis. In this case, the width and length of the transmission line (223) can be determined based on the theoretically calculated values ​​of the sheet resistance shown in Formula 1.

[0072] [Mathematical Expression 1]

[0073] Among them, R sheet ε represents the sheet resistance of the transmission line (223), l represents the resistivity of the material of the transmission line (223), w represents the length of the current flowing between the working electrode (222) and the terminal pad (224), and h represents the thickness of the transmission line (223).

[0074] The common electrode (221), working electrode (222), transmission line (223) and terminal pad (224) can be formed as transmissive, reflective or semi-transmissive, and can be formed of biocompatible conductive material.

[0075] As an example, the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) can be formed by depositing a transparent metal oxide composed of indium tin oxide (ITO), indium zinc oxide (IZO), zinc oxide (ZnO), or indium thin zinc oxide (ITZO) onto a substrate (220) and then patterning it, or by depositing the transparent metal oxide together with photoresist and then etching it, or by laser patterning. In this case, the hole (210), the substrate (220), and the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) formed on the substrate (220) are all formed of transparent material, which has the effect of facilitating optical monitoring of cell culture or cell processing status.

[0076] On the other hand, when the common electrode 221 and the working electrode (222) are formed of transparent materials, any one of the following nanomaterials can be vapor-deposited onto their surfaces: gold nanoparticles (AuNPs), quantum dots (QDs), reduced graphene oxide (rGO), poly(3,4-ethylenedioxythiophene)-polystyrene sulfonate (PEDOT:PSS), polyaniline (PANI), carbon nanotubes (CNTs), and multi-walled carbon nanotubes (MWCNTs). In this case, the nanomaterial is preferably a biocompatible conductive material, but is not limited to the nanomaterials shown above, as long as it is a biocompatible conductive material. As the interfacial impedance of the common electrode or working electrode decreases through the vapor deposition of the nanomaterial, the sensitivity and accuracy of impedance measurement can be further improved.

[0077] As another example, the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) can be formed on the substrate (220) by printing metal nanowires made of biocompatible materials such as chromium (Cr), gold (Au), and palladium (Pd). In this case, the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) can improve productivity and economic efficiency compared to the case where they are all formed of transparent materials.

[0078] Reference Figure 2 and Figure 6 The guide module (300) is fastened to the cell electrical signal measurement module (100) to play the role of the configuration position of the guide plate module (400). Specifically, it is preferable to fasten the guide module (300) to the cell electrical signal measurement module (100) to cover the surrounding area of ​​the cell chip (200).

[0079] When the guide module (300) is fastened to the cell electrophysiology module (100), an opening (310) is formed so that the upper end of the hole (210) of the cell chip (200) is exposed. In addition, in order to ensure stability when fastened to the cell electrophysiology module (100), the guide module (300) may have a fastening protrusion (320) formed on the side of the lower surface. At this time, the cell electrophysiology module (100) may have a fastening groove (130) for the fastening protrusion (320) of the guide module (300) to be inserted, and may also have a fixing member (140) for fixing the guide module (300).

[0080] Furthermore, the guide module (300) can be divided into an upper frame with an opening (310) and a lower frame with a fastening protrusion (320). In this case, the lower frame can be made wider than the upper frame, thereby forming a space between the ends of the upper frame and the ends of the lower frame, to which the plate module (400) described later can be fastened.

[0081] Reference Figure 2 and Figure 7 The plate module (400) is secured to the upper part of the guide module (300), and the oxygen consumption rate measurement module (500) is secured to the upper part of the plate module (400). In one embodiment of the invention, the plate module (400) and the oxygen consumption rate measurement module (500) can measure the oxygen concentration of the culture medium contained in the wells (210) of the cell chip (200) over time at the upper part of the cell chip (200), and then use the measured oxygen concentration and the following mathematical formula 1 to determine the oxygen consumption rate (OCR).

[0082] [Mathematical Expression 2]

[0083] Where d[O2] / dt represents the rate of change of oxygen concentration per unit time, and V represents the volume of the culture medium.

[0084] The board module (400) includes a probe (410) and a frame (420).

[0085] The probe (410) may be a rod-shaped part extending along the depth direction of the receiving portion (212) of the hole (210) and having a certain length. In this case, the probe (410) may be formed inside the receiving portion (212) of the hole (210). Specifically, the probe (410) is formed in the lower region of the frame (420) corresponding to the opening (310) of the guide module (300), and preferably is formed in the center of the inside of the receiving portion (212) of the hole (210), and more preferably is formed at a certain distance from the bottom of the receiving portion of the hole (210).

[0086] In addition, to determine oxygen concentration, an oxygen-responsive material can be attached to the lower part of the probe (410). In this case, the oxygen-responsive material can be any of the following: metal complexes, organic fluorophores, optoelectronic materials, nanomaterials, optical fiber-based oxygen-sensitive materials, silica gel, and polymer matrix. However, it is not limited to these; any substance that generates or changes a signal in response to oxygen concentration can be used as an oxygen-responsive material.

[0087] In addition, the probes (410) can be formed in the same number as the receiving portions (212). In the case where, as in one embodiment of the present invention, 16 receiving portions are formed on one side and 16 on the other side, for a total of 32, the probes (410) can also be formed in total of 32.

[0088] The frame (420) can be divided into a main frame (421) and a support frame (422), which can be integrated or separate. A portion of the lower inner side of the main frame (421) is fastened to the upper part of the guide module (300), and the support frame (422) is fastened to the side of the guide module (300). Thus, the plate module (400) can be stably supported on the upper part of the guide module (300).

[0089] In addition, the frame (420) can move up and down in the vertical direction under external pressure. For this purpose, the support frame (422) of one embodiment of the present invention can be made of a telescopic or elastic member. However, it is not limited to this. The frame (420) can form a separate flexible member in the main frame (421), so that the frame (420) can move up and down in the vertical direction under external pressure.

[0090] At this time, the probe (410) moves up and down in the vertical direction inside the receiving part (212) of the hole (210) as the frame (420) moves up and down, and the distance between the probe (410) and the bottom surface of the receiving part (212) of the hole (210) can change. Preferably, even if the probe (410) moves downward in the receiving part (212) of the hole (210), the probe (410) does not contact the bottom of the receiving part (212) of the hole (210) and remains at a certain distance.

[0091] In addition, the plate module (400) may also include an optical lens (430). The optical lens (430) is disposed inside the main frame (421), and more specifically, may be disposed in the region corresponding to the position of the light-emitting part (510) and the sensor part (520) of the probe 410 and the oxygen consumption rate measuring module (500) described later.

[0092] The optical lens (430) can focus the light emitted from the light-emitting part (510) of the oxygen consumption rate measuring module (500) onto the oxygen-responsive material of the probe (410), thereby more accurately measuring the optical signal that changes with oxygen concentration, and can control the path of the optical signal generated by the oxygen-responsive material to effectively concentrate it onto the sensor part (520), thereby improving sensitivity.

[0093] In addition, the board module (400) may also include a temperature sensor (not shown), an atmospheric pressure sensor (not shown), and / or a humidity sensor (not shown). The temperature sensor, the atmospheric pressure sensor, and the humidity sensor may be configured adjacent to the probe (410) for measuring the temperature, atmospheric pressure, and / or humidity inside the housing (212) of the hole (210), and the accuracy of the oxygen consumption rate (OCR) can be further improved by utilizing the measured temperature, atmospheric pressure, and / or humidity values.

[0094] The oxygen consumption rate measuring module (500) includes a light-emitting part (510) and a sensor part (520). In one embodiment of the present invention, the light-emitting part (510) and the sensor part (520) are disposed on the upper part of the oxygen consumption rate measuring module (500), but are not limited thereto. They can be disposed on the lower part and configured to contact the upper end of the plate module (400).

[0095] The light-emitting part (510) emits light to the oxygen-responsive material attached to the lower part of the probe (410). It can be a light-emitting diode (LED), but is not limited to it. Various light-emitting elements can be used.

[0096] The sensor unit (520) receives an optical signal generated from an oxygen-responsive material attached to the underside of the probe (410). It can be an optical sensor, but is not limited to it. Various elements capable of sensing light can be used.

[0097] In addition, the oxygen consumption rate measuring module (500) may also include a drive member (not shown) that moves the plate module (400) up and down. The plate module (400) can move up and down in the vertical direction by the action of the drive member, thereby changing the distance between the probe (410) and the bottom of the receiving part (212) of the hole (210).

[0098] At this time, the driving component may include an actuator and a driving pin connected to the actuator. The actuator may be configured on one side and / or the other side of the oxygen consumption rate measuring module (500), and the driving pin may be configured to be connected to the support frame (422) of the plate module (400). However, it is not limited to this. The driving component may be composed of various structures capable of moving the plate module (400) up and down, and may be configured in a variety of ways.

[0099] Furthermore, the oxygen consumption rate measurement module (500) may also include a control unit (not shown) and a communication unit (not shown). The control unit can control the light intensity of the light-emitting unit (510) and can measure (calculate) the oxygen concentration and oxygen consumption rate using the optical signal received by the sensor unit (520). The communication unit can transmit the oxygen concentration and oxygen consumption rate data measured (calculated) by the control unit to the processor via wired or wireless means.

[0100] Figure 8 This is a graph showing the impedance results measured using a real-time cell monitoring system according to an embodiment of the present invention. Figure 9 This is a graph showing the results of oxygen consumption rate measured using a real-time cell monitoring system according to an embodiment of the present invention.

[0101] Reference Figure 8 and Figure 9 According to an embodiment of the present invention, the system (1) can simultaneously measure cell impedance in the cell electrical signal measurement module (100) and oxygen consumption rate in the oxygen consumption rate measurement module (500) under the same cell assay conditions, thereby generating impedance data and oxygen consumption rate result charts over time, respectively. Furthermore, the system (1) according to an embodiment of the present invention can overlay the measured impedance data over time and the oxygen consumption rate result chart over time to generate a single chart. Therefore, the present invention has the effect of more accurately analyzing cell states such as cell metabolism, cell culture, cell differentiation, and drug screening.

[0102] Figure 10 This is a diagram illustrating a cell electrical signal measurement module that constitutes a real-time cell monitoring system according to another embodiment of the present invention.

[0103] Reference Figure 10 According to another embodiment of the present invention, the real-time cell monitoring system is a system capable of simultaneously measuring local field potential and oxygen consumption rate. Specifically, the local field potential can be measured at the lower part of the cell chip (200), and the oxygen consumption rate can be measured at the upper part of the cell chip (200).

[0104] At this point, the system capable of simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention is substantially the same as the real-time monitoring system according to an embodiment of the present invention, except that the cell electrical signal measurement module (600) includes a local field potential measurement unit (610) instead of an impedance measurement unit (100). Therefore, for the sake of brevity, detailed descriptions of the repetitive configuration will be omitted.

[0105] The cell electrical signal measurement module (600) can measure signals with a certain pulsation, such as myocardial cells, or measure various electrical signal changes of nerve cells. The cell electrical signal measurement module (600) includes a local field potential measurement unit (610) and a controller (620).

[0106] The local field potential measuring unit (610) includes a signal amplifier (611) and an analog-to-digital converter (612).

[0107] The signal amplifier (611) can be electrically connected to the common electrode (221) and the working electrode (222) to measure the local field potential between the common electrode (221) and the working electrode (222). The analog-to-digital converter (612) can be electrically connected to the signal amplifier (611) and the controller (620) to output the measured local field potential as a digital signal.

[0108] Additionally, according to another embodiment of the present invention, a system capable of simultaneously measuring local field potential and oxygen consumption rate may further include a processor (not shown) electrically connected to a cell electrical signal measurement module (600) and an oxygen consumption rate measurement module (500). The processor can analyze the state of cells by utilizing local field potential data measured by the cell electrical signal measurement module (600) and oxygen consumption rate data measured by the oxygen consumption rate measurement module (500).

[0109] At this point, the system capable of simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention can be implemented by the processor and multiple objects, or by a single object. In the case of implementation by multiple objects, a series of processes for analyzing cell state can be executed separately, and these processes can transmit and receive various data via wired or wireless communication networks.

[0110] Figure 11 This is a diagram illustrating a cell electrical signal measurement module constituting a real-time cell monitoring system according to yet another embodiment of the present invention.

[0111] Reference Figure 11 Another embodiment of the present invention is a real-time cell monitoring system that can simultaneously measure cell impedance, local field potential and oxygen consumption rate. Specifically, cell impedance and local field potential can be measured at the lower part of the cell chip (200), and oxygen consumption rate can be measured at the upper part of the cell chip (200).

[0112] At this point, compared with the real-time monitoring system according to one embodiment of the present invention, the system according to another embodiment of the present invention capable of simultaneously measuring cell impedance, local field potential and oxygen consumption rate is substantially the same except that the cell electrical signal measurement module (700) also includes a local field potential measurement unit (720). Therefore, for the sake of brevity, detailed descriptions of the repetitive configuration will be omitted.

[0113] The cell electrical signal measurement module (700) can simultaneously measure cell impedance and local field potential. The cell electrical signal measurement module (700) includes an impedance measurement unit (710), a local field potential measurement unit (720), and a controller (730).

[0114] The impedance measurement unit (710) may include a signal generator (711), a digital-to-analog converter (712), an analog-to-digital converter (713), a first signal amplifier (714a), a second signal amplifier (714b), and a multiplexer (715). Descriptions of each component are consistent with those in... Figure 3 The description is the same as that in the previous text, so its specific details will be omitted.

[0115] The local field potential measuring unit (720) may include a signal amplifier (721), a filter (722), and an analog-to-digital converter (723), and can measure the local field potential between the common electrode (221) and the working electrode (222) to output a first digital signal. On the other hand, with Figure 8 In comparison, the local field potential measuring unit (720) includes a filter (722) in addition to the above. Figure 8 They are essentially the same, so a detailed explanation of the repetition is omitted.

[0116] According to another embodiment of the present invention, a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate can simultaneously operate an impedance measurement unit (710) and a local field potential measurement unit (720). In this case, the effect of the AC power applied to the common electrode (221) for measuring impedance can be reflected in the local field potential measurement result. That is, the first digital signal output by the local field potential measurement unit (720) includes noise caused by the AC power supply. According to another embodiment of the present invention, in order to obtain a second digital signal with the noise removed, an additional process is performed in the controller (730) of the system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate.

[0117] The controller (730) can convert the first digital signal output from the local field potential measuring unit (720) to the frequency domain, filter the noise, and then convert it to the time domain to obtain the second digital signal. The process of converting the first digital signal to the frequency domain can be performed by using the Fourier transform of the following mathematical formula 2, and the process of filtering the noise can be performed by using a low-pass filter (LPF) or a notch filter to filter the signal converted to the frequency domain.

[0118] [Mathematical Expression 3]

[0119] Where X(k) represents the signal in the frequency domain, x(n) represents the signal in the time domain, k represents the frequency, n represents the time, e represents the natural constant, i represents the imaginary unit, and ² represents the partial sum of the sequence.

[0120] Furthermore, according to another embodiment of the present invention, the system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate may further include a processor (not shown) electrically connected to a cell electrical signal measurement module (700) and an oxygen consumption rate measurement module (500). The processor can analyze the cell state by utilizing impedance data and local field potential data measured by the cell electrical signal measurement module (700) and oxygen consumption rate data measured by the oxygen consumption rate measurement module (500).

[0121] At this point, the system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention can be implemented by the processor and multiple objects, or by a single object. In the case of implementation by multiple objects, a series of processes for analyzing cell state can be executed separately, and these processes can transmit and receive various data via wired or wireless communication networks.

[0122] The embodiments of the present invention described above are merely exemplary. Those skilled in the art should understand that the scope of protection of the present invention can include various modifications and equivalents derived therefrom.

[0123] Explanation of reference numerals in the attached figures 1: Real-time cell monitoring system; 100, 600, 700: Cell electrophysiological signal measurement module; 110, 710: Impedance measurement section; 200: Cell chip; 300: Boot module; 400: Board module; 500: Oxygen consumption rate measurement module.

Claims

1. A system for simultaneously measuring cell impedance and oxygen consumption rate in a real-time cell monitoring system, wherein, The system includes: A cell electrical signal measurement module, which includes an impedance measurement unit and a controller; A cell chip, which is fastened to the cell electrical signal measurement module, includes: a hole having a receiving portion open at the top to accommodate cells and culture medium; and a substrate disposed at the lower part of the hole, and having a common electrode and a working electrode formed thereon. A guiding module, which is fastened to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip, and has an opening to expose the upper end of the hole; A plate module, fastened to the upper part of the guide module, and including a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and The oxygen consumption rate measuring module is fixed to the upper part of the plate module and includes a light-emitting part that sends light to the oxygen-responsive substance and a sensor part that receives the optical signal generated from the oxygen-responsive substance.

2. The system according to claim 1, wherein, The impedance measuring unit is electrically connected to the common electrode and the working electrode, and measures the impedance between the common electrode and the working electrode by applying an AC signal to the common electrode.

3. The system according to claim 1, wherein, The probe is located inside the receiving portion of the hole.

4. The system according to claim 3, wherein, The oxygen consumption rate measuring module also includes a driving component that moves the plate module up and down. As the plate module moves up and down, the distance between the probe and the bottom of the receiving part of the hole changes.

5. The system according to claim 1, wherein, The system also includes a processor electrically connected to the cell electrophysiological signal measurement module and the oxygen consumption rate measurement module. The processor analyzes the state of the cells by using impedance data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

6. A system for simultaneously measuring local field potential and oxygen consumption rate in a real-time cell monitoring system, wherein, The system includes: A cell electrical signal measurement module, which includes a local field potential measurement unit and a controller; A cell chip, which is fastened to the cell electrical signal measurement module, includes: a hole having a receiving portion open at the top to accommodate cells and culture medium; and a substrate disposed at the lower part of the hole, and having a common electrode and a working electrode formed thereon. A guiding module, which is fastened to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip, and has an opening to expose the upper end of the hole; A plate module, fastened to the upper part of the guide module, and including a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and The oxygen consumption rate measuring module is fixed to the upper part of the plate module and includes a light-emitting part that sends light to the oxygen-responsive substance and a sensor part that receives the optical signal generated from the oxygen-responsive substance.

7. The system according to claim 6, wherein, The local field potential measuring unit is electrically connected to the common electrode and the working electrode, and measures the local field potential between the common electrode and the working electrode.

8. The system according to claim 6, wherein, The probe is located inside the receiving portion of the hole.

9. The system according to claim 8, wherein, The oxygen consumption rate measuring module also includes a driving component that moves the plate module up and down. As the plate module moves up and down, the distance between the probe and the bottom of the receiving part of the hole changes.

10. The system according to claim 6, wherein, The system also includes a processor electrically connected to the cell electrophysiological signal measurement module and the oxygen consumption rate measurement module. The processor analyzes the state of the cells by using local field potential data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

11. A system for simultaneously measuring cell impedance, local field potential, and oxygen consumption rate in a real-time cell monitoring system, wherein, The system includes: A cell electrical signal measurement module, which includes an impedance measurement unit, a local field potential measurement unit, and a controller; A cell chip, which is fastened to the cell electrical signal measurement module, includes: a hole having a receiving portion open at the top to accommodate cells and culture medium; and a substrate disposed at the lower part of the hole, and having a common electrode and a working electrode formed thereon. A guiding module, which is fastened to the cell electrical signal measurement module to cover the peripheral area of ​​the cell chip, and has an opening to expose the upper end of the hole; A plate module, fastened to the upper part of the guide module, and including a probe extending along the depth direction of the receiving portion and having an oxygen-responsive substance attached to its lower part; and The oxygen consumption rate measuring module is fixed to the upper part of the plate module and includes a light-emitting part that sends light to the oxygen-responsive substance and a sensor part that receives the optical signal generated from the oxygen-responsive substance.

12. The system according to claim 11, wherein, The impedance measuring unit and the local field potential measuring unit are electrically connected to the common electrode and the working electrode. The impedance measurement unit measures the impedance between the common electrode and the working electrode by applying an AC signal to the common electrode, and the local field potential measurement unit measures the local field potential between the common electrode and the working electrode.

13. The system according to claim 11, wherein, The probe is located inside the receiving portion of the hole.

14. The system according to claim 13, wherein, The oxygen consumption rate measuring module also includes a driving component that moves the plate module up and down. As the plate module moves up and down, the distance between the probe and the bottom of the receiving part of the hole changes.

15. The system according to claim 11, wherein, The system also includes a processor electrically connected to the cell electrophysiological signal measurement module and the oxygen consumption rate measurement module. The processor analyzes the state of the cells by using impedance data and local field potential data measured by the cell electrical signal measurement module and oxygen consumption rate data measured by the oxygen consumption rate measurement module.

Images