WO2025119966A1

WO2025119966A1 - Semiconductor mesh for cell barrier analysis

Info

Publication number: WO2025119966A1
Application number: PCT/EP2024/084653
Authority: WO
Inventors: Saeedeh EBRAHIMI TAKALLOO; Dries Braeken; Maria del Mar CONDOR
Original assignee: Interuniversitair Microelektronica Centrum vzw IMEC
Current assignee: Interuniversitair Microelektronica Centrum vzw IMEC
Priority date: 2023-12-05
Filing date: 2024-12-04
Publication date: 2025-06-12
Anticipated expiration: 2026-06-05

Abstract

Semiconductor Mesh for Cell Barrier Analysis The patent application pertains to a semiconductor cell culture device The device includes a semiconductor mesh with interconnected islands and through-pores, a cover defining a cell culture space, mesh and cover electrodes, and electrical connections. The electrical connections allow for electrical measurements between the mesh and cover electrodes. The mesh electrodes must satisfy a geometrical condition that at least one electrically conductive point must be located within a specific distance from a through-pore. The device allows for the detection of faults in a biological barrier, with the electrical measurements providing outputs indicative of any faults. The device can be part of a system that includes a processing unit adapted to perform the method of fault detection.

Description

Semiconductor Mesh for Cell Barrier Analysis

Field of the Invention

The present invention relates to the field of biomedical engineering and cell culture devices, and more specifically to a semiconductor cell culture device and method for detecting faults in a biological barrier.

Background of the Invention

The field of cell culture and barrier integrity analysis is a critical area of research in biomedical science. This field is particularly relevant to the study of biological barriers such as the blood-brain barrier (BBB), gut barrier, lung barrier, liver barrier, and others. These barriers play a crucial role in protecting the body from harmful substances, and their integrity is essential for maintaining health.

One of the primary methods for studying these barriers and their integrity is through the use of cell culture devices. These devices often incorporate a mesh structure that serves as a foundation for cell layer growth. The mesh structure typically includes through-pores, which are spaces between the mesh islands where cells can grow and form a barrier.

A significant challenge in this field is the accurate measurement of barrier integrity, such as the detection of micro-level breaches in the barrier. Traditional methods, such as transepithelial-transendothelial electrical 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 electrical resistance between them. However, these methods have limitations. For instance, they may not provide a detailed, localized assessment of barrier integrity at the level of individual pores. Also, traditional methods may not be sensitive enough to detect small breaches, which can have significant implications for barrier function.

Despite the advancements in cell culture devices and methods for barrier integrity analysis, there are still significant challenges in this field. There is a need for further advancements to address these challenges and improve the accuracy and sensitivity of barrier integrity measurements. Summary of the Invention

It is an object of embodiments of the present invention to provide devices and method capable of good barrier integrity analysis, particularly for the blood-brain barrier, by enabling micro-level breach detection. This objective is accomplished by a semiconductor cell culture device, a system, and a method according to the invention.

In the first aspect, the present invention relates to a semiconductor cell culture device comprising a semiconductor mesh with interconnected islands and through-pores, a cover facing the mesh and defining a space for cell culture, at least one mesh electrode attached to the mesh, at least one cover electrode attached to the cover, first and second electrical connections (111) to the mesh and cover electrodes respectively, allowing for electrical measurement between the electrodes. The mesh electrode must satisfy a geometrical condition that at least one electrically conductive point must be located within a distance of 100 pm from a through-pore. 100 pm is the maximum endothelia l/epithelial cell radius in culture.

Preferably, it relates to a semiconductor cell culture device comprising: a. a semiconductor mesh (112) having islands (116) being interconnected by bridges (118) and defining through-pores (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface, b. a cover facing the top surface of the semiconductor mesh (112) and separated therefrom, thereby defining a space for a cell culture, wherein the cover has a top surface and a bottom surface, c. at least one mesh electrode attached to the top surface or the bottom surface of the mesh, d. at least one cover electrode attached to the bottom surface of the cover, e. first electrical connections (110) to one or more of the at least one mesh electrode, and f. second electrical connections (111) to one or more of the at least one cover electrode, wherein the first and second electrical connections allow the performance of an electrical measurement between each of said one or more of the at least one mesh electrode with a cover electrode, and wherein said one or more of the at least one mesh electrode satisfy a geometrical condition imposing that at least one electrically conductive point of each of said one or more of the at least one mesh electrode must be located within a distance (d) of 100 pm from a through-pore (114).

In the second aspect, the present invention relates to a method for detecting the presence of a fault in a biological barrier in a semiconductor cell culture device according to any embodiments of the first aspect. The method comprises initiating an electrical measurement sequentially between each of the mesh electrodes and a cover electrode of the device, generating electrical outputs, and determining from the outputs which, if any of the mesh electrodes measure an output outside a predetermined value range, indicative of the presence of a fault in the cell culture above a through-pore within said distance of these electrodes.

Preferably, it may relate to a method for detecting the presence of a fault in a biological barrier in a semiconductor cell culture device according to any one of the preceding claims, the method comprising: a. Initiating an electrical measurement sequentially between each of the one or more of the at least one mesh electrode and a cover electrode of the semiconductor cell culture device, thereby generating electrical outputs, b. Determining from the electrical outputs obtained from the electrical measurements which, if any of the one or more of the at least one mesh electrode measure an output outside a predetermined value range, indicative of the presence of a fault in the cell culture above a through-pore (114) within said distance (d) of these electrodes.

In the third aspect, the present invention relates to a semiconductor cell culture system comprising a device according to any embodiments of the first aspect, and further comprising a processing unit adapted to perform the method of the second aspect.

In the fourth aspect, the present invention relates to a computer program comprising instructions to cause the semiconductor cell culture system of the third aspect to execute the steps of the method of the second aspect. In the fifth aspect, the present invention relates to a computer-readable medium having stored thereon the computer program of the fourth aspect.

It is an advantage of embodiments of the present invention that a semiconductor cell culture device can be achieved, which allows for precise and accurate measurements of barrier integrity at a micro-level. It is a further advantage of embodiments of the present invention that the device can discriminate pore defects, enhancing the accuracy of breach detection.

It is a further advantage of embodiments of the present invention that the device can detect faults such as holes, irregularities, disruptions, heterogeneities, or tightness inconsistencies in the barrier, in particular at the level of pores. This feature is particularly useful in drug delivery and barrier integrity analysis, as it allows for clear observation and electrical measurements for barrier integrity.

It is a further advantage of embodiments of the present invention that the device can be integrated into microfluidic chips. This integration allows for a more streamlined and efficient process, saving time and resources for researchers and scientists.

Particular and preferred aspects of the invention are set out in the accompanying independent and dependent claims. Features from the dependent claims may be combined with features of the independent claims and with features of other dependent claims as appropriate and not merely as explicitly set out in the claims.

The above and other characteristics, features and advantages of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of the invention. This description is given for the sake of example only, without limiting the scope of the invention. The reference figures quoted below refer to the attached drawings.

Brief description of the drawings

Fig.l is a schematic representation of a top view of horizontal cross-section through the semiconductor cell culture device, showing the semiconductor mesh with islands interconnected by bridges and defining through-pores, according to the prior art. Fig.2 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having electrodes on the cover and the bottom for allowing TEER according to the prior art.

Fig.3 is a detailed top view of a schematic representation of part of the semiconductor cell culture device, illustrating the geometrical condition of the mesh electrode in relation to the through-pore, according to embodiments of the present invention.

Fig.4 is a schematic representation of the semiconductor cell culture device, showing the electrical connection between two mesh electrodes through a switch, according to embodiments of the present invention.

Fig.5 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having electrodes on the bottom surface of the cover and the top of the mesh for allowing electrical measurements according to embodiments of the present invention.

Fig.6 illustrates a leak due to a fault in the cell culture above through-hole in the embodiments of Fig.5.

Fig. 7 illustrates the path taken by electric charges during an electrical measurement in a device analogous to the one illustrated in Figs. 6 and 7.

Fig. 8 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having a single large electrode on the bottom surface of the cover and multiple electrodes on the top of the mesh for allowing electrical measurements according to embodiments of the present invention.

Figs. 9 and 10 are schematic representations of a vertical cross-sectional views of semiconductor cell culture devices, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having a single large electrode pierced by holes on the bottom surface of the cover and multiple electrodes on the top of the mesh for allowing electrical measurements according to embodiments of the present invention and for allowing observation of the cells (e.g., spectroscopy or microscopy).

Fig.11 is a schematic representation of the semiconductor cell culture device, demonstrating electrodes on the bottom surface of the mesh, the first and second electrical connections to the mesh and cover electrodes respectively, the electrical measurement system connected to the first and second electrical connections, as well as the processing unit for performing the method according to embodiments of the present invention.

Fig. 12 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having a plurality of electrodes on the bottom surface of the cover and multiple electrodes on the bottom surface of the mesh, at different distances from a through-hole, according to embodiments of the present invention.

Figs. 13 and 14 are schematic representation of detailed top views of electrodes for attachment to the bottom surface of the cover, according to embodiments of the present invention.

Fig. 15 is a schematic representation of a detailed top view of mesh electrodes for use in embodiments of the present invention.

Fig. 16 is a schematic representation of a detailed top view of mesh electrodes (left) and cover electrodes (right) for use in embodiments of the present invention.

Fig.17 is a flowchart illustrating the method for detecting the presence of a fault in a biological barrier in the semiconductor cell culture device, according to embodiments of the present invention.

In the different figures, the same reference signs refer to the same or analogous elements.

Detailed description of Illustrative Embodiments

The present invention will be described with respect to particular embodiments and with reference to certain drawings but the invention is not limited thereto but only by the claims. The drawings described are only schematic and are non-limiting. In the drawings, the size of some of the elements may be exaggerated and not drawn on scale for illustrative purposes. The dimensions and the relative dimensions do not correspond to actual reductions to practice of the invention.

The terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequence, either temporally, spatially, in ranking or in any other manner. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.

Moreover, the terms top and over and the like in the description and the claims are used for descriptive purposes and not necessarily for describing relative positions. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other orientations than described or illustrated herein.

It is to be noticed that the term "comprising", used in the claims, should not be interpreted as being restricted to the means listed thereafter; it does not exclude other elements or steps. It is thus to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. The term "comprising" therefore covers the situation where only the stated features are present and the situation where these features and one or more other features are present. The word "comprising" according to the invention therefore also includes as one embodiment that no further components are present. Thus, the scope of the expression "a device comprising means A and B" should not be interpreted as being limited to devices consisting only of components A and B. It means that with respect to the present invention, the only relevant components of the device are A and B.

Similarly, it is to be noticed that the term "coupled" should not be interpreted as being restricted to direct connections only. The terms "coupled" and "connected", along with their derivatives, may be used. It should be understood that these terms are not intended as synonyms for each other. Thus, the scope of the expression "a device A coupled to a device B" should not be limited to devices or systems wherein an output of device A is directly connected to an input of device B. It means that there exists a path between an output of A and an input of B which may be a path including other devices or means. "Coupled" may mean that two or more elements are either in direct physical or electrical contact, or that two or more elements are not in direct contact with each other but yet still co-operate or interact with each other.

Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment, but may. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as would be apparent to one of ordinary skill in the art from this disclosure, in one or more embodiments.

Similarly it should be appreciated that in the description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding in the understanding of one or more of the various inventive aspects. This method of disclosure, however, is not to be interpreted as reflecting an intention that the claimed invention requires more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single foregoing disclosed embodiment. Thus, the claims following the detailed description are hereby expressly incorporated into this detailed description, with each claim standing on its own as a separate embodiment of this invention.

Furthermore, while some embodiments described herein include some but not other features included in other embodiments, combinations of features of different embodiments are meant to be within the scope of the invention, and form different embodiments, as would be understood by those in the art. For example, in the following claims, any of the claimed embodiments can be used in any combination.

Furthermore, some of the embodiments are described herein as a method or combination of elements of a method that can be implemented by a processor of a computer system or by other means of carrying out the function. Thus, a processor with the necessary instructions for carrying out such a method or element of a method forms a means for carrying out the method or element of a method. Furthermore, an element described herein of an apparatus embodiment is an example of a means for carrying out the function performed by the element for the purpose of carrying out the invention.

In the description provided herein, numerous specific details are set forth. However, it is understood that embodiments of the invention may be practiced without these specific details. In other instances, well-known methods, structures and techniques have not been shown in detail in order not to obscure an understanding of this description.

The following terms are provided solely to aid in the understanding of the invention.

The invention will now be described by a detailed description of several embodiments of the invention. It is clear that other embodiments of the invention can be configured according to the knowledge of persons skilled in the art without departing from the technical teaching of the invention, the invention being limited only by the terms of the appended claims.

We now refer to Fig. 1. Fig.l is a schematic representation of a top view of horizontal cross-section through the semiconductor cell culture device, showing the semiconductor mesh (112) with islands (116) interconnected by bridges (118) and defining through-pores (114), according to the prior art. A mesh electrode (120) is depicted at the center of an island (116), far from the edge of any through pore.

We now refer to Fig. 2. Fig.2 is a schematic representation of a vertical cross- sectional view of a semiconductor cell culture device according to the prior art. It shows the cover (200) facing the top surface of the semiconductor mesh (112) and defining a space (202) for a cell culture (204), and having cover electrodes (220) on the cover (200) and bottom electrodes (230) on the bottom of the device for allowing TEER according to the prior art. A cell culture (204) is depicted on the mesh (112).

The device of the present invention permits to discriminate faults in a biological barrier, such as a cell culture, at the level of the through-pore, i.e., where faults are most critical.

Fig.5 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device according to the first aspect. As used herein, and unless otherwise specified, the term "semiconductor cell culture device" refers to a device that is used for the growth and maintenance of biological cells. This device comprise semiconductor materials. Fig. 5 illustrates the cover (200) facing the top surface of the semiconductor mesh (112) and separated therefrom, thereby defining a space (202) for a 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 device that faces the top surface of the semiconductor mesh and is separated from it. This cover (200) also has a top surface and a bottom surface, and it defines a space (202) for cell culture (204) when positioned over the semiconductor mesh.

The device further has at least one cover electrode (220) attached to the bottom surface of the cover (200) and at least one mesh electrode (120) attached to the top surface of the mesh (112) for allowing electrical measurements according to embodiments of the present invention. The cover electrodes are electrodes that are attached to the bottom surface of the cover (200). These electrodes are used for conducting electrical measurements in the semiconductor cell culture device. The mesh electrodes are also used for conducting electrical measurements in the semiconductor cell culture device. The mesh electrodes can also be attached to the bottom of the mesh (see Figs. 11 and 12). The one or more of the at least one mesh electrode (120) satisfy a geometrical condition imposing that at least one electrically conductive point (121) of each of said one or more of the at least one mesh electrode (120) must be located within a distance (d) of 100 pm from a through-pore (114).

The semiconductor mesh (112) has islands (116) being interconnected by bridges (118) (not depicted) and defining through-pores (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface. In other words, the semiconductor mesh refers to a network or fabric-like structure made from semiconductor material. This mesh includes interconnected islands and bridges, and defines through-pores between the islands. The mesh has a top surface and a bottom surface. The islands, bridges, and through- pores are all integral parts of the mesh structure. First electrical connections (110) to one or more of the at least one mesh electrode (120), and second electrical connections to one or more of the at least one cover electrode (220) are present but not depicted in Fig. 5. The first and second electrical connections allow the performance of an electrical measurement between each of said one or more of the at least one mesh electrode (120) with a cover electrode (220). The first electrical connections (110) are connections that are made to one or more of the mesh electrodes. These connections allow for the transmission of electrical signals between the mesh electrodes and other components of the semiconductor cell culture device. The second connections are connections that are made to one or more of the cover electrodes. These connections allow for the transmission of electrical signals between the cover electrodes and other components of the semiconductor cell culture device. An electrical measurement is depicted in Fig. 7. The electrical measurement is a process of determining the electrical properties of the cell culture (204). This measurement is performed, typically sequentially, between each of the one or more of the at least one mesh electrodes and a cover electrode (220), and it is facilitated by the first and second electrical connections. Such an electrical measurement permits to detect a fault in the cell culture at the level of a through-pore (114). Such a fault is depicted in fig. 6 where chemical species supposed to be confined in the space (202) have passed through a fault in the cell culture (204) at the level of a through-pore (114).

Fig.3 is a detailed top view of a schematic representation of part of the semiconductor cell culture device, illustrating the geometrical condition of the mesh electrode (120) in relation to the through-pore (114), according to embodiments of the present invention. As used herein, and unless otherwise specified, the term "geometrical condition" refers to a specific spatial requirement that must be met by the mesh electrodes. This condition stipulates that at least one electrically conductive point (121) of one or more of the at least one mesh electrode (120) must be located within a certain distance (d) from a through-pore (114).

In embodiments, the distance may be 10 pm, preferably 5 pm from a through-pore. This embodiment allows for more precise measurements due to the closer proximity of the electrode to the through-pore. Most preferably, said distance may be 1 pm, or even 0 pm (e.g., one or more of the at least one electrically conductive point from the first electrode and one or more of the at least one electrically conductive point from the second electrode must be in direct contact or immediate proximity with said single through-pore so that there is substantially no distance between them and the through-pore). In embodiments, the geometrical condition may impose that each mesh electrode (120) must have an electrically conductive boundary with at least two distinct conductive points located within the distance and equidistant from the through-pore. This is illustrated in Fig. 3. This embodiment ensures uniformity in the measurements.

In embodiments, each mesh electrode (120) may have an electrically conductive boundary of its top surface having a line (122) being at least 20 pm long and entirely located within the distance from the through-pore (114). This is also illustrated in Fig. 3. The line is encircled by a dashed rectangle. This embodiment allows for a larger area of the electrode to be in close proximity to the through-pore, enhancing the accuracy of the measurements. In embodiments, the line may run parallel to a periphery of the through-pore, the periphery being coplanar with the surface of the mesh electrode (120) attached to the mesh. This is also the case in Fig. 3. This embodiment ensures that the electrode is in the optimal position for accurate measurements.

In embodiments, the geometrical condition may further impose that at least 50%, preferably at least 75%, more preferably at least 90%, such as 100% of the top periphery (302) of the through-pore, the top periphery (302) being coplanar with that surface of the mesh electrode (120) which is attached to the mesh (112), must be within said distance (d) of at least one electrically conductive point (121) of the mesh electrode (120). This embodiment ensures that a significant portion of the electrode is in close proximity to the through-pore, enhancing the accuracy of the measurements. In Fig. 3, at least 75% of the top periphery (302) is within said distance (d)

In embodiments, at least two of said one or more of the at least one mesh electrode, both being located within said distance from a same through-pore, may be electrically connected through a switch (401). When the switch (401) is on, the connection between both electrodes is closed and both electrodes act as a single electrode. This is depicted in Fig. 4. This permits an electrical measurement of the cell barrier at that pore by measuring a signal between the connected electrodes on one hand and the cover electrode on another hand. Since the connected electrode surround more of the through pore than each electrode alone the signal obtained is larger and therefor exhibits a higher sensitivity. When the switch (401) is off, the connection between both electrodes is open and both electrodes can act separately. This permits each electrode to perform an electrical measurement of the cell barrier at two different locations around the pore by measuring a signal between the each of the separated electrodes on one hand and the cover electrode on another hand. This also permits to perform measurement laterally between both electrodes, without involving the cover electrode, thereby measuring what happen between the electrodes (e.g., the presence of chemical compounds).

As used herein, and unless otherwise specified, the term "switch" refers to a device that is used to make or break an electrical connection. In the context of the semiconductor cell culture device, a switch (401) is used to electrically connect two or more mesh electrodes that have at least one electrically conductive point located within a certain distance from the same through-pore. In embodiments, the geometrical condition may further impose that said through- pore must have a top periphery being coplanar with that surface of the at least one mesh electrode which is attached to the mesh, said top periphery having an average diameter (D) such that the ratio between said average diameter and the distance (d) separating said at least one electrically conductive point (121) from the through-pore is at least 2, preferably at least 3, more preferably at least 5, and most preferably at least 9. In the case of Fig. 3, this ratio is 9. The higher this ratio, the better the information gathered by the mesh electrode reflect the situation of the cell barrier at the level of the through-pore (114).

In embodiments, the first and second electrical connections may allow the simultaneous performance of an electrical measurement between a same cover electrode and a group of at least two of said one or more of the at least one mesh electrode, each electrode of said group satisfying said geometrical condition with respect to a same through- pore. This embodiment allows for simultaneous measurements at various mesh electrodes around a same through pore, providing a more comprehensive understanding of the electrical properties of the cell culture directly above that through-pore (114).

In embodiments, said group may be a group of all of said one or more of the at least one mesh electrode satisfying said geometrical condition with respect to said same through- pore.

In embodiments, each through-pore may have a top periphery, and wherein each of the at least one cover electrode is positioned directly above: a. a different through-pore such that a vertical projection of the electrode on the plane of the top surface of the mesh at least partially overlap with the through-pore, or b. a plurality of through-pores such that a vertical projection of the electrode on the plane of the top surface of the mesh entirely comprises the top periphery of the plurality of through-pores. Situation b. is depicted in Fig. 8 and the cover electrode is depicted in Fig. 14 (right).

In embodiments, each of the at least one cover electrode (see e.g., Figs. 13 and 14) may have openings and may be positioned directly above a plurality of through-pores such that a vertical projection of the electrode on the plane of the top surface of the mesh entirely comprises the top periphery of the plurality of through-pores and does not comprise at least some space between two adjacent through-pores. This situation is depicted in Fig. 9. These openings permit the observation of the cell culture away from the pores (e.g., by spectroscopy or microscopy). In embodiments, each of the at least one cover electrode may have openings and may be positioned such that a vertical projection of the openings on the plane of the top surface of the mesh entirely comprises the top periphery of the plurality of through-pores. This situation is depicted in Fig. 10. These openings permit the observation of the cell culture above the pores. The embodiments of Fig. 9 and 10 can also be combined so that the vertical projection of some openings is not overlapping with a through hole while the vertical projection of other openings do overlap with a through hole.

In embodiments, the one or more of the at least one cover electrode may have a same shape and wherein the through-pores have said same shape. This is depicted in Fig. 16, which is a schematic representation of a detailed top view of mesh electrodes around hexagonal through-holes (left) and hexagonal cover electrodes (right) for use in embodiments of the present invention.

We now refer to Fig. 11. Fig.11 is a schematic representation of the semiconductor cell culture device, demonstrating electrodes on the bottom surface of the mesh, the first (110) and second (111) electrical connections to the mesh (120) and cover (220) electrodes respectively, the electrical measurement system (117) connected to the first (110) and second (111) electrical connections, as well as the processing unit (119) for performing the method according to the second aspect of the present invention.

Hence, in embodiments, the semiconductor cell culture device may further comprise an electrical measurement system (117) connected to the first and second electrical connections, configured to perform measurements between any of the at least one mesh electrode (120) and a cover electrode (220).

Fig. 12 is a schematic representation of a vertical cross-sectional view of a semiconductor cell culture device, illustrating the cover facing the top surface of the semiconductor mesh and defining a space for a cell culture, and having a plurality of electrodes on the bottom surface of the cover and multiple electrodes on the bottom surface of the mesh, at different distances from a through-hole, according to embodiments of the present invention. This permit the performance of measurements in a four electrodes configuration (two mesh electrodes and two cover electrodes) wherein the current is applied between the mesh electrode (123), which is farther from the through-hole than electrode (120) and which can be at a distance larger than d from the through hole, and the inner cover electrode (220i), and the voltage is measured between the mesh electrode (120) which is closer to the through-hole than electrode (123) and which is within said distance d, and the inner cover electrode (220i).

Figs. 13 and 14 are schematic representation of detailed top views of electrodes for attachment to the bottom surface of the cover, according to embodiments of the present invention. Fig. 13 has openings allowing observation (e.g., spectroscopy or microscopy). Fig. 14 left is a grid electrode, which also allows observation. Fig. 14 (right) is a plain electrode as can be used in the embodiment of Fig. 8.

Fig. 15 is a schematic representation of a detailed top view of mesh electrodes for use in embodiments of the present invention. Two different shapes are represented: circular and hexagonal. The electrode (120) on the left is shown around the periphery of a through hole (302) and can be used in a two electrode configuration. The electrodes on the right are a first electrode within said distance d and a second electrode which can be farther than said distance d. These two electrodes (120, 123) can be used in a four electrodes configuration.

We now refer to Fig. 17. In the second aspect, the present invention relates to a method for detecting the presence of a fault in a biological barrier in a semiconductor cell culture device according to any embodiments of the first aspect. The method comprises initiating an electrical measurement sequentially between each of the mesh electrodes and a cover electrode (220) of the device, generating electrical outputs, and determining from the outputs which, if any of the mesh electrodes measure an output outside a predetermined value range, indicative of the presence of a fault in the cell culture (204) above a through-pore (114) within the distance of these electrodes.

In embodiments, the electrical measurement is an electrochemical measurement. For instance, it may be a transepithelial-transendothelial electrical measurement. This can be conducted using methods such as impedance spectroscopy measurement or other electrical measurement techniques.

As used herein, and unless otherwise specified, the term "transepithelial- transendothelial electrical measurement" refers to a type of electrical measurement that is performed across epithelial or endothelial cell layers. This measurement is used to assess the integrity and function of these cell layers. As used herein, and unless otherwise specified, the term "impedance spectroscopy measurement" refers to a type of electrical measurement that is used to determine the impedance, or resistance to electrical flow, of a material over a range of frequencies. This measurement can be used in the study of biological cells and tissues.

As used herein, and unless otherwise specified, the term "electrical measurement system" refers to a system that is connected to the first and second electrical connections (110, 111) and is configured to perform measurements between any of the mesh electrodes and a cover electrode (220). This system may include various components such as power sources, meters, and other electronic devices.

Any feature of the second aspect can be as correspondingly described in any of the other aspects.

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

As used herein, and unless otherwise specified, the term "processing unit" refers to a component of the semiconductor cell culture system that is adapted to perform a method for detecting the presence of a fault in a biological barrier. This unit may include a computer or other electronic device that is capable of processing data and executing instructions.

Any feature of the third aspect can be as correspondingly described in any of the other aspects.

In the fourth aspect, the present invention relates to a computer program comprising instructions to cause the semiconductor cell culture system of the third aspect to execute the steps of the method of the second aspect.

Any feature of the fourth aspect can be as correspondingly described in any of the other aspects.

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

Any feature of the fifth aspect can be as correspondingly described in any of the other aspects. It is to be understood that although preferred embodiments, specific constructions and configurations, as well as materials, have been discussed herein for devices according to the present invention, various changes or modifications in form and detail may be made without departing from the scope of this invention. For example, any formulas given above are merely representative of procedures that may be used. Functionality may be added or deleted from the block diagrams and operations may be interchanged among functional blocks. Steps may be added or deleted to methods described within the scope of the present invention.

Claims

1. A semiconductor cell culture device comprising: a. a semiconductor mesh (112) having islands (116) being interconnected by bridges (118) and defining through-pores (114) between the islands (116), wherein the mesh (112) has a top surface and a bottom surface, b. a cover (200) facing the top surface of the semiconductor mesh (112) and separated therefrom, thereby defining a space (202) for a cell culture (204), wherein the cover (200) has a top surface and a bottom surface, c. at least one mesh electrode (120) attached to the top surface or the bottom surface of the mesh, d. at least one cover electrode (220) attached to the bottom surface of the cover (200), e. first electrical connections (110) to one or more of the at least one mesh electrode (120), and f. second electrical connections (111) to one or more of the at least one cover electrode (220), wherein the first and second electrical connections (111) allow the performance of an electrical measurement between each of said one or more of the at least one mesh electrode (120) with a cover electrode (220), and wherein said one or more of the at least one mesh electrode (120) satisfy a geometrical condition imposing that at least one electrically conductive point (121) of each of said one or more of the at least one mesh electrode (120) must be located within a distance (d) of 100 pm from a through-pore (114).

2. The semiconductor cell culture device according to claim 1, wherein said distance (d) is 10 pm, preferably 5 pm from a through-pore (114).

3. The semiconductor cell culture device according to any one of the preceding claims, wherein said geometrical condition further imposes that at least 50%, preferably at least 75%, more preferably at least 90%, such as 100% of the top periphery (302) of said through-pore (114), said top periphery (302) being coplanar with that surface of the at least one mesh electrode (120) which is attached to the mesh, must be within the said distance (d) of at least one electrically conductive point of the at least one mesh electrode (120).

4. The semiconductor cell culture device according to any one of the preceding claims, wherein the geometrical condition further imposes that said through-pore (114) must have a top periphery (302) being coplanar with that surface of the at least one mesh electrode (120) which is attached to the mesh, said top periphery (302) having an average diameter such that the ratio between said average diameter (D) and the distance (d) separating said at least one electrically conductive point (121) from the through-pore is at least 2, preferably at least 3, more preferably at least 5.

5. The semiconductor cell culture device according to any one of the preceding claims, wherein the first and second electrical connections (111) allow the simultaneous performance of an electrical measurement between a same cover electrode (220) and a group of at least two of said one or more of the at least one mesh electrode (120), each electrode of said group satisfying said geometrical condition with respect to a same through- pore (114).

6. The semiconductor cell culture device according to claim 5, wherein said group is a group of all of said one or more of the at least one mesh electrode (120) satisfying said geometrical condition with respect to said same through-pore (114).

7. The semiconductor cell culture device according to any one of the preceding claims, wherein each through-pore has a top periphery (302), and wherein one or more of the at least one cover electrode (220) is positioned directly above: a. a through-pore such that a vertical projection of the electrode on the plane of the top surface of the mesh at least partially overlap with the through-pore, or b. a plurality of through-pores such that a vertical projection of the electrode on the plane of the top surface of the mesh entirely comprises the top periphery (302) of the plurality of through-pores.

8. The semiconductor cell culture device according to claim 7, wherein each of the at least one cover electrode (220) is positioned directly above a plurality of through- pores such that a vertical projection of the electrode on the plane of the top surface of the mesh entirely comprises the top periphery (302) of the plurality of through-pores and does not comprise at least some space between two adjacent through-pores.

9. The semiconductor cell culture device according to any one of the preceding claims, wherein the one or more of the at least one cover electrode (220) have a same shape and wherein the through-pores have said same shape.

10. The semiconductor cell culture device according to any one of the preceding claims, further comprising an electrical measurement system (117) connected to the first and second electrical connections (111), configured to perform measurements between any of the at least one mesh electrode (120) and a cover electrode (220).

11. The semiconductor cell culture device according to any one of the preceding claims wherein the electrical measurement is selected from a transepithelia I- transendothelial electrical measurement and an impedance spectroscopy measurement.

12. A method for detecting the presence of a fault in a biological barrier in a semiconductor cell culture device according to any one of the preceding claims, the method comprising: a. Initiating an electrical measurement sequentially between each of the one or more of the at least one mesh electrode (120) and a cover electrode (220) of the semiconductor cell culture device, thereby generating electrical outputs, b. Determining from the electrical outputs obtained from the electrical measurements which, if any of the one or more of the at least one mesh electrode (120) measure an output outside a predetermined value range, indicative of the presence of a fault in the cell culture (204) above a through-pore (114) within said distance (d) of these electrodes.

13. A semiconductor cell culture system comprising a device 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 to cause the semiconductor cell culture system of claim 13 to execute the steps of the method of claim 12.

15. A computer-readable medium having stored thereon the computer program of claim 14.