Semiconductor mesh for cell barrier analysis

Abstract

Semiconductor mesh for cell barrier analysis. This patent application relates to a semiconductor cell culture apparatus. The apparatus includes a semiconductor mesh having interconnected islands and vias, a cap defining a space for a cell culture, mesh and cap electrodes, and electrical connections. The electrical connections allow electrical measurements between the mesh and cap electrodes. The mesh electrodes must satisfy the geometric condition that at least one conductive point must be located within a specific distance from the via. The apparatus allows for defect detection in biological barriers, wherein the electrical measurements provide an output indicating any defects. The apparatus may be part of a system including a processing unit suitable for performing defect detection methods.

Description

Technical Field

[0001] This invention relates to the fields of biomedical engineering and cell culture devices, and more specifically, to semiconductor cell culture devices and methods for detecting defects in biological barriers. Background Technology

[0002] Cell culture and barrier integrity analysis are key areas of research in biomedical science. This field is particularly relevant to the study of biological barriers such as the blood-brain barrier (BBB), intestinal barrier, lung barrier, and liver barrier. These barriers play a vital role in protecting the body from harmful substances, and their integrity is essential for maintaining health.

[0003] One of the main methods for studying these barriers and their integrity is through the use of cell culture devices. These devices typically contain a mesh structure that forms the basis for cell layer growth. The mesh structure usually includes through-holes, which are spaces between mesh islands in which cells can grow and form barriers.

[0004] A major challenge in this field is the accurate measurement of barrier integrity, such as the detection of microscopic gaps in the barrier. Traditional methods, such as transepithelial-transendothelial resistance (TEER) measurements, have been used to assess barrier integrity. These methods involve placing electrodes at the top and bottom of the device chamber and measuring the resistance between them. However, these methods have limitations. For example, they may not provide a detailed, localized assessment of barrier integrity at the level of individual pores. Furthermore, traditional methods may not be sensitive enough to detect small gaps that could significantly impact barrier function.

[0005] Despite advancements in cell culture apparatus and methods for barrier integrity analysis, significant challenges remain in this field. Further progress is needed to address these challenges and improve the accuracy and sensitivity of barrier integrity measurements. Summary of the Invention

[0006] The objective of this invention is to provide an apparatus and method capable of performing robust barrier integrity analysis (particularly for the blood-brain barrier) by enabling the detection of gaps at the microscopic level. This objective is achieved through the semiconductor cell culture apparatus, system, and method according to the invention.

[0007] In a first aspect, the present invention relates to a semiconductor cell culture apparatus, comprising: a semiconductor grid having interconnected islands and through-holes; a cap facing the grid and defining a cell culture space; at least one grid electrode attached to the grid; at least one cap electrode attached to the cap; and first and second electrical connections (111) to the grid and the cap electrode, respectively, said electrical connections allowing electrical measurements between the electrodes. The grid electrode must satisfy the following geometric condition: at least one conductive point must be located within 100 µm of the through-hole. 100 µm is the radius of the largest endothelial / epithelial cell in the culture.

[0008] Preferably, it relates to a semiconductor cell culture apparatus, comprising:

[0009] a. A semiconductor mesh (112) having islands (116) interconnected by bridges (118) and defining vias (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface. b. A cap, which faces and is spaced apart from the top surface of the semiconductor grid (112), thereby defining a space for cell culture, wherein the cap has a top surface and a bottom surface. c. At least one grid electrode attached to the top or bottom surface of the grid. d. At least one cover electrode attached to the bottom surface of the cover. e. A first electrical connection (110) to one or more of the at least one grid electrode, and f. A second electrical connection (111) to one or more of the at least one cap electrode. The first and second electrical connections allow for electrical measurements between the one or more of the at least one grid electrode and the cap electrode, and The at least one or more of the grid electrodes satisfy a geometric condition such that at least one conductive point of each of the at least one or more of the grid electrodes must be located within a distance (d) of 100 µm from the via (114).

[0010] In a second aspect, the present invention relates to a method for detecting the presence of defects in a biological barrier in a semiconductor cell culture apparatus according to any embodiment of the first aspect. The method includes sequentially initiating electrical measurements between each grid electrode and cap electrode of the apparatus, generating an electrical output, and determining from the output that if any grid electrode measures an output outside a predetermined range, it indicates the presence of a defect in the cell culture above a via within the distance between these electrodes.

[0011] Preferably, it can relate to a method for detecting the presence of defects in a biological barrier in a semiconductor cell culture apparatus according to any one of the preceding claims, the method comprising:

[0012] a. An electrical measurement is sequentially initiated between each of one or more of the at least one grid electrode and the cap electrode of the semiconductor cell culture device, thereby generating an electrical output. b. Based on the electrical output obtained from electrical measurements, if any one or more of the at least one grid electrode measures an output outside a predetermined range, it indicates a defect in the cell culture above the through-hole (114) within the distance (d) of these electrodes.

[0013] In a third aspect, the present invention relates to a semiconductor cell culture system, including apparatus according to any embodiment of the first aspect, and further including a processing unit adapted to perform the method of the second aspect.

[0014] In a fourth aspect, the present invention relates to a computer program comprising instructions for causing a semiconductor cell culture system of the third aspect to perform the steps of the method of the second aspect.

[0015] In a fifth aspect, the present invention relates to a computer-readable medium having a computer program of the fourth aspect stored thereon.

[0016] An advantage of this invention is that it enables a semiconductor cell culture device that allows for precise and accurate measurement of barrier integrity at the microscopic level. Another advantage is that the device can identify pore defects, thereby improving the accuracy of notch detection.

[0017] Another advantage of embodiments of the invention is that the device can detect defects in the barrier, particularly at the pore level, such as voids, irregularities, cracks, inhomogeneities, or inconsistent tightness. This feature is particularly useful in drug delivery and barrier integrity analysis because it allows for clear observation and electrical measurement of barrier integrity.

[0018] Another advantage of this invention is that the device can be integrated into a microfluidic chip. This integration simplifies and improves efficiency, saving researchers and scientists time and resources.

[0019] Specific and preferred aspects of the invention are set forth in the appended independent and dependent claims. Features from the dependent claims may be suitably combined with features of the independent claims and other dependent claims, not merely as expressly set forth in the claims.

[0020] The above and other features, characteristics, and advantages of the present invention will become apparent from the following detailed description taken in conjunction with the accompanying drawings, which illustrate the principles of the invention by way of example. This description is given for illustrative purposes only and does not limit the scope of the invention. The accompanying drawings are referenced below. Attached Figure Description

[0021] Figure 1 This is a top view of a horizontal cross-section of a semiconductor cell culture apparatus according to the prior art, showing a semiconductor mesh with islands interconnected by bridges and defining through-holes.

[0022] Figure 2 This is a schematic diagram of a vertical cross-section of a semiconductor cell culture apparatus according to the prior art, showing a lid facing the top surface of a semiconductor grid and defining a cell culture space, and electrodes at the lid and bottom to allow TEER.

[0023] Figure 3 This is a detailed top view schematically representing a portion of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing the geometry of the grid electrodes relative to the vias.

[0024] Figure 4 This is a schematic diagram of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing an electrical connection between two grid electrodes via a switch.

[0025] Figure 5 This is a schematic diagram of a vertical cross-sectional view of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing a cover facing the top surface of a semiconductor grid and defining a space for cell cultures, and electrodes on the bottom surface of the cover and the top of the grid to allow for electrical measurements.

[0026] Figure 6 It shows that it is attributed to Figure 5 The leakage in the embodiment is caused by defects in cell culture above the through-hole.

[0027] Figure 7 It shows that in relation to Figure 6 and 7 The path of charge when performing electrical measurements in a device similar to the one shown.

[0028] Figure 8 This is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing a cover facing the top surface of a semiconductor grid and defining a space for cell cultures, with a single large electrode on the bottom surface of the cover and multiple electrodes on the top of the grid to allow for electrical measurements.

[0029] Figure 9 and10 This is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing a cover facing the top surface of a semiconductor grid and defining a cell culture space, and having a single large electrode pierced by a hole on the bottom surface of the cover and multiple electrodes on the top of the grid to allow electrical measurements and cell observation (e.g., spectroscopy or microscopy).

[0030] Figure 11 This is a schematic diagram of a semiconductor cell culture apparatus, showing electrodes on the bottom surface of a grid, first and second electrical connections to the grid and cap electrodes respectively, an electrical measurement system connected to the first and second electrical connections, and a processing unit for performing a method according to an embodiment of the present invention.

[0031] Figure 12 This is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing a cover facing the top surface of a semiconductor grid and defining a space for cell cultures, and having a plurality of electrodes on the bottom surface of the cover and a plurality of electrodes on the bottom surface of the grid, the electrodes being at different distances from the through-holes.

[0032] Figure 13 and 14 This is a schematic top view of an electrode for attachment to the bottom surface of a cover according to an embodiment of the present invention.

[0033] Figure 15 This is a schematic diagram of a detailed top view of a grid electrode used in an embodiment of the present invention.

[0034] Figure 16 This is a schematic diagram of a detailed top view of the grid electrode (left) and the cap electrode (right) used in an embodiment of the present invention.

[0035] Figure 17 This is a flowchart illustrating a method for detecting the presence of defects in a biological barrier in a semiconductor cell culture apparatus according to an embodiment of the present invention.

[0036] In different accompanying drawings, the same reference numerals refer to the same or similar elements. Detailed Implementation

[0037] The invention will be described with reference to specific embodiments and certain accompanying drawings, but is not limited thereto; rather, it is limited only by the claims. The described drawings are merely illustrative and not restrictive. In the drawings, some elements may be enlarged and not drawn to scale for illustrative purposes. Dimensions and relative dimensions do not correspond to an actual reduction in scale for the practice of the invention.

[0038] The terms first, second, third, etc., used in the specification and claims are used to distinguish between similar elements and are not necessarily used to describe a temporal, spatial, hierarchical, or any other order. It is to be understood that the terms thus used are interchangeable where appropriate, and the embodiments of the invention described herein can operate in an order different from that described or illustrated herein.

[0039] Furthermore, terms such as top and above, etc., in the specification and claims are used for descriptive purposes and not necessarily to describe relative positions. It is to be understood that such terms are interchangeable where appropriate, and embodiments of the invention described herein can operate in orientations different from those described or illustrated herein.

[0040] It should be noted that the term "comprising" as used in the claims should not be construed as limiting itself to the means listed thereafter; it does not exclude other elements or steps. Therefore, the term should be interpreted as specifying the presence of the stated features, integers, steps, or components as mentioned, but does not exclude the presence or addition of one or more other features, integers, steps, or components, or groups thereof. Thus, the term "comprising" covers both cases where only the stated features are present and cases where these features are present along with one or more other features. The term "comprising" according to the invention therefore also includes an embodiment where no other components are present. Therefore, the scope of the statement "device comprising means A and B" should not be construed as limiting it to a device consisting only of components A and B. This means that, for the purposes of this invention, the relevant components in the device are only A and B.

[0041] Similarly, it should be noted that the term "coupling" should not be interpreted as limited to direct connection. The terms "coupling" and "connection" can be used together with their derivatives. It should be understood that these terms are not intended to be synonyms. Therefore, the scope of the phrase "device A coupled to device B" should not be limited to devices or systems where the output of device A is directly connected to the input of device B. This implies the existence of a path between the output of device A and the input of B, which can include other devices or apparatuses. "Coupling" can mean two or more elements in direct physical or electrical contact, or it can mean two or more elements that are not in direct contact but still cooperate or interact with each other.

[0042] Throughout this specification, the reference to "an embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of the invention. Therefore, the phrase "in one embodiment" or "in an embodiment" appearing in various places throughout this specification does not necessarily refer to the same embodiment, but may refer to the same embodiment. Furthermore, in one or more embodiments, as will be apparent to those skilled in the art from this disclosure, particular features, structures, or characteristics may be combined in any suitable manner.

[0043] Similarly, it should be understood that in the description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, drawing, or description for the purpose of simplification and to aid in understanding one or more of the various inventive aspects. However, this approach of the disclosure should not be construed as reflecting an intention to claim more features than are expressly recited in each claim. Rather, as reflected in the appended claims, inventive aspects lie in fewer features than all the features of a single foregoing disclosed embodiment. Therefore, the appended claims are thus explicitly incorporated into this detailed description, wherein each claim itself represents a separate embodiment of the invention.

[0044] Furthermore, while some embodiments described herein include features that are included in other embodiments but not others, it will be understood by those skilled in the art that combinations of features from different embodiments are intended to fall within the scope of the invention and form different embodiments. For example, any embodiment of the claimed embodiments in the appended claims may be used in any combination.

[0045] Furthermore, some embodiments described herein are described as methods or combinations of method elements that can be implemented by a processor of a computer system or by other means of implementing the functionality. Thus, a processor having the necessary instructions for performing the elements of such methods or methods forms means for performing the elements of methods or methods. Furthermore, the elements described herein in the apparatus embodiments are examples of means for implementing the functionality performed by elements that achieve the objectives of the present invention.

[0046] Numerous specific details are set forth in the description provided herein. However, it should be understood that embodiments of the invention can be practiced without these specific details. In other instances, well-known methods, structures, and techniques have not been shown in detail so as not to obscure the understanding of this specification.

[0047] The following items are provided separately to aid in understanding the invention.

[0048] The invention will now be described in detail through several embodiments thereof. It will be apparent that other embodiments of the invention can be configured based on the knowledge of those skilled in the art without departing from the technical teachings of the invention, which is limited only by the terms of the appended claims.

[0049] We now refer to Figure 1 . Figure 1 This is a top view of a horizontal cross-section of a semiconductor cell culture apparatus according to the prior art, showing a semiconductor grid (112) having islands (116) interconnected by bridges (118) and defining vias (114). Grid electrodes (120) are depicted at the center of the islands (116), away from the edges of any vias.

[0050] We now refer to Figure 2 . Figure 2 This is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture apparatus according to the prior art. It shows a cover (200) facing the top surface of a semiconductor grid (112) and defining a space (202) for cell culture (204) according to the prior art, and has a cover electrode (220) on the cover (200) and a bottom electrode (230) on the bottom of the apparatus to allow TEER. The cell culture (204) is depicted on the grid (112).

[0051] The device of the present invention allows for the identification of defects in biological barriers (e.g., cell cultures) at the pore level, i.e., the areas where the defects are most severe.

[0052] Figure 5 This is a schematic diagram of a vertical cross-section of a semiconductor cell culture apparatus according to the first aspect. As used herein, unless otherwise stated, the term "semiconductor cell culture apparatus" refers to an apparatus for the growth and maintenance of biological cells. The apparatus includes semiconductor materials. Figure 5 A cover (200) is shown facing and separated from the top surface of the semiconductor grid (112), thereby defining a space (202) for cell culture (204), wherein the cover has a top surface and a bottom surface. In other words, the cover is a component of the semiconductor cell culture apparatus that faces and is separated from the top surface of the semiconductor grid. The cover (200) also has a top surface and a bottom surface, and when located on the semiconductor grid, it defines a space (202) for cell culture (204).

[0053] According to an embodiment of the invention, the device further comprises at least one cap electrode (220) attached to the bottom surface of the cap (200) and at least one grid electrode (120) attached to the top surface of the grid (112) for allowing electrical measurements. The cap electrode is an electrode attached to the bottom surface of the cap (200). These electrodes are used for performing electrical measurements in a semiconductor cell culture apparatus. The grid electrode is also used for performing electrical measurements in a semiconductor cell culture apparatus. The grid electrode may also be attached to the bottom of the grid (see...). Figure 11 and 12 One or more of the at least one grid electrode (120) satisfy a geometric condition such that at least one conductive point (121) of each of the at least one or more of the at least one grid electrode (120) must be located within a distance (d) of 100 μm from the via (114).

[0054] A semiconductor mesh (112) has islands (116) interconnected by bridges (118) (not shown) and defines vias (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface. In other words, a semiconductor mesh refers to a network or fabric-like structure made of semiconductor material. The mesh includes interconnected islands and bridges and defines vias between the islands. The mesh has a top surface and a bottom surface. Island, bridge, and via are all components of the mesh structure. A first electrical connection (110) exists to one or more of the at least one mesh electrode (120), and a second electrical connection exists to one or more of the at least one cap electrode (220), but in Figure 5 Not shown in the diagram. The first and second electrical connections allow electrical measurements between one or more of the at least one grid electrode (120) and the cap electrode (220). The first electrical connection (110) is a connection to one or more grid electrodes. These connections allow the transmission of electrical signals between the grid electrodes and other components of the semiconductor cell culture apparatus. The second connection is a connection to one or more cap electrodes. These connections allow the transmission of electrical signals between the cap electrodes and other components of the semiconductor cell culture apparatus.

[0055] Figure 7 Electrical measurements are described herein. Electrical measurements are the process of determining the electrical properties of a cell culture (204). These measurements are typically performed sequentially between each of one or more of the at least one grid electrode and the cap electrode (220), facilitated by first and second electrical connections. Such electrical measurements allow for the detection of defects in the cell culture at the level of the through-hole (114). These defects are... Figure 6 The diagram is shown in which it is assumed that the chemicals confined in the space (202) have already passed through the defects in the cell culture (204) at the level of the pores (114).

[0056] Figure 3 This is a detailed top view, schematically representing a portion of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing the geometry of the grid electrodes (120) relative to the via (114). Unless otherwise stated, the term "geometric condition" as used herein refers to a specific spatial requirement that the grid electrodes must meet. This condition specifies that at least one conductive point (121) of one or more of the grid electrodes (120) must be located within a certain distance (d) from the via (114).

[0057] In various embodiments, the distance can be 10 μm from the via, preferably 5 μm. This embodiment allows for more accurate measurements as the electrode is closer to the via. Most preferably, the distance can be 1 µm, or even 0 µm (e.g., one or more of at least one conductive point from the first electrode and one or more of at least one conductive point from the second electrode must be in direct contact with or adjacent to the single via, such that there is essentially no distance between them and the via). In embodiments, the geometry may require each grid electrode (120) to have a conductive boundary with at least two distinct conductive points located within this distance and equidistant from the via. This is in Figure 3 As shown in the figure. This embodiment ensures the consistency of measurements.

[0058] In an embodiment, each grid electrode (120) may have a conductive boundary on its top surface, the conductive boundary having a line (122) at least 20 μm long and located entirely within the distance from the via (114). This also... Figure 3 As shown in the diagram, this line is enclosed by a dashed rectangle. This embodiment allows for a larger electrode area adjacent to the via, improving measurement accuracy.

[0059] In one embodiment, the line can be arranged parallel to the periphery of the through-hole, the periphery being coplanar with the surface of the grid electrode (120) attached to the grid. This also... Figure 3 The scenario is illustrated. This embodiment ensures that the electrodes are in the optimal position for accurate measurement.

[0060] In an embodiment, the geometry may be further specified: at least 50%, preferably at least 75%, more preferably at least 90%, for example 100%, of the top periphery (302) of the through-hole must be within the distance (d) of at least one conductive point (121) of the grid electrode (120), wherein the top periphery (302) is coplanar with the surface of the grid electrode (120) attached to the grid (112). This embodiment ensures that a large portion of the electrode is very close to the through-hole, thereby improving measurement accuracy. Figure 3 At least 75% of the top periphery (302) is within the distance (d).

[0061] In an embodiment, at least two of the at least one grid electrodes can be electrically connected via a switch (401), wherein the at least two grid electrodes are located within the distance from the same through-hole. When the switch (401) is turned on, the connection between the two electrodes is closed, and the two electrodes act as a single electrode. This is in Figure 4 This is described in the description. This allows for electrical measurements of the cell barrier at the pore by measuring the signal between an electrode connected at one end and a capping electrode at the other end. Because the connected electrode surrounds more of the pore than each electrode individually, a larger signal is obtained, thus exhibiting higher sensitivity. When the switch (401) is turned off, the connection between the two electrodes is broken, and the two electrodes can operate independently. This allows for electrical measurements of the cell barrier at two different locations around the pore by measuring the signal between each electrode connected at one end and a capping electrode at the other end. This also allows for lateral measurements between the two electrodes without involving the capping electrode, thereby measuring what is happening between the electrodes (e.g., the presence of a chemical compound).

[0062] As used herein, unless otherwise stated, the term "switch" refers to a device for establishing or disconnecting an electrical connection. In the case of a semiconductor cell culture device, the switch (401) is used to electrically connect two or more grid electrodes having at least one conductive point located at a distance from the same via.

[0063] In an embodiment, the geometry may be further specified as follows: the via must have a top periphery coplanar with the surface of at least one grid electrode attached to the grid, the top periphery having an average diameter (D) such that the ratio between the average diameter and the distance (d) separating the at least one conductive point (121) from the via is at least 2, preferably at least 3, more preferably at least 5, and most preferably at least 9. Figure 3 In this case, the ratio is 9. The higher the ratio, the better the information collected by the grid electrodes reflects the condition of the cell barrier at the level of the pores (114).

[0064] In this embodiment, the first and second electrical connections allow for simultaneous electrical measurements between at least two of a group of one or more of the same cap electrode and at least one of the at least one grid electrode, each electrode in the group satisfying the geometry with respect to the same via. This embodiment allows for simultaneous measurements at different grid electrodes around the same via, providing a more comprehensive understanding of the electrical properties of cell cultures directly above the via (114).

[0065] In an embodiment, the group may be a group of all grid electrodes among the at least one grid electrode that satisfy the geometric conditions with respect to the same via.

[0066] In an embodiment, each through-hole may have a top periphery, and each of the at least one cap electrode is directly located on: a) a different through-hole such that the vertical projection of the electrode on the plane of the top surface of the grid at least partially overlaps the through-hole, or b) a plurality of through-holes such that the vertical projection of the electrode on the plane of the top surface of the grid completely includes the top periphery of the plurality of through-holes. Figure 8 Scenario b is described in the text, while Figure 14 The cap electrode is depicted in (right).

[0067] In an embodiment, the at least one cap electrode (see, for example) Figure 13 and 14 Each of the electrodes can have an opening and can be positioned directly above the plurality of through-holes such that the vertical projection of the electrode onto the plane of the top surface of the grid completely encompasses the top periphery of the plurality of through-holes and excludes at least some space between two adjacent through-holes. This situation is as follows: Figure 9 As shown. These openings allow observation of cell cultures remotely from the wells (e.g., via spectroscopy or microscopy).

[0068] In an embodiment, each of the at least one cap electrode may have an opening and may be positioned such that the vertical projection of the opening onto the plane of the top surface of the mesh completely encompasses the top periphery of the plurality of through holes. This is as follows: Figure 10 As shown. These openings allow observation of the cell culture above the wells. Figure 9 and 10 The embodiments can also be combined such that the vertical projection of some openings does not overlap with the through hole, while the vertical projection of other openings overlaps with the through hole.

[0069] In an embodiment, one or more of the at least one cap electrode may have the same shape, and the through-hole may have the same shape. This is in Figure 16 As shown in the figure, Figure 16 This is a schematic top view of the grid electrodes surrounding the hexagonal through-hole (left) and the hexagonal cap electrode (right) used in an embodiment of the present invention.

[0070] We now refer to Figure 11 . Figure 11 This is a schematic representation of a semiconductor cell culture apparatus, showing electrodes on the bottom surface of a grid, a first electrical connection (110) and a second electrical connection (111) to the grid electrode (120) and the cap electrode (220), respectively, an electrical measurement system (117) connected to the first electrical connection (110) and the second electrical connection (111), and a processing unit (119) for performing the method according to the second aspect of the invention.

[0071] Therefore, in an embodiment, the semiconductor cell culture apparatus may further include an electrical measurement system (117) connected to the first and second electrical connections, which is configured to perform measurements between any of the at least one grid electrode (120) and the cap electrode (220).

[0072] Figure 12 This is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture apparatus according to an embodiment of the present invention, showing a cap facing the top surface of a semiconductor grid and defining a space for cell cultures, and having multiple electrodes on the bottom surface of the cap and multiple electrodes on the bottom surface of the grid, each electrode being at a different distance from a via. This allows for measurements to be performed in a four-electrode configuration (two grid electrodes and two cap electrodes), wherein a current is applied between a grid electrode (123) and an inner cap electrode (220i), the grid electrode (123) being farther from the via than the electrode (120), and voltage can be measured between the grid electrode (120) and the inner cap electrode (220i) at a distance greater than d from the via, and closer to the via than the electrode (123) and within said distance d.

[0073] Figure 13 and 14 This is a schematic top view of an electrode for attachment to the bottom surface of a cover according to an embodiment of the present invention. Figure 13 It has an opening that allows observation (e.g., spectroscopy or microscopy). Figure 14 On the left is a grid electrode, which also allows for observation. Figure 14 (Right) is available for use Figure 8 The planar electrode in the embodiment.

[0074] Figure 15 This is a schematic top view of the grid electrodes used in embodiments of the present invention. Two different shapes are shown: circular and hexagonal. The electrode (120) on the left is shown as the periphery surrounding the through-hole (302) and can be used in a two-electrode configuration. The electrodes on the right are a first electrode within the distance d and a second electrode that can be further away than the distance d. These two electrodes (120, 123) can be used in a four-electrode configuration.

[0075] Figure 16 This is a schematic representation of a detailed top view of the grid electrode (left) and the cap electrode (right) used in an embodiment of the present invention.

[0076] We now refer to Figure 17In a second aspect, the present invention relates to a method for detecting the presence of defects in a biological barrier of a semiconductor cell culture apparatus according to any embodiment of the first aspect. The method comprises sequentially initiating electrical measurements between each grid electrode and cap electrode (220) of the apparatus, generating an electrical output, and determining from the output that if any grid electrode measures an output outside a predetermined range, it indicates the presence of a defect in the cell culture (204) above a via (114) within the distance between these electrodes.

[0077] In this embodiment, the electrical measurement is an electrochemical measurement. For example, it could be a transepithelial-transendothelial electrical measurement. This can be performed using methods such as impedance spectroscopy or other electrical measurement techniques.

[0078] As used herein, unless otherwise stated, the term "transepithelial-transendothelial electrometry" refers to an electrical measurement performed across epithelial or endothelial cell layers. This measurement is used to assess the integrity and function of these cell layers.

[0079] As used herein, unless otherwise stated, the term "impedance spectroscopy measurement" refers to an electrical measurement used to determine the impedance or resistance to current of a material over a range of frequencies. This measurement can be used in the study of biological cells and tissues.

[0080] As used herein, unless otherwise stated, the term "electrical measurement system" refers to a system connected to the first and second electrical connections (110, 111) and configured to perform measurements between any grid electrode and cover electrode (220). This system may include various components such as power supplies, instruments, and other electronic equipment.

[0081] Any feature of the second aspect can be described accordingly in any other aspect.

[0082] In a third aspect, the present invention relates to a semiconductor cell culture system comprising means according to any embodiment of the first aspect, and further comprising a processing unit (119) adapted to perform the method of the second aspect.

[0083] As used herein, unless otherwise stated, the term "processing unit" refers to a component of a semiconductor cell culture system adapted to perform methods for detecting the presence of defects in biological barriers. This unit may include a computer or other electronic device capable of processing data and executing instructions.

[0084] Any feature of the third aspect can be described accordingly in any other aspect.

[0085] In a fourth aspect, the present invention relates to a computer program comprising instructions for causing a semiconductor cell culture system of the third aspect to perform the steps of the method of the second aspect.

[0086] Any feature of the fourth aspect may be described accordingly in any other aspect.

[0087] In a fifth aspect, the present invention relates to a computer-readable medium having a computer program of the fourth aspect stored thereon.

[0088] Any feature of the fifth aspect may be described accordingly in any other aspect.

[0089] It is understood that although preferred embodiments, specific structures and configurations, and materials have been discussed herein with respect to the device according to the invention, various changes or modifications in form and detail may be made without departing from the scope of the invention. For example, any molecular formulas given above represent only steps that can be used. Functions can be added or removed from the block diagrams, and operations can be interchanged between function blocks. Steps can be added or removed from the method within the scope of the invention.

Claims

1. A semiconductor cell culture device, comprising: a. A semiconductor mesh (112) having islands (116) interconnected by bridges (118) and defining vias (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface. b. A cap (200), facing and separated from the top surface of the semiconductor grid (112), thereby defining a space (202) for cell culture (204), wherein the cap (200) has a top surface and a bottom surface. c. At least one grid electrode (120) attached to the top or bottom surface of the grid. d. At least one cover electrode (220) attached to the bottom surface of the cover (200). e. A first electrical connection (110) to one or more of the at least one grid electrode (120), and f. A second electrical connection (111) to one or more of the at least one cap electrode (220). The first electrical connection and the second electrical connection (111) allow electrical measurements to be performed between each of the at least one or more of the grid electrodes (120) and the cap electrode (220), and The at least one or more of the grid electrodes (120) satisfy a geometric condition that specifies that at least one conductive point (121) of each of the at least one or more of the grid electrodes (120) must be located within a distance (d) of 100 μm from the via (114).

2. The semiconductor cell culture apparatus according to claim 1, characterized in that, The distance (d) from the through hole (114) is 10µm, preferably 5µm.

3. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, The geometric conditions further specify that at least 50%, preferably at least 75%, more preferably at least 90%, for example 100%, of the top periphery (302) of the through hole (114) must be within the distance (d) of at least one conductive point of the at least one grid electrode (120), and the top periphery (302) must be coplanar with the surface of the at least one grid electrode (120) attached to the grid.

4. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, The geometric conditions further specify that the through hole (114) must have a top periphery (302) coplanar with the surface of the at least one grid electrode (120) attached to the grid, the top periphery (302) having an average diameter such that the ratio of the average diameter (D) to the distance (d) separating the at least one conductive point (121) from the through hole is at least 2, preferably at least 3, and more preferably at least 5.

5. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, The first electrical connection and the second electrical connection (111) allow simultaneous electrical measurements between at least two of a group of one or more of the same cap electrode (220) and the at least one grid electrode (120), each electrode of the group satisfying the geometry with respect to the same via (114).

6. The semiconductor cell culture apparatus according to claim 5, characterized in that, The group is a group of all grid electrodes among the at least one grid electrode (120) that satisfy the geometric conditions with respect to the same through hole (114).

7. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, Each through-hole has a top periphery (302), and one or more of said at least one cap electrode (220) are directly above the following: a. A through-hole, such that the vertical projection of the electrode onto the plane of the top surface of the grid at least partially overlaps with the through-hole, or b. Multiple through holes such that the vertical projection of the electrode onto the plane of the top surface of the grid completely includes the top periphery (302) of the multiple through holes.

8. The semiconductor cell culture apparatus according to claim 7, characterized in that, Each of the at least one cap electrode (220) is located directly above the plurality of through holes, such that the vertical projection of the electrode onto the plane of the top surface of the grid completely includes the top periphery (302) of the plurality of through holes and does not include at least some space between two adjacent through holes.

9. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, One or more of the at least one cap electrode (220) have the same shape, and the through hole has the same shape.

10. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, It also includes an electrical measurement system (117) connected to the first electrical connection and the second electrical connection (111), the electrical measurement system (117) being configured to perform measurements between any one of the at least one grid electrode (120) and the cover electrode (220).

11. The semiconductor cell culture apparatus according to any one of the preceding claims, characterized in that, The electrical measurements are selected from transepithelial-transendothelial electrical measurements and impedance spectroscopy measurements.

12. A method for detecting the presence of defects in a biological barrier in a semiconductor cell culture apparatus according to any one of the preceding claims, the method comprising: a. An electrical measurement is sequentially initiated between each of one or more of the at least one grid electrode (120) and the cap electrode (220) of the semiconductor cell culture apparatus, thereby generating an electrical output. b. Determine, based on the electrical output obtained from the electrical measurement: if any one or more of the at least one grid electrode (120) measures an output outside a predetermined range, it indicates a defect in the cell culture (204) above the through-hole (114) within the distance (d) of these electrodes.

13. A semiconductor cell culture system comprising the apparatus according to any one of claims 1 to 11, and further comprising a processing unit (119) adapted to perform the method of claim 12.

14. A computer program comprising instructions for causing the semiconductor cell culture system of claim 13 to perform the steps of the method of claim 12.

15. A computer-readable storage medium having thereon a computer program as claimed in claim 14.

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