Real-time cell monitoring system

Abstract

The present invention provides a real-time cell monitoring system capable of improving the accuracy and convenience of real-time cell analysis by simultaneously measuring various electrical signals and an oxygen consumption rate of a cell in a non-destructive manner.

Description

Real-time cell monitoring system

[0001] The present invention relates to a real-time cell monitoring system, and more specifically, to a real-time cell monitoring system capable of improving the accuracy and convenience of real-time cell analysis by simultaneously measuring various electrical signals and oxygen consumption rates of cells in a non-destructive manner.

[0002] Cell analysis has established itself as an essential element in modern biological and medical research. In particular, accurately analyzing the state of cells—a critical biological process in which cells transform into mature forms with specific functions—plays a vital role in various studies and experiments, including disease diagnosis, the development of treatments, and the evaluation of drug efficacy.

[0003] Conventional cell analysis utilized methods such as microscopic observation and biochemical analysis, while cell differentiation evaluation employed methods including gene expression analysis, protein expression profiling, immunofluorescence, and flow cytometry. However, these methods not only made it difficult to monitor dynamic cellular changes in real time but also had limitations in terms of cell contamination, accuracy, cost, and time.

[0004] Recently, various non-destructive real-time cell analysis technologies, such as Electrical Cell-Substrate Impedance Sensing (ECIS), Local Field Potential (LPP) measurement, and Oxygen Consumption Rate (OCR) measurement, are being researched. However, while these technologies can analyze the functional state and changes of cells in real time, they are limited in that most are restricted to measuring specific data individually.

[0005] Therefore, there is a need to develop a new system capable of more accurately and in real-time monitoring and analysis of cell conditions, such as cell metabolism, cell culture, cell differentiation, and drug screening.

[0006] The technical problem that the present invention aims to solve is to provide a real-time cell monitoring system capable of improving the accuracy and convenience of real-time cell analysis by simultaneously measuring various electrical signals and oxygen consumption rates of cells using a non-destructive method.

[0007] The technical problems of the present invention are not limited to those mentioned above, and other unmentioned technical problems will be clearly understood by a person skilled in the art from the description below.

[0008] A real-time cell monitoring system for simultaneously measuring cell impedance and oxygen consumption rate according to an embodiment of the present invention for achieving the above technical problem comprises: a cell electrical signal measuring module including an impedance measuring unit and a controller; a cell chip including a well connected to the cell electrical signal measuring module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed below the well having a common electrode and an operating electrode formed thereon; a guide module connected to the cell electrical signal measuring module to cover a peripheral area of ​​the cell chip and having an opening formed so that the top of the well is exposed; a plate module connected to the upper part of the guide module and including a probe extending in the depth direction of the receiving portion and having an oxygen-sensitive material attached to its lower part; and an oxygen consumption rate measuring module connected to the upper part of the plate module and including a light-emitting portion that transmits light to the oxygen-sensitive material and a sensor portion that receives an optical signal generated from the oxygen-sensitive material.

[0009] The impedance measuring unit is 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 alternating current signal to the common electrode.

[0010] The above probe may be located inside the receiving portion of the well.

[0011] The oxygen consumption rate measuring module may further include a driving member that moves the plate module up and down, and a change in the distance between the probe and the bottom of the well's receiving portion may occur depending on the up and down movement of the plate module.

[0012] The above system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and the processor can analyze the state of the cell 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] A real-time cell monitoring system for simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention for achieving the above technical problem comprises: a cell electrical signal measuring module including a local field potential measuring unit and a controller; a cell chip including a well connected to the cell electrical signal measuring module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed below the well having a common electrode and an operating electrode formed thereon; a guide module connected to the cell electrical signal measuring module to cover a peripheral area of ​​the cell chip and having an opening formed so that the top of the well is exposed; a plate module connected to the upper part of the guide module and including a probe extending in the depth direction of the receiving portion and having an oxygen-sensitive material attached to its lower part; and an oxygen consumption rate measuring module connected to the upper part of the plate module and including a light-emitting portion that transmits light to the oxygen-sensitive material and a sensor portion that receives an optical signal generated from the oxygen-sensitive material.

[0014] The local field potential measuring unit is 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 above probe may be located inside the receiving portion of the well.

[0016] The oxygen consumption rate measuring module may further include a driving member that moves the plate module up and down, and a change in the distance between the probe and the bottom of the well's receiving portion may occur depending on the up and down movement of the plate module.

[0017] The above system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and the processor can analyze the state of the cell 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.

[0018] A real-time cell monitoring system for simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention for achieving the above technical problem comprises: a cell electrical signal measuring module including an impedance measuring unit, a local field potential measuring unit, and a controller; a cell chip including a well connected to the cell electrical signal measuring module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed below the well having a common electrode and an operating electrode formed thereon; a guide module connected to the cell electrical signal measuring module to cover a peripheral area of ​​the cell chip and having an opening formed so that the top of the well is exposed; a plate module connected to the upper part of the guide module and including a probe extending in the depth direction of the receiving portion and having an oxygen-sensitive material attached to its lower part; and an oxygen consumption rate measuring module connected to the upper part of the plate module and including a light-emitting unit that transmits light to the oxygen-sensitive material and a sensor unit that receives an optical signal generated from the oxygen-sensitive material.

[0019] The impedance measuring unit and the local field potential measuring unit are electrically connected to the common electrode and the working electrode, and the impedance measuring unit measures the impedance between the common electrode and the working electrode by applying an alternating current signal to the common electrode, and the local field potential measuring unit can measure the local field potential between the common electrode and the working electrode.

[0020] The above probe may be located inside the receiving portion of the well.

[0021] The oxygen consumption rate measuring module may further include a driving member that moves the plate module up and down, and a change in the distance between the probe and the bottom of the well's receiving portion may occur depending on the up and down movement of the plate module.

[0022] The above system may further include a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and the processor may analyze the state of the cell 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.

[0023] According to the present invention as described above, there is an effect of simultaneously monitoring 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, by utilizing simultaneously measured data on cell impedance, local field potential, or oxygen consumption rate, it is possible to more accurately analyze the state of cells, such as cell metabolism, cell culture, cell differentiation, and drug screening.

[0025] The effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by a person skilled in the art from the description below.

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

[0027] Figure 2 is an exploded perspective view schematically illustrating the configuration of the real-time cell monitoring system of Figure 1.

[0028] FIGS. 3(a) to 3(c) are drawings for explaining a cell electrical signal measurement module constituting a real-time cell monitoring system according to one embodiment of the present invention.

[0029] FIG. 4 is a diagram showing an example of a cell chip constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0030] Figure 5 is a diagram showing an example of a substrate constituting the cell chip of Figure 4.

[0031] FIGS. 6(a) and FIGS. 6(b) are drawings showing an example of a guide module constituting a real-time cell monitoring system according to an embodiment of the present invention.

[0032] FIG. 7 is a drawing showing an 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] FIG. 8 is a graph showing impedance results measured using a real-time cell monitoring system according to one embodiment of the present invention.

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

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

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

[0037] The advantages and features of the present invention and the methods for achieving them will become clear by referring to the embodiments described below in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below but may be implemented in various different forms. These embodiments are provided merely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined only by the scope of the claims.

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

[0039] The terms used in this invention have been selected based on currently widely used general terms, taking into account their functions within the invention; however, these terms may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc. Additionally, in specific cases, terms have been arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0040] Throughout the specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components. Furthermore, terms such as "part," "module," and "unit" used in the specification refer to a unit that processes at least one function or operation and may be implemented as software, hardware components such as FPGAs or ASICs, or a combination of software and hardware. However, the terms "part," "module," and "unit" are not limited to software or hardware. "Part," "module," and "unit" may be configured to reside in an addressable storage medium or configured to operate one or more processors. Therefore, as an example, terms such as “part,” “module,” and “unit” include components such as software components, object-oriented software components, class components, and task components, as well as processes, functions, attributes, procedures, subroutines, segments of program code, drivers, firmware, microcode, circuits, data, databases, data structures, tables, arrays, and variables.

[0041] Below, embodiments of the present invention are described in detail with reference to the attached drawings so that those skilled in the art can easily implement the invention. Additionally, parts of the drawings that are irrelevant to the description are omitted to clearly explain the invention.

[0042] Terms including ordinal numbers, such as “first,” “second,” etc., may be used to describe various components, but the components are not limited by the terms. The terms are used solely for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the first component may be named the second component, and similarly, the second component may be named the first component. The term “and / or” includes a combination of multiple related items or any one of the multiple related items.

[0043]

[0044] FIG. 1 is a schematic perspective view of a real-time cell monitoring system according to an embodiment of the present invention, FIG. 2 is an exploded perspective view schematically illustrating the configuration of the real-time cell monitoring system of FIG. 1, FIG. 3(a) to FIG. 3(c) are drawings for explaining a cell electrical signal measurement module constituting a real-time cell monitoring system according to an embodiment of the present invention, FIG. 4 is a drawing showing an embodiment of a cell chip constituting a real-time cell monitoring system according to an embodiment of the present invention, FIG. 5 is a drawing showing an embodiment of a substrate constituting the cell chip of FIG. 4, FIG. 6(a) and FIG. 6(b) are drawings showing an embodiment of a guide module constituting a real-time cell monitoring system according to an embodiment of the present invention, and FIG. 7 is a drawing showing an 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.

[0045]

[0046] Referring to FIGS. 1 and 2, a real-time cell monitoring system (1, hereinafter also referred to as the ‘system’) according to one embodiment of the present invention 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 one embodiment of the present invention is a system (1) capable of simultaneously measuring cell impedance (hereinafter also referred to as ‘impedance’ or ‘impedance data’) and oxygen consumption rate, specifically, measuring cell impedance at the bottom of the cell chip (200) and measuring oxygen consumption rate at the top of the cell chip (200).

[0047] Additionally, the system (1) may further include a processor (not shown) electrically connected to a cell electrical signal measurement module (100) and an oxygen consumption rate measurement module (500). The processor can analyze the state of the cell 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).

[0048] At this time, the system (1) may be implemented with the processor and a plurality of objects or as a single object. When implemented with a plurality of objects, a series of processes for analyzing the state of the cell can be divided and performed, and they can exchange various data through a wired communication network or a wireless communication network.

[0049] Referring to FIG. 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 role of measuring the impedance generated during the process of culturing cells or processing cultured cells by applying an alternating current signal to the cell chip (200) and generating impedance data. Specifically, when an alternating current signal is applied during the process of culturing cells in the well (210) of the cell chip (200), as the number of cells increases due to cell culture, the area covering the common electrode (221) and the working electrode (222) placed at the bottom of the well (210) increases, and the current is physically obstructed, causing the impedance to increase. Conversely, when an alternating current signal is applied during the process of treating cells cultured in the well (210) of the cell chip (200) with drugs, the impedance decreases as the cell becomes smaller due to the cell treatment, and the area covering the common electrode (221) and the working electrode (222) placed at the bottom of the well (210) decreases.

[0050] Referring to FIG. 3(a), in one embodiment of the present invention, the impedance measuring unit (110) includes a signal generator (111), a digital-to-analog converter (112), and an analog-to-digital converter (113).

[0051] The signal generator (111) can generate an alternating current signal controlled by the controller (120) described later, and can be configured together with the controller (120) or provided as a separate power device and connected to the controller (120).

[0052] A digital-to-analog converter (112, digital-to-analog converter; DAC) is electrically connected to a common electrode (221) corresponding to each cell chip (200), and can convert an alternating current signal generated from a signal generator (111) into an analog input signal and then apply it to the common electrode (221) corresponding to each cell chip (200).

[0053] An analog-to-digital converter (113, analog-to-digital converter; ADC) is electrically connected to an operating electrode (222) corresponding to each cell chip (200) and can receive an analog response signal generated between a common electrode (221) and an operating electrode (222), convert it into a digital signal, and then transmit it to a controller (120). Of course, the digital signal converted by the analog-to-digital converter (113) can be transmitted directly to the controller (120) or transmitted to the controller (120) after passing through additional configurations at a later stage.

[0054] Referring to FIG. 3(b), in another embodiment of the present invention, the impedance measuring unit (110) may further include a first signal amplifier (114a, amplifier; AMP) that amplifies an analog input signal at the output of a digital-to-analog converter (112) and a second signal amplifier (114b) that amplifies an analog response signal at the output of an analog-to-digital converter (113). In this case, the first signal amplifier (114a) or the second signal amplifier (144b) may be a plurality of.

[0055] Referring to FIG. 3(c), in another embodiment of the present invention, the impedance measuring unit (110) may further include a multiplexer (115) that includes a plurality of input channels and one output channel at the front end of the second signal amplifier (114b) and connects one of the plurality of input channels to the output channel. In one embodiment of the present invention, when the cell chip (200) is composed of a plurality of wells (210), a common electrode (221) and an operating electrode (222) are disposed at the bottom of the receiving portion (212) of each well (210), and the multiplexer (115) may receive all different analog response signals generated between the common electrode (221) and the operating electrode (222) corresponding to each well (210) and selectively output them.

[0056] The controller (120) can control the overall operation of the impedance measuring unit (110), and specifically, can control the frequency of the alternating current signal applied to the cell chip (200) through the impedance measuring unit (110) and the impedance measurement time.

[0057] Additionally, the controller (120) can receive information about the impedance measured by the impedance measuring unit (110) and generate impedance data, and can transmit the generated impedance data to a processor (not shown) described later.

[0058] Specifically, the controller (120) can generate impedance data among resistive data, capacitive data, magnitude data, phase data, and combinations thereof. More specifically, the controller (120) receives information about impedance provided in the form of a sum of a resistance value, which is a real part, and a reactance value, which is an imaginary part, from the impedance measuring unit (110), generates resistive data using the resistance value, generates capacitive data using the reactance value, which is an imaginary part, and generates magnitude data using both the resistance value, which is a real part, and the reactance value, which is an imaginary part. Meanwhile, it goes without saying that the controller (120) can generate impedance data for the cell and impedance data for the culture medium, respectively.

[0059] In addition, the controller (120) can generate different data depending on the control operation for the impedance measuring unit (110).

[0060] For example, the controller (120) can control the frequency of the AC signal to a single value during the time the impedance is measured in the impedance measuring unit (110), and the controller (120) can generate impedance data over time.

[0061] As another example, the controller (120) can control the frequency of the AC signal to a plurality of different values ​​between the minimum frequency and the maximum frequency during the time the impedance is measured in the impedance measuring unit (110), and the controller (120) can generate impedance data by time and / or frequency.

[0062] As another example, the controller (120) can control the frequency of the AC signal to a plurality of different values ​​between the minimum frequency and the maximum frequency during the time the impedance is measured in the impedance measuring unit (110), and at the same time, control the measurement time differently for each frequency of the AC signal, and at this time, the controller (120) can generate impedance data by time and / or by frequency. Through this, the accuracy of the impedance data can be improved as the required measurement time increases as the number of frequencies of the AC signal applied to the cell chip (200) increases and the frequency value decreases.

[0063] In this way, the system (1) of the present invention can control the operation of the impedance measuring unit (110) in various ways according to the purpose of measuring cell impedance, and the controller (120) can generate different data according to the control operation, thereby greatly improving the convenience of cell analysis.

[0064] Referring to FIGS. 4 and 5, the cell chip (200) comprises a well (210) for culturing cells or processing cultured cells, and a substrate (220) having a common electrode (221) and an operating electrode (222) formed thereon. In one example, the well (210) may be placed on the substrate (220), whereby the well (210) may be fixed by a heat compression method at 60 to 70°C and 30 to 30 psi on a substrate (120) to which double-sided tape made of polyethylene, silicone, polyurethane, or polyester material is attached. However, this is not limited thereto, and the well (210) may be fixed on the substrate (220) by various methods.

[0065] The well (210) is configured for culturing cells or processing cultured cells, and is preferably made of a transparent biocompatible material. The biocompatible material may be a transparent plastic such as PDMS, PMMA, PET, or PC, but is not limited thereto and may be made of transparent glass, etc.

[0066] The well (210) has a receiving section (212) that can accommodate cells and culture medium, with the top open by a partition (211), and the cross-section of the receiving section (212) can be formed in various shapes such as a circle or a square. Additionally, the well (210) can be formed as a single receiving section (212) in which a space is formed inside by a partition (211) arranged in a vertical direction, and the receiving sections (212) can be arranged adjacently in rows (horizontal direction) and columns (vertical direction) or spaced apart by the partition (211). At this time, the number of receiving sections (212) can be determined in various ways, such as 4, 6, 7, 12, 16, 24, 48, 96, 128, etc.

[0067] The substrate (220) supports the well (210) and simultaneously receives an alternating current signal from the cell electrical signal measurement module (100). The substrate (220) is preferably a non-conductive substrate made of a transparent material such as glass or plastic to optically monitor the cell culture or cell processing state.

[0068] A plurality of electrodes are formed on the surface of the substrate (220), which are divided into a common electrode (221) and an operating electrode (222) spaced apart from the common electrode (221). A transmission line (223) and a terminal pad (224) may be further formed together with the common electrode (221) and the operating electrode (222). Meanwhile, in another embodiment of the present invention, a stimulation electrode (not shown) may be further formed on the surface of the substrate (220) at a position spaced apart from the common electrode (221) and the operating electrode (222).

[0069] The common electrode (221) may include a body portion and a portion of the horizontal side of the body portion that are recessed inwardly. In this case, the recessed portion is preferably positioned at the bottom of the receiving portion (212) of the well (210) and may be arranged in multiple numbers spaced apart at regular intervals from the horizontal side of the body portion. Additionally, the recessed portion is preferably in the shape of a semicircular arc with a certain curvature, but is not limited thereto and may be in the shape of a square or triangle recessed inwardly.

[0070] The operating electrode (222) may be disposed within the recess formed in the common electrode (221) and may be formed in various shapes depending on the shape of the recess. For example, if the recess is a semicircular arc shape having a certain curvature, the operating electrode (222) may be a disk shape corresponding thereto.

[0071] The working electrode (222) is spaced apart from the common electrode (221) and can form an impedance according to an alternating current signal applied to the common electrode (221) from the impedance measurement module (100). More specifically, as it is spaced apart from the common electrode (221) at the bottom of the receiving portion (212) of the well (210), real-time impedance changes resulting from cell culture or cell treatment in the receiving portion (212) of the well (210) can be collected in a non-destructive manner and analyzed.

[0072] The transmission line (223) serves to connect the common electrode (221) and the operating electrode (222) with the terminal pad (224), and the terminal pad (224) is electrically connected to the impedance measuring unit (110) to receive an alternating current signal and can apply the alternating current signal to the common electrode (221) through the transmission line (223).

[0073] In the case where a plurality of recesses and a plurality of operating electrodes (222) are formed in a common electrode (221) as in one embodiment of the present invention (see FIG. 5), the transmission lines (223) connected to each operating electrode (222) can be formed with different widths considering the coupling and parasitic capacitance generated by adjacent transmission lines (223) and connected to the terminal pad (224), and together with the distance from the terminal pad (224), they can be formed with different spacing from adjacent transmission lines (223). More specifically, it is preferable that the transmission lines (223) connected to operating electrodes (222) far from the terminal pad (224) are formed with a wide width, and the transmission lines (223) are formed with a narrower width as they are connected to operating electrodes (222) closer to the terminal pad (224). Additionally, it is preferable that the transmission line (223) connected to the operating electrode (222) far from the terminal pad (224) has a wider gap with the adjacent transmission line (223) compared to the transmission line (223) connected to the operating electrode (222) close to the terminal pad (224), thereby minimizing well variation and increasing the reliability of cell analysis. At this time, the width and length of the transmission line (223) can be determined according to the theoretical sheet resistance calculation value according to Equation 1.

[0074]

[0075] Here, R sheet ε represents the sheet resistance of the transmission line (223), ε represents the resistivity of the material of the transmission line (223), l represents the length of the current flow between the working electrode (222) and the terminal pad (224), w represents the width of the transmission line (223), and h represents the height of the transmission line (223).

[0076] The common electrode (221), operating electrode (222), transmission line (223), and terminal pad (224) can be formed as transparent, reflective, or semi-transparent types, and can also be formed of a biocompatible conductive material.

[0077] For example, the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) may be formed by depositing a transparent metal oxide, such as ITO (indium tin oxide), IZO (indium zinc oxide), ZnO (zinc oxide), or ITZO (indium thin zinc oxide), onto the substrate (220) and then patterning it, or by depositing a transparent metal oxide and a photoresist together and then etching it, or by a laser patterning method. In this case, the well (210) and the substrate (220), as well as the common electrode (221), working electrode (222), transmission line (223), and terminal pad (224) formed on the substrate (220), are all formed from a transparent material, which has an advantageous effect for optically monitoring the cell culture or cell processing status.

[0078] Meanwhile, when the common electrode (221) and the working electrode (222) are formed of a transparent material, any one of the following nanomaterials may be deposited on the surface: gold nanoparticles (Au nanoparticles, AuNPs), quantum dot (QD) reduced graphene oxide (rGO), poly(3,4-ethylenedioxythiophene)-polystyrene sulfonate (PEDOT:PSS), polyaniline (PANI), carbon nanotubes (CNT), and multi-walled carbon nanotubes (MWCNT). In this case, it is preferable that the nanomaterial be a biocompatible conductive material, but the nanomaterial is not limited to the nanomaterials exemplified above as long as it is a biocompatible conductive material. As the interfacial impedance of the common electrode or working electrode is lowered through the above nanomaterial deposition, the sensitivity and accuracy of electrical impedance measurement can be further improved.

[0079] 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, which are 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) all have improved productivity and cost-effectiveness compared to forming them from a transparent material.

[0080] Referring to FIGS. 2 and FIGS. 6, the guide module (300) is connected to the cell electrical signal measurement module (100) and performs the role of guiding the placement position of the plate module (400). Specifically, it is preferable that the guide module (300) be connected to the cell electrical signal measurement module (100) so as to cover the surrounding area of ​​the cell chip (200).

[0081] The guide module (300) has an opening (310) formed so that the upper part of the well (210) of the cell chip (200) is exposed when connected to the cell electrical signal measurement module (100). Additionally, to ensure stability when connected to the cell electrical signal measurement module (100), a connecting projection (320) may be further formed on the side of the lower surface of the guide module (300). At this time, the cell electrical signal measurement module (100) may have a connecting groove (130) formed so that the connecting projection (320) of the guide module (300) is inserted, and a fixing member (140) for fixing the guide module (300) may be further formed.

[0082] Additionally, the guide module (300) may be divided into an upper frame in which an opening (310) is formed and a lower frame in which a fastening projection (320) is formed. At this time, the lower frame may be formed wider than the upper frame so that a space is formed between the end of the upper frame and the end of the lower frame, and the plate module (400) described later may be fastened into the space.

[0083] Referring to FIGS. 2 and FIGS. 7, the plate module (400) is attached to the upper part of the guide module (300), and the oxygen consumption rate measuring module (500) is attached to the upper part of the plate module (400). In one embodiment of the present invention, the plate module (400) and the oxygen consumption rate measuring module (500) can measure the oxygen concentration of the culture medium contained in the well (210) of the cell chip (200) over time on the upper part of the cell chip (200), and then measure the oxygen consumption rate (OCR) using the measured oxygen concentration and the following Equation 1.

[0084]

[0085] Here, d[O2] / dt represents the rate of change of oxygen concentration per unit time, and V represents the volume of the culture medium.

[0086] The plate module (400) includes a probe (410) and a frame (420).

[0087] The probe (410) may be in the form of a rod having a certain length and extending in the depth direction of the receiving portion (212) of the well (210). At this time, the probe (410) may be formed to be located inside the receiving portion (212) of the well (210). Specifically, the probe (410) is preferably formed in the lower region of the frame (420) corresponding to the opening (310) of the guide module (300) and is formed to be located in the center inside the receiving portion (212) of the well (210), and it is even more preferable that it be formed spaced apart from the bottom of the receiving portion of the well (210) at a certain distance.

[0088] Additionally, the probe (410) may have an oxygen-sensitive material attached to its lower portion for measuring oxygen concentration. In this case, the oxygen-sensitive material may be any one of metal complexes, organic fluorophores, optoelectronic materials, nanomaterials, optical fiber materials, silica gel, and polymer matrix. However, it is not limited to these, and any material capable of generating or changing a signal in response to oxygen concentration may be used as an oxygen-sensitive material.

[0089] Additionally, the probes (410) may be formed in multiple numbers equal to the number of receiving portions (212), and as in one embodiment of the present invention, when 16 receiving portions are formed on one side and 16 on the other side, for a total of 32, the probes (410) may also be formed in a total of 32 numbers.

[0090] The frame (420) may be divided into a main frame (421) and a support frame (422), and the main frame (421) and the support frame (422) may be integral or separate. A portion of the lower inner surface 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), thereby allowing the plate module (400) to be stably supported on the upper part of the guide module (300).

[0091] Additionally, the frame (420) may be able to move up and down in the vertical direction by external pressure, and for this purpose, the support frame (422) of one embodiment of the present invention may be made of a flexible or elastic member. However, it is not limited thereto, and the frame (420) may be able to move up and down in the vertical direction by external pressure by forming a separate flexible member on the main frame (421).

[0092] At this time, the probe (410) moves up and down in the vertical direction inside the receiving portion (212) of the well (210) according to the vertical movement of the frame (420), and a change may occur in the distance between the probe (410) and the bottom of the receiving portion (212) of the well (210). At this time, it is preferable that the probe (410) does not come into contact with the bottom of the receiving portion (212) of the well (210) even when moving downward inside the receiving portion (212) of the well (210), but is spaced apart at a certain distance.

[0093] Additionally, the plate module (400) may further include an optical lens (430). The optical lens (430) is positioned inside the main frame (421), and more specifically, may be positioned in an area corresponding to the location where the probe (410) and the light-emitting part (510) and sensor part (520) of the oxygen consumption rate measurement module (500) described later are formed.

[0094] The optical lens (430) can focus the light transmitted from the light-emitting part (510) of the oxygen consumption rate measurement module (500) onto the oxygen-sensitive material of the probe (410) to more accurately measure the optical signal that changes according to the oxygen concentration, and can also control the path of the optical signal generated from the oxygen-sensitive material to effectively focus it to the sensor part (520) to improve sensitivity.

[0095] Additionally, the plate module (400) may further include a temperature sensor (not shown), an atmospheric pressure sensor (not shown), and / or a humidity sensor (not shown). The temperature sensor, atmospheric pressure sensor, and humidity sensor may be positioned adjacent to the probe (410) to measure the temperature, atmospheric pressure, and / or humidity inside the receiving portion (212) of the well (210), and the accuracy of the oxygen consumption rate (OCR) may be further improved using the measured temperature value, atmospheric pressure value, and / or humidity value.

[0096] 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 positioned on the upper part of the oxygen consumption rate measuring module (500), but are not limited thereto and may be positioned on the lower part in a manner that contacts the upper part of the plate module (400).

[0097] The light-emitting part (510) transmits light using an oxygen-sensitive material attached to the lower part of the probe (410), and may be a light-emitting diode (LED), but is not limited thereto and may use various devices capable of emitting light.

[0098] The sensor unit (520) receives an optical signal generated from an oxygen-sensitive material attached to the lower part of the probe (410), and may be an optical sensor, but is not limited thereto, and may use various elements capable of detecting light.

[0099] Additionally, the oxygen consumption rate measuring module (500) may further include a driving member (not shown) that moves the plate module (400) up and down, and the plate module (400) moves up and down in the up and down direction by the operation of the driving member, and accordingly, a change may occur in the distance between the probe (410) and the bottom of the receiving portion (212) of the well (210).

[0100] At this time, the driving member may include an actuator and a driving pin connected to the actuator, and the actuator may be positioned on one side or / and the other side of the oxygen consumption rate measuring module (500), and the driving pin may be positioned to be connected to the support frame (422) of the plate module (400). However, the driving member is not limited thereto and may be formed in various configurations capable of moving the plate module (400) up and down and may be positioned in various ways.

[0101] Additionally, the oxygen consumption rate measurement module (500) may further include a control unit (not shown) and a communication unit (not shown). The control unit can control the amount of light from the light-emitting unit (510) and can measure (calculate) the oxygen concentration and oxygen consumption rate using an 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 described above via wired or wireless means.

[0102] FIG. 8 is a graph showing impedance results measured using a real-time cell monitoring system according to one embodiment of the present invention, and FIG. 9 is a graph showing oxygen consumption rate results measured using a real-time cell monitoring system according to one embodiment of the present invention.

[0103] Referring to FIGS. 8 and 9, a system (1) according to one embodiment of the present invention can simultaneously measure cell impedance in a cell electrical signal measurement module (100) and measure oxygen consumption rate in an oxygen consumption rate measurement module (500) under the same cell measurement conditions to generate hourly impedance data and hourly oxygen consumption rate result graphs, respectively. In addition, a system (1) according to one embodiment of the present invention can generate the measured hourly impedance data and hourly oxygen consumption rate result graphs by superimposing them into a single graph. Through this, the present invention has the effect of more accurately analyzing the state of cells, such as cell metabolism, cell culture, cell differentiation, and drug screening.

[0104] FIG. 10 is a diagram illustrating a cell electrical signal measurement module constituting a real-time cell monitoring system according to another embodiment of the present invention.

[0105] Referring to FIG. 10, a real-time cell monitoring system according to another embodiment of the present invention is a system capable of simultaneously measuring local field potential and oxygen consumption rate, specifically measuring local field potential at the bottom of the cell chip (200) and measuring oxygen consumption rate at the top of the cell chip (200).

[0106] At this time, a system capable of simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention is substantially identical to a real-time monitoring system according to one 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 of the specification, a detailed description of the redundant components is omitted.

[0107] The cell electrical signal measurement module (600) can measure signals with a constant beat, such as those of myocardial cells, or measure various electrical signal changes of nerve cells, and the cell electrical signal measurement module (600) includes a local field potential measurement unit (610) and a controller (620).

[0108] The local field potential measuring unit (610) includes a signal amplifier (611) and an analog-to-digital converter (612).

[0109] A signal amplifier (611) is electrically connected to a common electrode (221) and an operating electrode (222) to measure a local field potential between the common electrode (221) and the operating electrode (222), and an analog-to-digital converter (612) is electrically connected to the signal amplifier (611) and a controller (620) to output the measured local field potential as a digital signal.

[0110] Additionally, a system capable of simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention 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 the cell using 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).

[0111] At this time, a system capable of simultaneously measuring local field potential and oxygen consumption rate according to another embodiment of the present invention may be implemented with the processor and a plurality of objects or as a single object. When implemented with a plurality of objects, a series of processes for analyzing the state of a cell may be divided and performed, and these may exchange various data through a wired communication network or a wireless communication network.

[0112] FIG. 11 is a diagram illustrating a cell electrical signal measurement module constituting a real-time cell monitoring system according to another embodiment of the present invention.

[0113] Referring to FIG. 11, a real-time cell monitoring system according to another embodiment of the present invention is a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate, specifically measuring cell impedance and local field potential at the bottom of the cell chip (200) and measuring oxygen consumption rate at the top of the cell chip (200).

[0114] At this time, a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention is substantially identical to a real-time monitoring system according to one embodiment of the present invention, except that the cell electrical signal measurement module (700) further includes a local field potential measurement unit (720). Therefore, for the sake of brevity in the specification, a detailed description of the redundant components is omitted.

[0115] The cell electrical signal measurement module (700) can simultaneously measure cell impedance and local field potential, and the cell electrical signal measurement module (700) includes an impedance measurement unit (710), a local field potential measurement unit (720), and a controller (730).

[0116] The impedance measuring 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). Since the description of each component is as described in FIG. 3, a detailed description will be omitted.

[0117] The local field potential measuring unit (720) may include a signal amplifier (721), a filter (722), and an analog-to-digital converter (723), and may output a first digital signal by measuring the local field potential between the common electrode (221) and the working electrode (222). Meanwhile, since the local field potential measuring unit (720) is substantially the same as that of FIG. 8 except that it further includes a filter (722), a detailed description of the redundant configuration will be omitted.

[0118] A system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention may have an impedance measuring unit (710) and a local field potential measuring unit (720) operating simultaneously. In this case, the influence of the AC power applied to the common electrode (221) for impedance measurement may be reflected in the local field potential measurement result. That is, the first digital signal output by the local field potential measuring unit (720) contains noise caused by the AC power, and a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention performs an additional process in the controller (730) to obtain a second digital signal from which the noise has been removed.

[0119] The controller (730) can obtain a second digital signal by converting the first digital signal output from the local field potential measuring unit (720) into the frequency domain, filtering out noise, and then converting it into the time domain. The process of converting the first digital signal into the frequency domain can be performed by a Fourier transform using the following mathematical formula 2, and the process of filtering out noise can be performed by filtering the signal converted into the frequency domain with a low-frequency communication filter (LPF) or a notch filter.

[0120]

[0121] Here, X(k) is a signal in the frequency domain, x(n) is a signal in the time domain, k is the frequency, n is the time, e is the natural constant, i is the imaginary unit, and ² represents the partial sum of the sequence.

[0122] Additionally, a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention 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 state of the cell using 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).

[0123] At this time, a system capable of simultaneously measuring cell impedance, local field potential, and oxygen consumption rate according to another embodiment of the present invention may be implemented with the processor and a plurality of objects or as a single object. When implemented with a plurality of objects, a series of processes for analyzing the state of the cell may be divided and performed, and these may exchange various data through a wired or wireless communication network.

[0124] The embodiments of the present invention described above are merely exemplary, and the scope of protection of the present invention may include various modifications and equivalents therefrom, as understood by those skilled in the art.

[0125] 1: Real-time cell monitoring system

[0126] 100, 600, 700: Cell electrical signal measurement module

[0127] 110, 710: Impedance measurement section

[0128] 200: Cell Chip

[0129] 300: Guide Module

[0130] 400: Plate Module

[0131] 500: Oxygen consumption rate measurement module

Claims

1. In a real-time cell monitoring system that simultaneously measures cell impedance and oxygen consumption rate, A cell electrical signal measurement module including an impedance measuring unit and a controller; A cell chip comprising a well connected to the cell electrical signal measurement module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed at the bottom of the well and having a common electrode and an operating electrode formed thereon; A guide module connected to the cell electrical signal measurement module to cover the surrounding area of ​​the cell chip, with an opening formed therein to expose the top of the well; A plate module including a probe that is fastened to the upper part of the guide module, extends in the depth direction of the receiving portion, and has an oxygen-sensitive material attached to its lower portion; and A system comprising: an oxygen consumption rate measuring module that is fastened to the upper part of the plate module and includes a light-emitting part that transmits light to the oxygen-sensitive material and a sensor part that receives an optical signal generated from the oxygen-sensitive material.

2. In Claim 1, The above impedance measuring unit is electrically connected to the common electrode and the working electrode, and a system that measures the impedance between the common electrode and the working electrode by applying an alternating current signal to the common electrode.

3. In Claim 1, The above probe is located inside the receiving portion of the well, in a system.

4. In Claim 3, The oxygen consumption rate measuring module further includes a driving member that moves the plate module up and down, and A system in which a change in the distance between the probe and the bottom of the well's receiving portion occurs according to the lifting and lowering movement of the plate module.

5. In Claim 1, The above system further includes a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and The above processor is a system that analyzes the state of a cell 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. In a real-time cell monitoring system that simultaneously measures local field potential and oxygen consumption rate, A cell electrical signal measurement module including a local field potential measuring unit and a controller; A cell chip comprising a well connected to the cell electrical signal measurement module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed at the bottom of the well and having a common electrode and an operating electrode formed thereon; A guide module connected to the cell electrical signal measurement module to cover the surrounding area of ​​the cell chip, with an opening formed therein to expose the top of the well; A plate module including a probe that is fastened to the upper part of the guide module, extends in the depth direction of the receiving portion, and has an oxygen-sensitive material attached to its lower portion; and A system comprising: an oxygen consumption rate measuring module that is fastened to the upper part of the plate module and includes a light-emitting part that transmits light to the oxygen-sensitive material and a sensor part that receives an optical signal generated from the oxygen-sensitive material.

7. In Claim 6, 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, a system.

8. In Claim 6, The above probe is located inside the receiving portion of the well, in a system.

9. In Claim 8, The oxygen consumption rate measuring module further includes a driving member that moves the plate module up and down, and A system in which a change in the distance between the probe and the bottom of the well's receiving portion occurs according to the lifting and lowering movement of the plate module.

10. In Claim 6, The above system further includes a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and The above processor is a system that analyzes the state of a cell 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. In a real-time cell monitoring system for simultaneously measuring cell impedance, local field potential, and oxygen consumption rate, A cell electrical signal measurement module comprising an impedance measuring unit, a local field potential measuring unit, and a controller; A cell chip comprising a well connected to the cell electrical signal measurement module and having a receiving portion with an open top for accommodating cells and culture medium, and a substrate disposed at the bottom of the well and having a common electrode and an operating electrode formed thereon; A guide module connected to the cell electrical signal measurement module to cover the surrounding area of ​​the cell chip, with an opening formed therein to expose the top of the well; A plate module including a probe that is fastened to the upper part of the guide module, extends in the depth direction of the receiving portion, and has an oxygen-sensitive material attached to its lower portion; and A system comprising: an oxygen consumption rate measuring module that is fastened to the upper part of the plate module and includes a light-emitting part that transmits light to the oxygen-sensitive material and a sensor part that receives an optical signal generated from the oxygen-sensitive material.

12. In Claim 11, The above impedance measuring unit and the above local field potential measuring unit are electrically connected to the common electrode and the above working electrode, and A system in which the impedance measuring unit applies an alternating current signal to the common electrode to measure the impedance between the common electrode and the working electrode, and the local field potential measuring unit measures the local field potential between the common electrode and the working electrode.

13. In Claim 11, The above probe is located inside the receiving portion of the well, in a system.

14. In Claim 13, The oxygen consumption rate measuring module further includes a driving member that moves the plate module up and down, and A system in which a change in the distance between the probe and the bottom of the well's receiving portion occurs according to the lifting and lowering movement of the plate module.

15. In Claim 11, The above system further includes a processor electrically connected to the cell electrical signal measurement module and the oxygen consumption rate measurement module, and The above processor is a system that analyzes the state of a cell 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.

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