Apparatus and method for multimodal analysis of three-dimensional biological model such as organoid

Abstract

Embodiments of the present invention relate to an apparatus and a method for multimodal analysis of a three-dimensional biometric model such as an organoid. An apparatus according to one aspect comprises: a probe unit which probes the three-dimensional biometric model to obtain an electrical signal and applies electrical stimulation to the three-dimensional biometric model; an optical unit which optically observes the three-dimensional biometric model and applies optical stimulation to the three-dimensional biometric model; and a control unit which controls the probe unit and the optical unit, wherein the probe unit includes a probe extending in a certain direction, and the probe may include a plurality of electrodes arranged along the direction.

Description

Multimodal analysis device and method for three-dimensional biomodels such as organoids

[0001] Embodiments of the present invention relate to a multimodal analysis device and method for a three-dimensional biomodel such as an organoid.

[0002] Organoids are representative biological models of microphysiological systems (MPS), which mimic the cell-cell interactions and biological microenvironment of tissues or organs in vitro. They are widely studied as efficient and accurate tools for drug development, as well as for diagnostic research and regulatory reform. However, despite decades of advancements in biomimetic techniques, standardization and reproducibility remain challenging.

[0003] Meanwhile, the existing 2D multiple electrode array (MEA) is targeted at analyzing biomodels such as neural cell networks cultured in a single layer (2D), but has structural limitations such as extremely limited contact area, which limits the analysis of multilayer (3D) cultured models. Due to these limitations, despite showing high numerical specifications, there is a problem that the analysis utility for 3D biomodels is low. For example, there is a problem that there is a problem in observing signals appearing deep inside, and there is a problem in evaluating electrical characteristics such as potential difference and impedance measurement inside and outside the biomodel. These problems are not significantly improved even when the electrodes are arranged on a curved surface according to the shape of the 3D biomodel.

[0004] 3D biomodels are expensive research materials that require delicate management during a long culture period of approximately several months, and are small in size, less than 1 mm, and easily damaged. Therefore, when performing sensitive analyses on the microenvironment of biomodels, more precise sample manipulation and appropriate control methods are required compared to general experimental environments. However, existing analysis devices or methods have the problem of making it difficult to secure the precision of the inspection process due to excessive reliance on manual operation by the user.

[0005] Furthermore, because complex analysis of biomodels and culture environments is impossible, the performance of interaction studies is extremely limited, and it is difficult to provide multidimensional information on whether the 3D biomodel, which is the subject of analysis, has sufficient functional characteristics.

[0006] According to one aspect of the present invention, a device and method for analyzing a three-dimensional biomodel, which can precisely control the analysis process of a three-dimensional biomodel such as an organoid in real time, can be provided.

[0007] According to another aspect of the present invention, a device and method for analyzing a 3D biomodel can be provided, which can perform real-time integrated analysis and feedback on the function of a 3D biomodel, thereby enabling quality management and evaluation of a biomodel to be analyzed prior to bioanalysis using the 3D biomodel.

[0008] According to another aspect of the present invention, a device and method for analyzing a three-dimensional biomodel can be provided, which enable various analyses of a three-dimensional biomodel in a non-destructive manner.

[0009] According to another aspect of the present invention, a device and method for analyzing a three-dimensional biomodel can be provided, which can perform various functional analyses of a three-dimensional biomodel with a single device.

[0010] According to one embodiment of the present invention, a device is an analysis device for a three-dimensional biomodel, comprising: a probe unit for probing the three-dimensional biomodel to obtain an electrical signal and apply an electrical stimulus to the three-dimensional biomodel; an optical unit for optically observing the three-dimensional biomodel and applying an optical stimulus to the three-dimensional biomodel; and a control unit for controlling the probe unit and the optical unit; wherein the probe unit includes a probe extending in one direction, and the probe may include a plurality of electrodes arranged along the direction.

[0011] In one embodiment, the device may include at least one selected from the group consisting of cardiac organoids, retinal organoids, brain organoids, tumor organoids, neural organoids, and organoids comprising cells subjected to optogenetic manipulation.

[0012] In a device according to one embodiment, the probe may be provided in multiple numbers.

[0013] In a device according to one embodiment, the probe may further include one or more ion-selective membranes covering the surfaces of at least some of the plurality of electrodes.

[0014] In a device according to one embodiment, the plurality of electrodes are classified into a first group arranged along the direction on one side of the probe and a second group arranged along the direction on the other side of the probe, and one of the first group and the second group may have a surface covered by the one or more ion-selective membranes.

[0015] In a device according to one embodiment, the probe unit may further include a driving unit that controls the position of the probe.

[0016] In a device according to one embodiment, the probe unit may further include a measuring unit that measures electrical signals obtained from at least some of the plurality of electrodes of the probe.

[0017] In a device according to one embodiment, the measurement unit can measure electrical signals according to depth within the three-dimensional biomodel.

[0018] In a device according to one embodiment, the electrical signal may include at least one of a biopotential and a bioimpedance of the three-dimensional biomodel.

[0019] In a device according to one embodiment, the probe unit may further include an electrical stimulation unit that applies electrical stimulation to the three-dimensional biomodel via the probe.

[0020] In one embodiment of the device, the electrical stimulation may be a current pulse or voltage pulse in the form of a pulse train or a single pulse.

[0021] In a device according to one embodiment, the optical unit may include an observation unit that acquires optical information of the three-dimensional biomodel in real time.

[0022] In a device according to one embodiment, the optical unit may include a light stimulation unit that applies light stimulation to the three-dimensional biomodel.

[0023] In a device according to one embodiment, the optical stimulus may be pulsed light.

[0024] In a device according to one embodiment, the control unit may include a signal processing unit that processes information transmitted from the probe unit and an image processing unit that processes information transmitted from the optical unit.

[0025] In a device according to one embodiment, the control unit may include a control unit that controls the operation of the probe unit and the optical unit based on information processed from at least one of the signal processing unit and the image processing unit.

[0026] A method according to one embodiment of the present invention is a method for analyzing a three-dimensional biomodel using a device according to one embodiment of the present invention, the method including: a step of inserting the probe into the three-dimensional biomodel; a step of verifying a process of inserting the probe into the three-dimensional biomodel; a step of obtaining an electrical signal of the three-dimensional biomodel; a step of applying an electrical stimulus or an optical stimulus to the three-dimensional biomodel; and a step of determining whether an electrical signal measured from the three-dimensional biomodel satisfies a preset condition.

[0027] In a method according to one embodiment, the electrical stimulation is a current pulse in the form of a pulse train, and the current pulse can satisfy at least one of the following conditions.

[0028] a. Pulse width is 0.1 ms to 1 ms,

[0029] b. The frequency of the pulse train is 0.1 Hz to 10 Hz,

[0030] c. Peak is 1nA to 1mA.

[0031] In a method according to one embodiment, the optical stimulus is pulsed light, and the pulsed light can satisfy at least one of the following conditions.

[0032] a. Pulse width of 0.01ms to 1,000ms,

[0033] b. Intensity is 0.01 mW / cm 2 1,000mW / cm 2 ,

[0034] c. Wavelength is 300 nm to 800 nm.

[0035] In a method according to one embodiment, the three-dimensional biomodel can be analyzed while maintaining its original shape.

[0036] According to one aspect of the present invention, a device and method for analyzing a three-dimensional biomodel, which can precisely control the analysis process of a three-dimensional biomodel such as an organoid in real time, can be provided.

[0037] According to another aspect of the present invention, a device and method for analyzing a 3D biomodel can be provided, which can perform real-time integrated analysis and feedback on the function of a 3D biomodel, thereby enabling quality management and evaluation of a biomodel to be analyzed prior to bioanalysis using the 3D biomodel.

[0038] According to another aspect of the present invention, a device and method for analyzing a three-dimensional biomodel can be provided, which enable various analyses of a three-dimensional biomodel in a non-destructive manner.

[0039] According to another aspect of the present invention, a device and method for analyzing a three-dimensional biomodel can be provided, which can perform various functional analyses of a three-dimensional biomodel with a single device.

[0040] FIG. 1 is a block diagram functionally illustrating an example of a device according to one embodiment of the present invention.

[0041] Figure 2 is a schematic diagram for structurally explaining an example of a device according to one embodiment of the present invention.

[0042] FIG. 3 is a drawing showing an example of a probe according to one embodiment of the present invention.

[0043] Figure 4 is a drawing for explaining analysis of a three-dimensional biomodel using multiple probes.

[0044] FIG. 5 is a drawing showing an example of a probe according to another embodiment of the present invention.

[0045] FIG. 6 is an image showing a cardiac organoid being analyzed using a device according to one embodiment of the present invention.

[0046] FIG. 7 is an image showing an example of local biopotential profiling of a cardiac organoid obtained using a device according to one embodiment of the present invention.

[0047] FIG. 8 is an image showing a cardiac organoid being analyzed using a device according to another embodiment of the present invention.

[0048] FIG. 9 is an image showing an example of bioimpedance profiling of a cardiac organoid obtained using a device according to another embodiment of the present invention.

[0049] FIG. 10 is a schematic diagram schematically showing an analysis of a retinal organoid using a device according to another embodiment of the present invention.

[0050] FIG. 11 is an image showing an example of in vivo spike profiling of a retinal organoid obtained using a device according to another embodiment of the present invention.

[0051] Figure 12 is a schematic diagram schematically showing an analysis of the bio-ion concentration of an organoid using a device according to another embodiment of the present invention.

[0052] Figure 13 is a step diagram showing an example of a method for analyzing a three-dimensional biomodel according to one embodiment of the present invention.

[0053] Figure 14 is a graph showing an example of stabilizing a cardiac organoid by applying electrical stimulation.

[0054] Figure 15 is a graph showing the biopotential profiles 3 minutes (A), 4 minutes (B), 5 minutes (C), and 6 minutes (D) after inserting the probe into the cardiac organoid.

[0055] The embodiments described herein may be modified in many different forms, and thus the technology according to one implementation is not limited to the embodiments described below. Furthermore, throughout the specification, the terms "comprises," "includes," "contains," or "has" a component do not exclude other components unless specifically stated to the contrary, but rather imply that other components may be included, and do not exclude additional elements, materials, or processes that are not listed.

[0056] As used herein, "identical" or "uniform" may mean identical or uniform within an acceptable margin of error, unless otherwise specified. For example, "identical" in certain components or material property measurements may mean that the two compared objects are not only completely identical but also identical within a margin of error. Meanwhile, "identical" in certain material property measurements may mean that the difference in the measured values ​​between objects is approximately less than 5%, specifically less than 3%, or more specifically less than 1%.

[0057] In this specification, the angle formed by two objects being perpendicular, or parallel or parallel to each other may include not only being geometrically perpendicular or parallel, but also being within a slight margin of error.

[0058] A numerical range as used herein includes a lower bound and an upper bound and all values ​​within that range, increments logically derived from the shape and width of the defined range, all doubly delimited values, and all possible combinations of upper and lower bounds of numerical ranges delimited in different shapes.

[0059] Unless otherwise specified herein, “about” may be considered a value within 30%, 25%, 20%, 15%, 10% or 5% of the stated value.

[0060] The use of terms such as "first," "second," and "third" in front of certain components in this specification is solely to avoid confusion regarding the components they refer to, and is unrelated to the order, importance, or dominant-subordinate relationship between the components. For example, an invention can be implemented that includes only a second component without a first component.

[0061] In this specification, the term "electrically connected" may mean, without limitation, any connection method by which a plurality of objects can be connected to each other so that they can electrically communicate with each other, and may be implemented in various ways, such as by directly connecting the plurality of objects to each other, or by connecting them via a third object.

[0062] In this specification, a component defined as “…part” or “…unit” may mean, without limitation, a single component or a set of two or more identical or similar components that have something in common in a functional aspect, and the set of components may be configured by a non-limiting combination of hardware and / or software.

[0063] The present invention will be described in detail below. However, the following description is merely exemplary, and the present invention is not limited to the embodiments described below.

[0064] FIG. 1 is a block diagram functionally illustrating an example of a device according to one embodiment of the present invention.

[0065] Figure 2 is a schematic diagram for structurally explaining an example of a device according to one embodiment of the present invention.

[0066] FIG. 3 is a drawing showing an example of a probe according to one embodiment of the present invention.

[0067] Referring to FIGS. 1 to 3, a device (10) according to one aspect of the present invention is an analysis device (10) for a three-dimensional biomodel (50), comprising: a probe unit (100) for probing the three-dimensional biomodel (50) to obtain an electrical signal and apply an electrical stimulus to the three-dimensional biomodel (50); an optical unit (200) for optically observing the three-dimensional biomodel (50) and applying an optical stimulus to the three-dimensional biomodel (50); and a control unit (300) for controlling the probe unit (100) and the optical unit (200). The probe unit (100) includes a probe (110) extending in one direction, and the probe (110) may include a plurality of electrodes (111) arranged along the direction (DRE).

[0068] In one embodiment, the device (10) may be a device for analyzing the three-dimensional biomodel (50). In an exemplary embodiment, the device (10) may be a device for analyzing the three-dimensional biomodel (50) electrically or electrochemically.

[0069] In one embodiment, the three-dimensional biomodel (50) may be multiple. This means that the device (10) can analyze multiple three-dimensional biomodels (50) simultaneously.

[0070] In an exemplary embodiment, the three-dimensional biomodel (50) may be a tissue isolated from a living organism or a cultured cell culture. For example, the cell culture may be a spheroid or an organoid.

[0071] In one embodiment, the three-dimensional biomodel (50) may include a biomodel capable of generating an electrical signal or a biomodel capable of generating a signal in response to a light stimulus.

[0072] In one embodiment, the three-dimensional biomodel (50) may include at least one selected from the group consisting of cardiac organoids, retinal organoids, brain organoids, tumor organoids, neural organoids, and organoids comprising cells subjected to optogenetic manipulation.

[0073] In one embodiment, the three-dimensional biomodel (50) may be a cardiac organoid (CO) or a retinal organoid (RO). Although not necessarily limited thereto, the device (10) according to one aspect of the present invention may disclose a configuration optimized for multimodal analysis of organoids. This will be described later.

[0074] Referring again to FIGS. 1 and 2, in one embodiment, the probe unit (100) can probe the three-dimensional biomodel (50) to measure an electrical signal or apply an electrical stimulus to the three-dimensional biomodel (50).

[0075] To this end, in one embodiment, the probe unit (100) includes a probe (110), and the probe (110) may extend in one direction (DRE) and include a plurality of electrodes (111) arranged along the direction (DRE).

[0076] In one embodiment, the probe (110) may come into contact with the three-dimensional biomodel (50) during the analysis process of the three-dimensional biomodel (50). In an exemplary embodiment, the probe (110) may be invasively inserted into the three-dimensional biomodel (50). Accordingly, at least a portion of the probe (110) may enter the interior of the three-dimensional biomodel (50), and may measure electrical signals generated within the three-dimensional biomodel (50), or may apply electrical stimulation to the interior of the three-dimensional biomodel (50).

[0077] Referring to FIGS. 2 and 3, in one embodiment, the probe (110) may extend in one direction (DRE). In the present specification, the direction (DRE) merely refers to the direction (DRE) in which the probe (110) extends, and the absolute direction within the device (10) is not particularly limited. For example, the direction (DRE) may be inclined with respect to the height direction in the three-dimensional spatial coordinate system as illustrated in FIG. 2, but is not necessarily limited thereto, and may be parallel to or perpendicular to the height direction as needed.

[0078] Meanwhile, referring to FIGS. 2 and 3, the probe (110) may have a width at the end smaller than the width of the remaining portion in order to be inserted more smoothly into the three-dimensional biomodel (50), and at least a portion of the end may have a pointed shape. However, the present invention is not necessarily limited thereto, and if necessary, at least a portion of the end may have a blunt shape, or the width of the end and the width of the remaining portion may be substantially the same. Meanwhile, the end may mean one end of the probe (110) that is intended to be the first contact point with the three-dimensional biomodel (50).

[0079] Meanwhile, in one embodiment, the probe (110) can enter the three-dimensional biomodel (50) along the direction in which the probe (110) extends (DRE). As will be described later, information according to the depth within the three-dimensional biomodel (50) can be obtained by the probe (110) having the above-described shape.

[0080] In one embodiment, the probe (110) may comprise a chemically stable material that does not electrochemically affect the three-dimensional biomodel (50). For example, the material may be silicon. In this embodiment, the probe (110) may be a probe made of silicon.

[0081] In one embodiment, the probe (110) may include a plurality of electrodes (111) arranged along the direction (DRE). In such an embodiment, the probe (110) includes a plurality of electrodes (111), and each of the plurality of electrodes (111) may be arranged along the direction (DRE). Therefore, as will be described later, based on the electrical signals obtained from each of the plurality of electrodes (111), electrical signals according to the depth within the three-dimensional biomodel (50) may be measured. The electrode may be, for example, an electrode containing platinum, but is not necessarily limited thereto.

[0082] Meanwhile, the configurations illustrated in FIGS. 2 and 3 are provided to explain the configuration of the device (10) of the present invention, and the present invention is not limited to what is illustrated in FIGS. 2 and 3. For example, the number, position, size, ratio, shape, shading, etc. of each configuration illustrated in FIGS. 2 and 3 are arbitrary and can be freely configured within a range that does not violate the matters defined in the present invention. This can also be applied to all of the drawings below.

[0083] Figure 4 is a drawing for explaining analysis of a three-dimensional biomodel using multiple probes.

[0084] Referring to FIG. 4, in one embodiment, the probe (110) may be provided in multiple numbers.

[0085] In one embodiment, all of the probes (110) provided in a plurality may be used for analysis of the three-dimensional biomodel (50). In an exemplary embodiment, the probes (110) may be provided in two numbers. The two probes (110) may each be inserted into the three-dimensional biomodel (50). Meanwhile, in FIG. 4, internal shading is illustrated in the three-dimensional biomodel (50), but this is only to indicate that the three-dimensional biomodel (50) has depth, and is not intended to specify or limit the shape of the three-dimensional biomodel (50).

[0086] At this time, as illustrated in FIG. 4, the direction (DRE) in which each of the probes (110) extends may have the same absolute direction in the three-dimensional space coordinate system. However, it is not necessarily limited thereto, and as illustrated in FIG. 6, which will be described later, the direction (DRE) in which each of the probes (110) extends may have different absolute directions in the three-dimensional space coordinate system. In this case, as illustrated in FIG. 6, three probes (110) are provided, and two of the probes (110) may be positioned adjacent to each other and arranged so that their absolute directions are the same in the three-dimensional space coordinate system, and the remaining one probe (110) may be positioned spaced apart from the two probes (110) and arranged so that their absolute directions are different.

[0087] Referring to FIG. 4, in one embodiment, the spacing (d) between the plurality of probes (110) may be flexibly set according to the size of the three-dimensional biomodel (50). For example, the spacing (d) may be 10 μm to 1,000 μm. In a specific embodiment, the spacing (d) may be 50 μm or more, 100 μm or more, or 150 μm or more, or 900 μm or less, 800 μm or less, 700 μm or less, 600 μm or less, 500 μm or less, or 400 μm or less. For example, the spacing (d) may be 250 μm.

[0088] Meanwhile, since all of the probes (110) provided in multiple units as described above are used for the analysis of the three-dimensional biomodel (50), it is possible to more precisely verify whether the probes (110) have properly entered the three-dimensional biomodel (50), or to more precisely analyze the internal structure, state, etc. of the three-dimensional biomodel (50).

[0089] Meanwhile, according to an exemplary embodiment, the probe (110) as described above may utilize a commercial probe. For example, H2 from Cambridge Neurotech may be used, but is not necessarily limited thereto.

[0090] FIG. 5 is a drawing showing an example of a probe according to another embodiment of the present invention.

[0091] FIG. 5 illustrates an example of a probe (110') according to another embodiment of the present invention. Referring to FIG. 5, in one embodiment, the probe (110') may further include one or more ion-selective membranes (112) covering the surfaces of at least some of the electrodes (111) among the plurality of electrodes (111).

[0092] In one embodiment, the ion-selective membrane (112) may refer to any membrane capable of selectively permeating ions without limitation. In an exemplary embodiment, the ion-selective membrane may be, but is not necessarily limited to, PEDOT:PSS, a sodium ionophore, a potassium ionophore, or a calcium ionophore.

[0093] In one embodiment, at least some of the surfaces of the plurality of electrodes (111) positioned on the probe (110') may be covered by the ion-selective membrane (112). In an exemplary embodiment, the surfaces may be overlapped with the ion-selective membrane (112) for electrochemical stability. For example, the surfaces may be covered with PEDOT:PSS, and the surface of the PEDOT:PSS may be additionally covered with an ionophore.

[0094] In one embodiment, electrodes (111) other than those whose surfaces are covered by the ion-selective membrane (112) may not have their surfaces covered by the ion-selective membrane (112).

[0095] Referring again to FIG. 5, in one embodiment, the plurality of electrodes (111) are classified into a first group (111A) arranged along the direction (DRE) on one side of the probe (110') and a second group (111B) arranged along the direction (DRE) on the other side of the probe (110'), and either group of the first group (111A) or the second group (111B) may have its surface covered by one or more ion-selective membranes (112).

[0096] In one embodiment, the plurality of electrodes (111) may be classified into a first group (111A) and a second group (111B). Each of the first group (111A) and the second group (111B) is composed of a plurality of electrodes (111), and each of the plurality of electrodes (111) constituting the first group (111A) may be arranged along the direction (DRE), and each of the plurality of electrodes (111) constituting the second group (111B) may be arranged along the direction (DRE).

[0097] In one embodiment, the number of the plurality of electrodes (111) constituting the first group (111A) and the number of the plurality of electrodes (111) constituting the second group (111B) may be the same. In one embodiment, the spacing between the plurality of electrodes (111) constituting the first group (111A) and the spacing between the plurality of electrodes (111) constituting the second group (111B) may be the same.

[0098] In one embodiment, the first group (111A) and the second group (111B) may be symmetrical along any one of the planes including the direction (DRE).

[0099] In the above embodiment, either one of the first group (111A) and the second group (111B) may have its surface covered by one or more ion-selective membranes (112). Accordingly, the manufacturing processability or usability of the probe (110') may be further enhanced. Meanwhile, although FIG. 5 illustrates an embodiment in which the electrodes (111) of the first group (111A) have their surfaces covered by the ion-selective membranes (112), it should be understood that the present invention is not necessarily limited thereto.

[0100] Meanwhile, in an exemplary embodiment, the ion selective membrane (112) may be coated on the surface of the electrode (111) to cover the surface of the electrode (111), but is not necessarily limited thereto.

[0101] In one embodiment, the ion selective membrane (112) may be a single membrane.

[0102] In one embodiment, the ion selective membrane (112) may be provided in multiple numbers.

[0103] In one embodiment, the ion-selective membrane (112) is provided in multiple numbers, and all of the ion-selective membranes (112) can have selectivity for the same ion. That is, all of the ion-selective membranes (112) can have selective permeability for the same ion.

[0104] As will be described later, the probe (110') of the above embodiment can measure the concentration of a specific ion according to the depth of the three-dimensional biomodel (50).

[0105] In one embodiment, the ion selective membrane (112) is provided in a plurality of pieces, and at least two of the plurality of ion selective membranes (112) can have selectivity for different ions.

[0106] That is, some of the plurality of ion-selective membranes (112) may have selective permeability to one ion, and other of the ion-selective membranes (112) may have selective permeability to another ion. Of course, other of the ion-selective membranes (112) may have selective permeability to another ion.

[0107] By using the probe (110') of the above embodiment, the concentration of a specific ion inside an arbitrary three-dimensional biomodel (50) can be estimated or measured with high accuracy, and the measurement can be performed more simply. In the case of the conventional technology, in order to precisely measure the concentration of a specific ion inside a three-dimensional biomodel (50), there was a difficulty in that the potential change at all interfaces of the three-dimensional biomodel (50) had to be known. However, by using the probe (110') of the above embodiment, the concentration of a specific ion inside an arbitrary three-dimensional biomodel (50) can be estimated or measured with high accuracy even without the above information. The details of this will be described later.

[0108] Referring again to FIGS. 1 and 2, in one embodiment, the probe unit (100) may further include a driving unit (120) that controls the position of the probe (110).

[0109] In one embodiment, the drive unit (120) can control the position of the probe (110). In an exemplary embodiment, the drive unit (120) can control the position of the probe (110) macroscopically or microscopically. Meanwhile, in one embodiment, one end of the probe (110) can be in contact with the drive unit (120). That is, one end of the probe (110) can be in contact with the drive unit (120), and the other end can be in contact with the three-dimensional biomodel (50).

[0110] In one embodiment, the driving unit (120) can control the position of the probe (110) on a macroscopic level. For example, before analysis using the device (10) begins, the probe (110) can be positioned apart from the 3D biomodel (50) to be analyzed. When analysis using the device (10) begins, the probe (110) can be positioned adjacent to the 3D biomodel (50) to be analyzed, or moved to come into contact with the 3D biomodel (50). The driving unit (120) can control the position of the probe (110) by moving it as described above.

[0111] In one embodiment, the driving unit (120) can microscopically control the position of the probe (110). For example, the probe (110) can be controlled to enter the inside of the three-dimensional biomodel (50) while positioning it adjacent to the three-dimensional biomodel (50) or in contact with the three-dimensional biomodel (50). In order to precisely perform analysis on a three-dimensional biomodel (50) having a generally microscopic size, the movement distance of the probe (110) can be microscopically controlled while it has entered the inside of the three-dimensional biomodel (50). For example, the movement as described above can be microscopically controlled along the direction (DRE) in which the probe (110) extends.

[0112] In one embodiment, the drive unit (120) can also control the angle of the probe (110) macroscopically or microscopically. Here, the angle of the probe (110) may mean an absolute direction within the spatial coordinate system in the direction (DRE) in which the probe (110) extends, as described above.

[0113] In one embodiment, the driving unit (120) may be operated by a user operating a separately provided position driving means (not shown), and, if necessary, the operation of the driving unit (120) may be automatically performed by the device (10).

[0114] In one embodiment, unlike that illustrated in FIG. 2, the driving unit (120) may be formed in multiple units. In this case, each driving unit (120) may have one or more probes (110) independently positioned.

[0115] Referring again to FIG. 2, the device (10) may further include a fixing member (400) for fixing the three-dimensional biomodel (50).

[0116] In one embodiment, the fixing member (400) can fix one or more three-dimensional biomodels (50).

[0117] The above-mentioned fixing member (400) may include a microwell, but is not necessarily limited thereto, and any means that can fix the three-dimensional biomodel (50) to maintain its original shape while remaining in the same spatial position even when a certain amount of external force is applied may be employed without limitation.

[0118] In one embodiment, the fixing member (400) may further include a temperature maintenance device (not shown). In one embodiment, the temperature maintenance device (not shown) may employ any known device without limitation as long as it corresponds to a device capable of maintaining a constant temperature in a specific space. The temperature maintenance device (not shown) may be provided to maintain homeostasis of the three-dimensional biomodel (50) fixed to the fixing member (400).

[0119] In one embodiment, the fixing member (400) may further include an XYZ stage (not shown). The XYZ stage (not shown) is provided at the lower end of the fixing member (400) and can three-dimensionally control the position of the fixing member (400). The operation of the XYZ stage (not shown) may be performed by a user's manipulation, or may be automatically performed by the device (10) if necessary.

[0120] Referring again to FIG. 1, in one embodiment, the probe unit (100) may further include a measuring unit (130) that measures electrical signals obtained from at least some of the electrodes (111) among the plurality of electrodes (111) of the probe (110).

[0121] In an exemplary embodiment, the measurement unit (130) can measure electrical signals obtained from the plurality of electrodes (111) in real time.

[0122] When the probe (110) enters the inside of the three-dimensional biomodel (50), at least some of the electrodes (111) among the plurality of electrodes (111) may be located inside the three-dimensional biomodel (50). Meanwhile, the remaining electrodes (111) that are not located inside the three-dimensional biomodel (50) may be located outside the three-dimensional biomodel (50).

[0123] Meanwhile, each of the plurality of electrodes (111) can obtain an electrical signal. At this time, the electrical signal obtained from each of the plurality of electrodes (111) can be transmitted to the measuring unit (130). Accordingly, the measuring unit (130) can measure the electrical signal obtained from each of the plurality of electrodes (111) and obtain information related thereto.

[0124] In one embodiment, the measurement unit (130) can measure electrical signals according to depth within the three-dimensional biomodel (50).

[0125] As described above, the probe (110) may include a plurality of electrodes (111) arranged along the direction (DRE), and thus, an electrical signal may be obtained from each of the plurality of electrodes (111). When the probe (110) enters the inside of the three-dimensional biomodel (50), at least some of the electrodes (111) among the plurality of electrodes (111) may be located inside the three-dimensional biomodel (50).

[0126] Accordingly, based on the electrical signals obtained from the electrodes (111) located inside the 3D biomodel (50), electrical signals according to the depth within the 3D biomodel (50) can be measured.

[0127] Meanwhile, although it may vary depending on the analysis conditions, at least some of the electrodes (111) that are not located inside the 3D biomodel (50) may also measure electrical signals. In this case, the electrodes (111) that are not located inside the 3D biomodel (50) and from which electrical signals are measured may be in contact with media or the like. In this aspect, some of the plurality of electrodes (111) are located inside the 3D biomodel (50) and thus an electrical signal can be obtained according to the depth within the 3D biomodel (50), and at least some of the remaining electrodes (111) that are not located inside the 3D biomodel (50) among the plurality of electrodes (111) may obtain an electrical signal by coming into contact with the media or the like. Accordingly, the electrical signal obtained by coming into contact with the media or the like serves as a kind of reference, and based on this, the electrical signal obtained within the 3D biomodel (50) can be verified.

[0128] In one embodiment, the electrical signal measured by the measurement unit (130) may be transmitted to and processed by the signal processing unit (310) of the control unit (300), which will be described later. The processed information may be displayed on a display unit (not shown) to be described later and provided to a user, or transmitted to and stored in a storage unit (not shown) to be described later, thereby being utilized as analysis data for a three-dimensional biomodel (50), or provided to a control unit (330) to be described later and function as information that serves as a basis for judgment for control in the control unit (330).

[0129] In one embodiment, the electrical signal may include at least one of a biopotential and a bioimpedance of the three-dimensional biomodel (50).

[0130] In one embodiment, the electrical signal may be a biopotential of the three-dimensional biomodel (50).

[0131] In one embodiment, the electrical signal may be the bioimpedance of the three-dimensional biomodel (50).

[0132] In addition, detailed information about the above electrical signals and the bio-information of the three-dimensional bio-model (50) that can be obtained based on the electrical signals will be described later.

[0133] Referring again to FIG. 1, in one embodiment, the probe unit (100) may further include an electrical stimulation unit (140) that applies electrical stimulation to the three-dimensional biomodel (50) via the probe (110).

[0134] In one embodiment, the electrical stimulation unit (140) can apply an electrical stimulus to the three-dimensional biomodel (50) via the probe (110). In an exemplary embodiment, the electrical stimulation unit (140) is electrically connected to a separately provided power supply (not shown) to provide current or voltage to at least some of the electrodes (111) formed on the probe (110), thereby applying an electrical stimulus to the three-dimensional biomodel (50).

[0135] In an exemplary embodiment, the three-dimensional biomodel (50) to which electrical stimulation is applied by the electrical stimulation unit (140) may be a cardiac organoid.

[0136] In an exemplary embodiment, the electrical stimulation may be applied to establish quality control of the three-dimensional biomodel (50) being analyzed. For example, an electrical pacing stimulus may be applied to the three-dimensional biomodel (50) via the probe (110).

[0137] When cultured cardiac organoids are removed and transferred to the device (10) for analysis, the cardiac organoids may exhibit abnormal behavior or conditions, such as irregular beating or failing to reach a normal heart rate, due to stress caused by environmental changes. Therefore, in order to perform precise and accurate analysis using the cardiac organoids, the condition of the cardiac organoids must be stabilized in advance. For such stabilization, electrical stimulation can be applied from the electrical stimulation unit (140) to the three-dimensional biomodel (50), for example, the cardiac organoid.

[0138] As described above, in one embodiment, the three-dimensional biomodel (50) can be stabilized by the electrical stimulation. In an exemplary embodiment, the cardiac organoid can be stabilized by the electrical stimulation.

[0139] That is, the three-dimensional biomodel (50), for example, the cardiac organoid, can be stabilized by the electrical stimulation applied from the electrical stimulation unit (140). Details thereof will be described later.

[0140] In one embodiment, the electrical stimulation may be a current pulse or voltage pulse in the form of a pulse train or a single pulse. Other details are described below.

[0141] Referring again to FIGS. 1 and 2, in one embodiment, the optical unit (200) can optically observe the three-dimensional biomodel (50) or apply an optical stimulus to the three-dimensional biomodel (50).

[0142] Referring to FIGS. 1 and 2, in one embodiment, the optical unit (200) may include an observation unit (210) that acquires optical information of the three-dimensional biomodel (50) in real time.

[0143] In one embodiment, the observation unit (210) can acquire optical information of the three-dimensional biomodel (50) in real time. In an exemplary embodiment, the optical information may include an image or video of the three-dimensional biomodel (50). In an exemplary embodiment, the optical information may mean a two-dimensional image or video of the three-dimensional biomodel (50).

[0144] For this purpose, the observation unit (210) may employ any known means for obtaining real-time images or videos without limitation.

[0145] In one embodiment, the optical information acquired from the observation unit (210) may be transmitted to and processed by the image processing unit (320) of the control unit (300), which will be described later. The processed information may be displayed on a display unit (not shown) to be described later and provided to a user, or transmitted to and stored in a storage unit (not shown) to be described later, thereby being utilized as analysis data for a three-dimensional biomodel (50), or further provided to a control unit (330) to be described later and function as information that serves as a basis for judgment for control in the control unit (330).

[0146] Referring again to FIGS. 1 and 2, in one embodiment, the optical unit (200) may include a light stimulation unit (220) that applies a light stimulus (L) to the three-dimensional biomodel (50).

[0147] In one embodiment, the optical stimulation unit (220) can apply optical stimulation (L) to the three-dimensional biomodel (50). For this purpose, in an exemplary embodiment, the optical stimulation unit (220) can be equipped with a light source.

[0148] In one embodiment, the light source may include, but is not necessarily limited to, a multi-wavelength LED or laser irradiation device. The light source may not be particularly limited as long as it corresponds to a device capable of irradiating light (e.g., continuous light or pulsed light) with desired characteristics to a desired target.

[0149] In an exemplary embodiment, the three-dimensional biomodel (50) to which the optical stimulus (L) is applied by the optical stimulus unit (220) may be a retinal organoid.

[0150] In an exemplary embodiment, the optical stimulus (L) may be applied to establish quality control of a three-dimensional biomodel (50) to be analyzed. For example, by analyzing the feedback of the three-dimensional biomodel (50) by the optical stimulus (L), it is possible to determine whether the three-dimensional biomodel (50) is in a stable state and / or normal state.

[0151] In the case of retinal organoids, they can react to light applied from the outside, and can react in proportion to the intensity of the applied light. The reaction of the retinal organoids to the light stimulus (L) applied by the light stimulus unit (220) can be observed in the form of an electrical signal, which can be measured by the measurement unit (130) through the inserted probe (110).

[0152] Based on the information measured by the measurement unit (130), the status of the retinal organoid can be determined. If a reaction satisfying a preset condition is shown, it can be indicated that the status of the retinal organoid being analyzed is normal and / or stable.

[0153] Referring again to FIGS. 1 and 2, in one embodiment, the control unit (300) can control the probe unit (100) and the optical unit (200).

[0154] In one embodiment, the control unit (300) may process information transmitted from the probe unit (100) and the optical unit (200), and control the operation of the probe unit (100) and the optical unit (200) based on the processed information. Meanwhile, the information processed as described above may be displayed on a display unit (not shown) to be described later and provided to a user, or may be transmitted to and stored in a storage unit (not shown) to be described later, thereby being utilized as analysis data for a three-dimensional biomodel (50).

[0155] In one embodiment, the control unit (300) may include a signal processing unit (310) that processes information transmitted from the probe unit (100).

[0156] As in the above-described embodiment, the measuring unit (130) of the probe unit (100) can measure electrical signals obtained from at least some of the electrodes (111) among the plurality of electrodes (111), and thus can obtain electrical signals within the three-dimensional biomodel (50).

[0157] The electrical signal within the three-dimensional biomodel (50) obtained from the measuring unit (130) may be transmitted to the signal processing unit (310) and processed. The signal processing unit (310) may filter or plot in real time the electrical signal (biopotential or bioimpedance) transmitted from the measuring unit (130). Through such processing, the signal processing unit (310) may analyze the electrical bioinformation within the three-dimensional biomodel (50) in real time.

[0158] Based on the electrical bio-information analyzed in real time in this way, the local field potential, bio-impedance, bio-spike, and / or bio-ion concentration within the 3D bio-model (50) can be profiled in real time.

[0159] FIG. 6 is an image showing a cardiac organoid being analyzed using a device according to one embodiment of the present invention.

[0160] FIG. 7 is an image showing an example of local biopotential profiling of a cardiac organoid obtained using a device according to one embodiment of the present invention.

[0161] Hereinafter, in order to explain in detail the embodiments of FIGS. 6 and 7, the probes are illustrated with separate drawing reference numerals (110-1, 110-2).

[0162] The above three-dimensional biomodel (50) may contain cells within it. At least some of the cells may emit electrophysiological signals. Accordingly, local biopotentials at various depths and locations within the three-dimensional biomodel (50) may change in real time.

[0163] Meanwhile, cardiac organoids can have a unique waveform according to the heartbeat. Therefore, by spatiotemporally analyzing the waveform measured in this way using the device according to an embodiment of the present invention, information about the location of specific cells within the cardiac organoid, conduction velocity, and wave propagation direction can be obtained. Furthermore, it is possible to non-destructively determine whether the cardiac organoid is in a normal or abnormal state, or to verify whether the probe (110-1, 110-2) is correctly positioned within the cardiac organoid, inserted to a certain extent, or inserted to an appropriate degree.

[0164] FIG. 6 illustrates an example of analyzing a cardiac organoid (CO) using the device of the present invention. Specifically, it illustrates two probes (110-1, 110-2) being inserted into the cardiac organoid (CO) to measure biopotential.

[0165] The biopotential measured as described above can be transmitted to the signal processing unit via the measurement unit. The signal processing unit can process the transmitted biopotential-related signal to profile the local biopotential. An example of data actually acquired according to this method is shown in Fig. 7.

[0166] The graphs on the right side of FIG. 7 are profiling of potentials over time measured from the probe (110-1) located closer to the center of the cardiac organoid (CO) among the two probes (110-1) shown in FIG. 6, and the graphs on the left side are profiling of potentials over time measured from the probe (110-2) located further from the center of the cardiac organoid (CO). Meanwhile, the graphs illustrated in FIG. 7 are profiling of local biopotentials measured from electrodes located along the extension direction of the probes (110-1, 110-2) from the ends of the probes (110-1, 110-2) from the bottom to the top (i.e., located along the direction toward the drive unit (120) described with reference to FIGS. 1 and 2).

[0167] In both the graphs on the right and left of Figure 7, it can be confirmed that potentials with a waveform having a constant period are observed from the bottom to a specific location upward. Based on the observation of potentials with a waveform having a constant period, it can be assumed that the electrodes where the potentials are observed are located within the organoid. Meanwhile, each waveform can differ depending on the depth, and as observed in Figure 7, it can be confirmed that potentials with different waveforms are observed depending on the location (depth) of the electrode.

[0168] The local biopotential profiled as described above can be acquired according to the depth of a three-dimensional biomodel (50), for example, a cardiac organoid (CO). With reference to this, the location, conduction velocity, and wave propagation direction of specific cells within the organoid can be analyzed, and whether the organoid condition is normal or abnormal can be determined, or whether the probe (110-1, 110-2) is correctly positioned within the organoid, how deep it is inserted, or whether it is inserted to an appropriate degree can be verified in real time.

[0169] FIG. 8 is an image showing a cardiac organoid being analyzed using a device according to another embodiment of the present invention.

[0170] FIG. 9 is an image showing an example of bioimpedance profiling of a cardiac organoid obtained using a device according to another embodiment of the present invention.

[0171] Hereinafter, in order to explain in detail the embodiments of FIGS. 8 and 9, the probes are illustrated with separate drawing reference numerals (110-1', 110-2').

[0172] The above 3D biomodel may contain cells within it, and thus its internal structure may be complex. Depending on the individual state of the cells and the internal structure, the 3D biomodel may exhibit varying impedances at different locations.

[0173] In general, locations with dense structures, such as connective tissue, can exhibit high bioimpedance. Conversely, locations with internal voids or cytoplasm can exhibit low bioimpedance. Furthermore, increased cell death or edema can increase conductivity, leading to a decrease in impedance. Furthermore, as cells proliferate or tissues mature, the extracellular matrix (ECM) forms at a high density, which can lead to an increase in impedance.

[0174] Accordingly, by spatiotemporally analyzing the bio-impedance measured in this way using the device according to the embodiment of the present invention, it is possible to non-destructively determine whether the state of the three-dimensional bio-model is normal or abnormal, and also to determine or track the internal structure, compartment, and bio-state of the three-dimensional bio-model, or to verify whether the probe (110-1', 110-2') is correctly positioned within the three-dimensional bio-model (50), or to what extent it is inserted, or to an appropriate extent.

[0175] FIG. 8 illustrates an example of analyzing a cardiac organoid using the device of the present invention. Specifically, it illustrates two probes (110-1', 110-2') being inserted into the cardiac organoid to measure bioimpedance.

[0176] The bioimpedance measured as described above can be transmitted to the signal processing unit via the measurement unit, and the signal processing unit can profile it in real time. An example of data actually acquired according to this method is shown in Fig. 9.

[0177] The data in Fig. 9 are profiling of impedance measured from two probes (110-1', 110-2'). Meanwhile, the data shown in Fig. 9 are profiling of impedance data measured from electrodes located along the extension direction of the probes (110-1', 110-2') from the ends of the probes (110-1', 110-2') from the bottom to the top (i.e., located along the direction toward the driving unit (120) described with reference to Figs. 1 and 2).

[0178] Referring to the data in Fig. 9, it can be confirmed that the impedances are profiled according to the depth depending on the position where the probes (110-1', 110-2') are inserted, and sections where the impedance changes rapidly at specific points can be identified. These sections of rapid change can be used as boundaries to compare the electrophysiological and drug responses by section. Meanwhile, based on these sections of rapid change, it is possible to verify in real time how deeply the probes (110-1', 110-2') are inserted within the 3D biomodel, or whether they are inserted to an appropriate degree.

[0179] FIG. 10 is a schematic diagram schematically showing an analysis of a retinal organoid using a device according to another embodiment of the present invention.

[0180] FIG. 11 is an image showing an example of in vivo spike profiling of a retinal organoid obtained using a device according to another embodiment of the present invention.

[0181] Hereinafter, in order to explain in detail the embodiments of FIGS. 10 and 11, the probe is illustrated with a separate drawing reference numeral (110-1'').

[0182] In the case of retinal organoids, there may be a specific layer within which biological spikes are detected. For example, referring to FIG. 9, the retinal ganglion cell (RGC) layer (RGCL) or the photoreceptor layer (PRL) may be examples thereof. Therefore, using the device of an embodiment of the present invention, a probe capable of measuring electrical signals according to depth within a 3D biomodel can be used to identify the coordinates of the target layer that emits biological spikes, and the maturity and responsiveness of the target layer can be determined, thereby allowing for analysis of intercellular conduction and networks based on this.

[0183] In Fig. 10, the method of analyzing a retinal organoid (RO) using the device of the present invention is roughly simulated. Specifically, the method of inserting a probe (110-1'') into the retinal organoid (RO) and measuring biopotential is simulated.

[0184] The biopotential measured as described above can be transmitted to the signal processing unit via the measurement unit. The signal processing unit can process the transmitted biopotential-related signal to profile the biospike. An example of an actual profiled biospike according to this aspect is shown in Fig. 11.

[0185] Among the graphs in Fig. 11, the graphs indicated as Ch are the profiles of the biopotential measured through the electrode(s) of the probe (110-1'') located near the ganglion cell layer (RGCL) inside the organoid.

[0186] Referring to the graphs of Fig. 11, it can be confirmed that among the electrodes of the probe (110-1''), the electrodes whose biological spikes are profiled are different from the specific electrodes (CH-03 of Fig. 11). By comparing the insertion location and depth of the retinal organoid (RO) of the probe (110-1''), the location and coordinates of the target layer can be identified based on the profiled information, and the maturity, responsiveness, etc. of the target layer can also be analyzed.

[0187] Figure 12 is a schematic diagram schematically showing an analysis of the bio-ion concentration of an organoid using a device according to another embodiment of the present invention.

[0188] The embodiment of the device shown in Fig. 12 is somewhat different from the embodiments of the device shown in the previous drawings. The device of the embodiment of Fig. 12 can be applied with the probe (110') of the embodiment described with reference to Fig. 5. Meanwhile, the probe part may further include a reference electrode (Ref) in addition to the probe (110'), the fixing part (400) may further include an electrolyte (EL), and the three-dimensional biomodel (50) may be positioned while being impregnated in the electrolyte (EL).

[0189] As described above with reference to FIG. 5, the probe (110') further includes one or more ion-selective membranes (112) covering the surfaces of at least some of the electrodes (111) among the plurality of electrodes (111), and as an example, as shown in FIG. 5, the plurality of electrodes (111) include the first group (111A) and the second group (111B), wherein the surface of one of the first group (111A) and the second group (111B) may be covered by the one or more ion-selective membranes (112). The following examples may be referred to together with FIG. 5.

[0190] By using a device according to the above embodiment, even if all potential changes at the boundary surface of the three-dimensional biomodel (50) are not known, a specific ion concentration inside an arbitrary three-dimensional biomodel (50) can be estimated or measured with high accuracy.

[0191] FIG. 12 illustrates a method of analyzing a three-dimensional biomodel (50) using the device of the present invention. Referring to FIG. 12, the probe (110') is inserted into the three-dimensional biomodel (50). The surfaces of the electrodes (111) constituting the second group (111B) among the electrodes (111) of the probe (110') are exposed, and the surfaces of the electrodes (111) constituting the first group (111A) are covered by the ion-selective membrane (112). Meanwhile, the first group (111A) and the second group (111B) are symmetrical with respect to any one of the planes including the direction (DRE).

[0192] The potential can be measured from the electrodes constituting the first group (111A) and the second group (111B), respectively. The potential can be transmitted to the signal processing unit via the measurement unit, and the signal processing unit can analyze the bio-ion concentration based on the potential. Specifically, the analysis can be performed using the following method.

[0193] The potential measured at the electrode (111) constituting the first group (111A) can satisfy Equation 1 below.

[0194] [Formula 1]

[0195] E ISE = E LJP + E LFP + E ref + E mem

[0196] In the above equation 1, E ISE is the potential measured at the electrodes constituting the first group, E LJP Liquid Junction potential, E, in a 3D biomodelLFP Liquid Field potential, E, in a 3D biomodel ref is the potential of the reference electrode (ref), E mem refers to the potential difference between the ion-selective membrane (112) and the internal solution of the three-dimensional biomodel (50).

[0197] The potential measured at the electrode (111) constituting the second group (111B) can satisfy Equation 2 below.

[0198] [Formula 2]

[0199] E Bare = E LJP + E LFP + E ref

[0200] In the above equation 2, E Bare is the potential measured at the electrodes constituting the second group, E LJP , E LFP and E ref is defined as in Equation 1.

[0201] As the electrodes (111) of the first group (111A) and the second group (111B) of the probe (110') are symmetrical to each other, the difference between the potential measured at the electrodes constituting the first group and the potential measured at the electrodes constituting the second group can be calculated. Therefore, by calculating using Equation 3 as described above, E can be easily obtained by simply obtaining the difference between the potential value measured at the electrodes constituting the first group and the potential value measured at the electrodes constituting the second group. mem can be obtained.

[0202] [Formula 3]

[0203] △E = E ISE - E Bare = E mem

[0204] Using the Nernst Equation, the above E mem The concentration of a specific ion (an ion selective to the ion-selective membrane) can be obtained from .

[0205] In this way, by using a device according to the above embodiment, even if all potential changes at the boundary surface of the three-dimensional biomodel (50) are not known, a specific ion concentration inside an arbitrary three-dimensional biomodel (50) can be estimated or measured with high accuracy.

[0206] In one embodiment, the control unit (300) may include an image processing unit (320) that processes information transmitted from the optical unit (200).

[0207] As in the above-described embodiment, the observation unit (210) of the optical unit (200) can obtain optical information of the three-dimensional biomodel (50) in real time.

[0208] The optical information of the three-dimensional bio-model (50) acquired from the observation unit (210) may be transmitted to and processed by the image processing unit (320). The image processing unit (320) may extract the coordinates of the projection position of the probe (110) from the optical information transmitted from the observation unit (210) and visualize them. Meanwhile, the extracted coordinates may be matched to mechanical coordinates built into the device (10). Based on this, the image processing unit (320) may quantitatively provide the real-time position of the probe (110) or visually provide the real-time position to the user.

[0209] Based on the above information, the user can verify in real time the process of inserting the probe (110) into the 3D biomodel (50). In addition, the above information is transmitted to the control unit (330) described later, and the control unit (330) can control the probe (110) to be accurately inserted into the 3D biomodel (50) based on the real-time information received. Alternatively, the probe (110) can be assisted to be accurately inserted into the 3D biomodel (50) by the user's operation.

[0210] In one embodiment, the control unit (300) may include a control unit (330) that controls the operation of the probe unit (100) and the optical unit (200) based on information processed from at least one of the signal processing unit (310) and the image processing unit (320).

[0211] In one embodiment, the control unit (330) can control the operation of the probe unit (100) and the optical unit (200) based on information processed from at least one of the signal processing unit (310) and the image processing unit (320).

[0212] In one embodiment, the control unit (330) can control the operation of the probe unit (100) and the optical unit (200) based on the information processed by the signal processing unit (310). In an exemplary embodiment, the operation of the drive unit (120), the operation of the electrical stimulation unit (140), or the operation of the optical stimulation unit (220) can be controlled based on the information processed by the signal processing unit (310).

[0213] In an exemplary embodiment, based on the local biopotential profile and bioimpedance profile transmitted from the signal processing unit (310), it can be determined during the analysis process whether the probe (110) is correctly positioned within the three-dimensional biomodel (50), how much it is inserted, or whether it is inserted to an appropriate degree. If the probe (110) is incorrectly positioned within the three-dimensional biomodel (50), is not inserted to a targeted depth, or is not inserted to an appropriate degree, the control unit (330) can adjust the position, etc., of the probe (110) by controlling the drive unit (120) based on the above information.

[0214] In an exemplary embodiment, it is possible to determine whether the state of the 3D biomodel (50) to be analyzed is normal or abnormal based on the local biopotential profile, bioimpedance profile, and biospike profile transmitted from the signal processing unit (310). The control unit (330) can compare the profiles with preset conditions to determine whether to apply an electrical stimulus and / or an optical stimulus to the 3D biomodel (50). If it is determined that the electrical stimulus and / or the optical stimulus should be applied, the control unit (330) can control the electrical stimulus unit (140) and / or the optical stimulus unit (220) to apply the electrical stimulus and / or the optical stimulus to the 3D biomodel (50).

[0215] Meanwhile, in an exemplary embodiment, the above judgment may be continuously performed even while an electrical stimulus or an optical stimulus is being applied. If it is determined that the state of the three-dimensional biomodel (50) has become normal due to the application of the electrical stimulus and / or optical stimulus as described above, the control unit (330) may control the electrical stimulus unit (140) and / or the optical stimulus unit (220) to terminate the application of the electrical stimulus and / or optical stimulus to the three-dimensional biomodel (50).

[0216] In an exemplary embodiment, based on the information transmitted from the image processing unit (320), it can be determined during the analysis process whether the probe (110) is correctly positioned within the three-dimensional biomodel (50), how much it is inserted, or whether it is inserted to an appropriate degree. If the probe (110) is incorrectly positioned within the three-dimensional biomodel (50), is not inserted to a targeted depth, or is not inserted to an appropriate degree, the control unit (330) can adjust the position, etc., of the probe (110) by controlling the drive unit (120) based on the above information.

[0217] In one embodiment, the information transmitted to the control unit (330) as described above may also be provided to the user via a display unit (not shown) or the like. The user may directly perform the aforementioned control based on the provided information, and in this case, the control unit (330) may perform the same control according to the user's operation.

[0218] In one embodiment, each component of the control unit (300) or the configuration of the storage unit (not shown) to be described later may be implemented by hardware, software, or a combination of hardware and software.

[0219] In an exemplary embodiment, the hardware may be implemented as an Application Specific Integrated Circuit (ASIC), Digital Signal Processing (DSP), Programmable Logic Device (PLD), Field Programmable Gate Array (FPGA), processor, controller, microprocessor, other electronic unit, or a combination thereof designed to perform the functions described above.

[0220] Meanwhile, in an exemplary embodiment, the software may be implemented as a module that performs the functions described above. The software may be stored in a memory unit and executed or operated by a processor. The memory unit or processor may employ various known means without limitation.

[0221] Examples of the above memory unit include a combination of non-volatile memory such as a flash memory disk (Solid State Disc, SSD), a hard disk drive, flash memory, EEPROM (Electrically Erasable Programmable Read-Only Memory), SRAM (Static RAM), FRAM (Ferro-electric RAM), PRAM (Phase-change RAM), MRAM (Magnetic RAM), and / or volatile memory such as DRAM (Dynamic Random Access Memory), SDRAM (Synchronous Dynamic Random Access Memory), and DDR-SDRAM (Double Date Rate-SDRAM).

[0222] In one embodiment, the device (10) may further include a display unit (not shown). Information or data produced within the device (10) may be displayed visually or audibly through the display unit (not shown).

[0223] In one embodiment, the device (10) may further include a storage unit (not shown). Information or data produced within the device (10) may be stored in the storage unit (not shown).

[0224] As described above, the device according to one embodiment of the present invention can provide a platform capable of analyzing 3D biomodels, as well as comprehensively interpreting and providing feedback on data that can be measured differently for each 3D biomodel. Furthermore, the device can provide a platform capable of comprehensively controlling the entire analysis process, from probe insertion into the 3D biomodel being analyzed and quality control of the 3D biomodel, to post-drug treatment analysis.

[0225] Figure 13 is a step diagram showing an example of a method for analyzing a three-dimensional biomodel according to one embodiment of the present invention.

[0226] The method for analyzing a three-dimensional biomodel according to one aspect of the present invention can be performed using the device for analyzing a three-dimensional biomodel according to one aspect of the present invention described above with reference to FIGS. 1 to 12. Hereinafter, redundant descriptions of each component of the device (10) described with reference to FIGS. 1 to 12 will be omitted.

[0227] In one embodiment, a method for analyzing a three-dimensional biomodel may include a step of inserting the probe into the three-dimensional biomodel (S10); a step of verifying a process of inserting the probe into the three-dimensional biomodel (S20); a step of obtaining an electrical signal of the three-dimensional biomodel (S30); a step of applying an electrical stimulus or an optical stimulus to the three-dimensional biomodel (S40); and a step of determining whether an electrical signal measured from the three-dimensional biomodel satisfies a preset condition (S50).

[0228] In one embodiment, step S10 may insert the probe into the three-dimensional biomodel. As described above, the probe may be inserted into the three-dimensional biomodel along the extension direction.

[0229] Meanwhile, in one embodiment, step S10 may be automatically performed by the driving unit (120). Step S10 may be performed in real time by providing feedback based on information transmitted from the probe unit (100) and the optical unit (200).

[0230] In one embodiment, the step S20 may verify the process of inserting the probe into the three-dimensional biomodel.

[0231] As described above, based on the local biopotential profile and bioimpedance profile transmitted from the signal processing unit and the information transmitted from the image processing unit, it is possible to determine in real time whether the probe is correctly positioned within the 3D biomodel, how much it is inserted, or whether it is inserted to an appropriate degree during the analysis process. Meanwhile, while determining the above in real time, the position of the probe can be adjusted if necessary. The adjustment of the position of the probe can be performed automatically by the control unit described above, or can be performed by a user's operation through the control unit.

[0232] In one embodiment, in step S30, an electrical signal of the three-dimensional biomodel can be obtained.

[0233] As described above, the step S30 can be performed by obtaining an electrical signal of the three-dimensional biomodel from the measurement unit of the probe, and the obtained electrical signal can be processed in the signal processing unit.

[0234] Meanwhile, the step S30 may be performed between the steps S20 and S40, but may also be performed in parallel with at least some of the steps S10, S20, S40, and S50. For example, the step S30 may be performed in parallel with all of the steps S10, S20, S40, and S50. In such an embodiment, the electrical signal within the 3D biomodel may be acquired simultaneously with inserting the probe into the 3D biomodel, and the electrical signal within the 3D biomodel may be continuously acquired even during the start or end process of applying electrical stimulation and / or optical stimulation to the 3D biomodel, and further, the electrical signal within the 3D biomodel may be continuously acquired even after the end of applying the electrical stimulation and / or optical stimulation.

[0235] Figure 14 is a graph showing an example of stabilizing a cardiac organoid by applying electrical stimulation.

[0236] Figure 15 is a graph showing the biopotential profiles 3 minutes (A), 4 minutes (B), 5 minutes (C), and 6 minutes (D) after inserting the probe into the cardiac organoid.

[0237] In one embodiment, in step S40, an electrical stimulus or an optical stimulus may be applied to the three-dimensional biomodel. The electrical stimulus may be performed by an electrical stimulus unit of the probe portion via the probe, and the optical stimulus may be performed by an optical stimulus unit of the optical portion.

[0238] In one embodiment, step S40 may apply electrical stimulation to the three-dimensional biomodel. In an exemplary embodiment, the three-dimensional biomodel to which the electrical stimulation is applied may be a cardiac organoid.

[0239] In an exemplary embodiment, step S40 may be performed to establish quality control of the cardiac organoid to be analyzed.

[0240] As previously described, cultured cardiac organoids may exhibit abnormal behavior or conditions, such as irregular heartbeats or failure to reach a normal heart rate, due to stress caused by changes in the initial environment during analysis.

[0241] Fig. 14 shows the analysis of the beating rate over time, with the time point of probe insertion into the cardiac organoid set to 0. Meanwhile, Fig. 15 shows the biopotential profiles 3 minutes (A), 4 minutes (B), 5 minutes (C), and 6 minutes (D) after probe insertion into the cardiac organoid, respectively. The graph of Fig. 14 may be a graph created based on the profiles of Fig. 15. Meanwhile, the above measurement may be performed by the aforementioned step S30.

[0242] Referring to Figures 14 and 15, upon probe insertion into a cardiac organoid on day 32 of culture, the analyzed cardiac organoid initially exhibited an abnormally slow heart rate and irregular beating (~5 minutes) due to inactivation. This may indicate that the analyzed cardiac organoid was in an abnormal state. To perform accurate bioanalysis (e.g., drug testing) using cardiac organoids, the analyzed cardiac organoid must be in a normal state.

[0243] In order to stabilize and normalize the state of the cardiac organoid to be analyzed, 5 minutes after the probe was inserted into the cardiac organoid, electrical stimulation was applied to the cardiac organoid via the probe.

[0244] In one embodiment, the electrical stimulation may be a current pulse or voltage pulse in the form of a pulse train or a single pulse.

[0245] In one embodiment, the electrical stimulation may be a current pulse or voltage pulse in the form of a single pulse.

[0246] In one embodiment, the electrical stimulation may be a current pulse or voltage pulse in the form of a pulse train.

[0247] In one embodiment, both current or voltage pulses in the form of pulse trains and current or voltage pulses in the form of single pulses may be applied. For example, the current or voltage pulses in the form of pulse trains and current or voltage pulses in the form of single pulses may be applied sequentially, or at least some of them may be applied in parallel.

[0248] In one embodiment, the pulse width of the pulse may be 0.1 ms to 1 ms.

[0249] In one embodiment, the frequency of the pulse train may be from 0.1 Hz to 10 Hz.

[0250] In one embodiment, the current pulse may have a peak of 1 nA to 1 mA.

[0251] In one embodiment, the electrical stimulation may be a current pulse in the form of a pulse train. In this case, at least one of the following conditions may be satisfied.

[0252] a. Pulse width is 0.1 ms to 1 ms,

[0253] b. The frequency of the pulse train is 0.1 Hz to 10 Hz,

[0254] c. Peak is 1nA to 1mA.

[0255] In one embodiment, the current pulse in the form of a pulse train may satisfy all of the conditions a to c above.

[0256] In one embodiment, the application of the electrical stimulation may be performed for 1 second to 10 minutes.

[0257] When applying electrical stimulation as described above, stabilization of cardiac organoids by electrical stimulation may be more effective.

[0258] Referring again to Figure 14, 5 minutes after the probe was inserted into the cardiac organoid, an electrical stimulus in the form of a pulse train (1 μA, 10 pulse train, 500 ms) was applied to the cardiac organoid via the probe.

[0259] Referring to FIGS. 14 and 15, it was confirmed that the heart rate of the cardiac organoid was normalized and stabilized (the dispersion of the bpm value dropped to the level of 1 to 2 bpm) after the application of the electrical stimulation as described above.

[0260] In one embodiment, step S40 may apply a light stimulus to the three-dimensional biomodel. In an exemplary embodiment, the three-dimensional biomodel to which the light stimulus is applied may be a retinal organoid.

[0261] In an exemplary embodiment, step S40 may be performed to establish quality control of the retinal organoids to be analyzed.

[0262] Retinal organoids in a normal (healthy) state can respond to light stimulation. On the other hand, retinal organoids in an abnormal state may not respond to light stimulation or may respond to light stimulation to a low degree. To perform accurate bioanalysis (such as drug testing) using retinal organoids, the state of the retinal organoids to be analyzed must be normal. In step S40, after applying light stimulation to the retinal organoids, the response of the retinal organoids to the light stimulation can be observed to determine whether the retinal organoids to be analyzed are normal or abnormal. Meanwhile, the response of the retinal organoids to the light stimulation can be detected based on the biological spike profile of the organoids.

[0263] In one embodiment, the light stimulation may be applied by a light source, as in the aforementioned embodiment. In an exemplary embodiment, the light source may continuously irradiate light to the periphery. In an exemplary embodiment, when applying the light stimulation by the light source, the stimulation may be applied by exposing the retinal organoid to the light source for a certain period of time, followed by turning off the light source or storing the retinal organoid in a dark space.

[0264] In one embodiment, the optical stimulus may be pulsed light in the form of a pulse train or a single pulse.

[0265] In one embodiment, the light stimulus may be pulsed light in the form of a single pulse.

[0266] In one embodiment, the optical stimulus may be pulsed light in the form of a pulse train.

[0267] In one embodiment, both pulsed light in the form of a pulse train and pulsed light in the form of a single pulse may be applied. For example, the pulsed light in the form of a pulse train and pulsed light in the form of a single pulse may be applied sequentially, or at least some of them may be applied in parallel.

[0268] In one embodiment, the pulse width of the pulsed light may be from 0.01 ms to 1,000 ms.

[0269] In one embodiment, the intensity of the pulsed light is 0.01 mW / cm 2 1,000mW / cm 2 It could be.

[0270] In one embodiment, the wavelength of the light may be 300 nm to 800 nm.

[0271] In one embodiment, the pulsed light may satisfy at least one of the following conditions:

[0272] a. Pulse width of 0.01ms to 1,000ms,

[0273] b. Intensity is 0.01 mW / cm 2 1,000mW / cm 2 ,

[0274] c. Wavelength is 300 nm to 800 nm.

[0275] In one embodiment, the pulsed light may satisfy all of the conditions a through c above.

[0276] In one embodiment, the application of the optical stimulus may be performed for 1 second to 10 minutes.

[0277] When applying the above-described optical stimulation, the feedback of retinal organoids responding to the optical stimulation can be observed more easily and effectively.

[0278] Through the above step S40, the state of the cardiac organoids to be analyzed can be normalized and stabilized, and abnormal cardiac organoids can be identified. Meanwhile, through the above step S40, abnormal retinal organoids among the retinal organoids to be analyzed can be identified.

[0279] Referring again to FIG. 13, in one embodiment, in step S50, it can be determined whether the electrical signal measured from the three-dimensional biomodel satisfies a preset condition.

[0280] As in the above-described embodiment, step S30 may also be performed in parallel with step S40. At this time, if the electrical signal measured from the 3D biomodel satisfies a preset condition, this may indicate normalization or stabilization of the 3D biomodel, or may indicate that the status determination of the 3D biomodel has been completed.

[0281] In one embodiment, step S50 may determine whether the electrical signal measured from the three-dimensional biomodel satisfies a preset condition. In one embodiment, the three-dimensional biomodel may be a cardiac organoid.

[0282] In one embodiment, the condition used to determine whether to continue applying the electrical stimulation may be whether each variation of the amplitude, beating rate, FPD (Field potential duration), ERP (Effective Refractory Period), CS (conduction speed), etc. of the cardiac organoid's beating for 1 minute is within 25%. That is, if the variation of the amplitude of the cardiac organoid for 1 minute after the application of the electrical stimulation by the step S40 is within 25%, the variation of the beating rate is within 25%, and the variation of the FPD is within 25%, the application of the electrical stimulation may be terminated. A cardiac organoid that satisfies the above conditions may be regarded as a cardiac organoid in a normal state.

[0283] Meanwhile, if it is determined that the electrical signal satisfies the preset conditions, the application of the electrical stimulation can be terminated.

[0284] In one embodiment, step S50 may determine whether the electrical signal measured from the three-dimensional biomodel satisfies a preset condition. In one embodiment, the three-dimensional biomodel may be a retinal organoid.

[0285] In one embodiment, the condition used to determine whether the application of the optical stimulus continues may be whether the firing rate of the RGC spike of the retinal organoid is 2 Hz or higher, or whether the optical stimulus delay is 100 ms or lower. Here, in the case of the RGC spike, a signal having a signal-to-noise ratio (SNR) of 10 dB or higher may be determined as a threshold.

[0286] That is, if the firing rate of the RGC spike of the retinal organoid is 2 Hz or more and the light stimulus delay is 100 ms or less after the light stimulus is applied in step S40, the light stimulus application can be terminated. A retinal organoid that satisfies the above conditions can be regarded as a normal retinal organoid.

[0287] Meanwhile, if it is determined that the electrical signal satisfies the preset conditions, the application of the optical stimulus can be terminated.

[0288] Based on the above conditions, the status of cardiac organoids and retinal organoids can be determined more accurately.

[0289] In one embodiment, steps S10 through S50 optimize analysis conditions for a 3D biomodel, and establish quality control. By performing these steps, biotests, such as drug treatment tests, can be accurately and effectively performed on the 3D biomodel being analyzed.

[0290] In one embodiment, the three-dimensional biomodel (50) can be analyzed while maintaining its original shape.

[0291] In one embodiment, the three-dimensional biomodel (50) can be examined and analyzed non-destructively. In an exemplary embodiment, the three-dimensional biomodel (50) can be examined and analyzed with minimal invasion.

[0292] Therefore, analysis while maintaining the original shape can mean that analysis is performed without causing deformation of the shape in the remaining parts except for the part invaded by the probe.

[0293] Conventional analytical devices and methods, particularly those utilizing 2D electrodes, require the 3D biomodel being analyzed to be sectioned to secure a cross-section before conducting bioanalysis or quality control analysis. Other methods inevitably involve some degree of destruction of at least a portion of the 3D biomodel. In such cases, it can be difficult to ensure the accuracy of biotest results using the 3D biomodel, or the analysis can be challenging or the protocol can be cumbersome.

[0294] According to the method for analyzing a 3D biomodel according to an embodiment of the present invention, the 3D biomodel can be non-destructively analyzed while maintaining its original shape. Accordingly, the accuracy of biometric test results can be improved, and the analysis method can be made more efficient.

[0295] Although the present invention has been described through the embodiments of the present invention as described above, these are provided only to help a more general understanding of the present invention, and the present invention is not limited to the above examples, and those skilled in the art to which the present invention pertains can make various modifications and variations based on these descriptions.

[0296] Accordingly, the idea of ​​the present invention is that not only the following claims but also all modifications equivalent to or equivalent to the claims fall within the scope of the idea of ​​the present invention.

Claims

As an analysis device for 1.3D biomodels, A probe unit that probes the above 3D biomodel to obtain an electrical signal and applies an electrical stimulus to the above 3D biomodel; An optical unit for optically observing the three-dimensional biomodel and applying optical stimulation to the three-dimensional biomodel; and A control unit that controls the probe unit and the optical unit; The above probe portion includes a probe extending in one direction, The above probe is a device comprising a plurality of electrodes arranged along the above direction.

2. In paragraph 1, The above 3D biomodel is, A device comprising at least one selected from the group consisting of cardiac organoids, retinal organoids, brain organoids, tumor organoids, neural organoids, and organoids comprising cells subjected to optogenetic manipulation.

3. In paragraph 1, A device having a plurality of the above probes.

4. In paragraph 1, The above probe, A device further comprising one or more ion-selective membranes covering the surfaces of at least some of the electrodes among the plurality of electrodes.

5. In paragraph 4, The above plurality of electrodes, It is classified into a first group arranged along the direction on one side of the probe and a second group arranged along the direction on the other side of the probe, A device, wherein one of the first and second groups has a surface covered by one or more ion-selective membranes.

6. In paragraph 1, The above probe part, A device further comprising a driving unit for controlling the position of the probe.

7. In paragraph 1, The above probe part, A device further comprising a measuring unit that measures electrical signals obtained from at least some of the plurality of electrodes of the probe.

8. In paragraph 7, The above measurement unit is a device that measures electrical signals according to depth within the three-dimensional biomodel.

9. In paragraph 7, The above electrical signal is, A device comprising at least one of a biopotential and a bioimpedance of the three-dimensional biomodel.

10. In paragraph 1, The above probe part, A device further comprising an electrical stimulation unit that applies electrical stimulation to the three-dimensional biomodel via the probe.

11. In paragraph 10, The device wherein the electrical stimulation is a current pulse or voltage pulse in the form of a pulse train or a single pulse.

12. In paragraph 1, The above optical part, A device comprising an observation unit that acquires optical information of the three-dimensional biomodel in real time.

13. In paragraph 1, The above optical part, A device comprising a light stimulation unit that applies light stimulation to the above three-dimensional biomodel.

14. In paragraph 13, The above optical stimulus is a pulsed light in the form of a pulse train or a single pulse, the device.

15. In paragraph 1, The above control unit, A device comprising a signal processing unit that processes information transmitted from the probe unit and an image processing unit that processes information transmitted from the optical unit.

16. In paragraph 15, The above control unit, A device comprising a control unit that controls the operation of the probe unit and the optical unit based on information processed from at least one of the signal processing unit and the image processing unit.

17. A method for analyzing a three-dimensional biomodel using the device according to Article 1, A step of inserting the probe into the three-dimensional biomodel; A step of verifying the process of inserting the probe into the three-dimensional biomodel; A step of acquiring an electrical signal of the above three-dimensional biomodel; A step of applying electrical stimulation or optical stimulation to the above three-dimensional biomodel; and A method comprising a step of determining whether an electrical signal measured from the three-dimensional biomodel satisfies a preset condition.

18. In paragraph 17, The above electrical stimulation is a current pulse in the form of a pulse train, The above current pulse satisfies at least one of the following conditions: a. Pulse width is 0.1 ms to 1 ms, b. The frequency of the pulse train is 0.1 Hz to 10 Hz, c. Peak is 1nA to 1mA.

19. In paragraph 17, The above light stimulus is pulsed light, The above pulsed light satisfies at least one of the following conditions: a. Pulse width of 0.01ms to 1,000ms, b. Intensity is 0.01 mW / cm 2 1,000mW / cm 2 , c. Wavelength is 300 nm to 800 nm.

20. In paragraph 17, A method in which the above three-dimensional biomodel is analyzed while maintaining its original shape.

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