KR20220031532A - Cell behavior analysis module, method for manufacturing cell behavior analysis module, and cell behavior analysis system comprising the same - Google Patents

Abstract

The present invention discloses: a substrate; an insulating layer disposed on the substrate; a plurality of nanostructures disposed on the substrate to have a pixel array pattern and exposed over the insulating layer; a cell support film, disposed on the insulating layer and the nanostructure, which supports a cell; a first electrode connected to the substrate; and a second electrode connected to the nanostructure. The substrate is composed of a p-type semiconductor. The nanostructure is composed of an n-type semiconductor. Disclosed are a cell behavior analysis module, a method for manufacturing the cell behavior analysis module, and a cell behavior analysis system comprising the same.

Description

Cell behavior analysis module, method for manufacturing cell behavior analysis module, and cell behavior analysis system comprising the same

The present invention relates to a cell behavior analysis module, a method for manufacturing a cell behavior analysis module, and a cell behavior analysis system comprising the same.

Cell behavior is one of the key factors that must be studied to understand the intracellular environment and intracellular mechanisms. Cell behavior is not only an important process in forming and maintaining the human body, but is also related to disease states such as cancer cell behavior and cancer metastasis, so it has been extensively studied in the fields of new drugs, disease treatment, and tissue engineering.

There are many ways to control the behavior of cells by fabricating surfaces using biocompatible polymers (biopolymers) and changing the shape and chemical properties of the surface using chemical coatings, photolithography, micro-contact printing, and micro-molding, etc. Studies are underway. Since many mammalian cells function after attachment to the surface, the interaction between the cell and the surface is a very important factor in the field of biotechnology.

8Since the surface affects cellular behaviors such as survival, differentiation, cytoplasmic division, polarization and behavior, changing the properties of the surface can affect the behavior of cells.

However, these prior studies focus on controlling the behavior of cells, but research on the technology to analyze and monitor the behavior of cells is insufficient.

An object of the present invention is to provide a cell behavior analysis module capable of precisely analyzing cell behavior by increasing the degree of integration of a light emitting device, a method for manufacturing a cell behavior analysis module, and a cell behavior analysis system including the same.

On the other hand, the technical problems to be achieved in the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned are clearly to those of ordinary skill in the art to which the present invention belongs from the description below. can be understood

A cell behavior analysis module according to an embodiment of the present invention includes a substrate; an insulating layer disposed on the substrate; a plurality of nanostructures disposed on the substrate to have a pixel array pattern and exposed over the insulating layer; a cell support film disposed on the insulating layer and the nanostructure and supporting cells; a first electrode connected to the substrate; and a second electrode connected to the nanostructure, wherein the substrate may be formed of a p-type semiconductor, and the nanostructure may be formed of an n-type semiconductor.

In addition, each of the nanostructures may have a width of 500 nm to 1000 nm, and an interval between the plurality of nanostructures may be 2 μm to 5 μm.

In addition, it may further include a housing disposed on the cell support film and providing a space in which the living environment of the cell is simulated.

In addition, a plurality of microlenses disposed on the cell support film to correspond to the respective positions of the nanostructures may be further included.

On the other hand, the manufacturing method of the cell behavior analysis module according to an embodiment of the present invention comprises the steps of disposing a seed layer and a first electrode of an n-type semiconductor on a substrate of the p-type semiconductor; disposing a mask on the substrate to cover the seed layer; forming a plurality of holes having a pixel array pattern in the mask; growing the n-type semiconductor seed layer through the plurality of holes to form a nanostructure of an n-type semiconductor; disposing an insulating layer on the substrate such that an upper end of the nanostructure is exposed; disposing a second electrode on the insulating layer to be electrically connected to the nanostructure; and disposing a cell support film on the insulating layer and the nanostructure.

In addition, in the step of forming the nanostructures, each of the nanostructures may have a width of 500nm to 1000nm, and an interval between the plurality of nanostructures may be 2μm to 5μm.

In addition, in the step of disposing the cell support film, the cell support film may include a plurality of microlenses disposed on the cell support film to correspond to the respective positions of the nanostructures.

On the other hand, the cell behavior analysis system according to an embodiment of the present invention includes a cell behavior analysis module; an illuminance measuring unit measuring each pixel and illuminance of each of the nanostructures to generate illuminance information for each pixel; and a controller configured to receive and store the illuminance information from the illuminance measurement unit, and analyze the illuminance change for each pixel according to time to analyze cell behavior.

Also, when the illuminance change value of a specific pixel is equal to or greater than a preset threshold, the controller may determine that the nanostructure of the corresponding pixel is defective.

In addition, the control unit calculates a total amount of illuminance values of all pixels at a first time point when cells are not arranged, calculates a total amount of illuminance values of all pixels at a second time point at which cells are arranged, the first time point and the first time point The size of the cells may be calculated based on the difference in the total amount of illuminance between the two time points.

In addition, the control unit calculates a total amount of illuminance values of all pixels at a third time point after the second time point, and calculates the growth degree of the cells based on a difference between the total amount of illuminance values between the second time point and the third time point can do.

According to an embodiment of the present invention, it is possible to precisely analyze the behavior of cells by increasing the degree of integration of the light emitting device.

On the other hand, the effects obtainable in the present invention are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those of ordinary skill in the art to which the present invention belongs from the description below. will be able

1 is an exemplary view showing a cell behavior analysis system according to an embodiment of the present invention,
2 and 3 are exemplary views for explaining a method of manufacturing a cell behavior analysis device in the cell behavior analysis system according to an embodiment of the present invention;
4 and 5 are SEM images showing a cell behavior analysis device in the cell behavior analysis system according to an embodiment of the present invention,
6 is an SEM image for explaining the light emission state of the cell behavior analysis device in the cell behavior analysis system according to an embodiment of the present invention;
7 is a graph showing the luminosity of a cell behavior analysis device in a cell behavior analysis system according to an embodiment of the present invention;
8 is an exemplary view showing a cell behavior analysis system according to another embodiment of the present invention,
9 is a front view (left) and a side view (right) showing the structure of a cell support film and microlens in the cell behavior analysis system of FIG. 8 .

Hereinafter, an embodiment of the present invention will be described in more detail with reference to the accompanying drawings. Embodiments of the present invention may be modified in various forms, and the scope of the present invention should not be construed as being limited to the following embodiments. This embodiment is provided to more completely explain the present invention to those of ordinary skill in the art. Accordingly, the shapes of elements in the drawings are exaggerated to emphasize a clearer description.

The configuration of the invention for clarifying the solution to the problem to be solved by the present invention will be described in detail with reference to the accompanying drawings based on a preferred embodiment of the present invention, but the same in assigning reference numbers to the components of the drawings For the components, even if they are on different drawings, the same reference numbers are given, and it is noted in advance that the components of other drawings can be cited when necessary in the description of the drawings.

1 is an exemplary diagram illustrating a cell behavior analysis system according to an embodiment of the present invention, and FIGS. 2 and 3 illustrate a method of manufacturing a cell behavior analysis device in a cell behavior analysis system according to an embodiment of the present invention 4 and 5 are SEM images showing a cell behavior analysis device in a cell behavior analysis system according to an embodiment of the present invention, and FIG. 6 is a cell behavior analysis system according to an embodiment of the present invention. It is an SEM image for explaining the light emission state of the cell behavior analysis device, and FIG. 7 is a graph showing the luminosity of the cell behavior analysis device in the cell behavior analysis system according to an embodiment of the present invention.

First, referring to FIG. 1 , a cell behavior analysis system according to an embodiment of the present invention may include a cell behavior analysis module 100 , an illuminance measurement unit 200 , and a control unit 300 .

The cell behavior analysis module 100 may include a cell behavior analysis device 110 , a cell support film 120 , and a housing 130 .

Meanwhile, referring to FIGS. 2 to 7 , the cell behavior analysis device 110 includes a substrate 10 , a first electrode 20 , a nanostructure 30 , an insulating layer 50 , and a second electrode 60 . may include

On the other hand, the cell behavior analysis device 110 according to the present invention can be composed of a device that emits light with a band gap difference when electrons flow by bonding a semiconductor having a band gap of gallium nitride (GaN) and zinc oxide (ZnO) series. there is.

Gallium nitride (GaN) and zinc oxide (ZnO)-based materials have a large energy bandgap of direct transition type, and have recently been researched and developed as a light source in the ultraviolet (UV), blue (Blue), and green (Green) wavelength ranges. This is being done and is being commercialized.

However, in the present invention, the constituent materials of the cell behavior analysis device 110 are not limited to gallium nitride (GaN) and zinc oxide (ZnO), and may be formed of various combinations of semiconductor materials.

The substrate 10 is preferably made of p-type gallium nitride (GaN), and more preferably a p-GaN layer is formed on an Al ₂ O ₃ substrate or a sapphire substrate.

The first electrode 20 is an electrode for connecting the substrate 10 to an external power source, and may include, for example, a conductive material such as Pd, Au, Ni, Al, Pt, or Cr.

The nanostructure 31 may be disposed on the substrate 10 in a nano-rod shape, and may be formed of n-type zinc oxide (ZnO).

The nanostructure 31 may have a pixel array pattern. That is, the plurality of nanostructures 31 may be spaced apart from each other in a lattice form. Meanwhile, although not shown, the plurality of nanostructures 31 may be disposed to be spaced apart from each other in a radial pattern with the center of the substrate 10 as a center.

Here, it is preferable that each of the nanostructures 31 has a width of 500 nm to 1000 nm, and an interval between the adjacent nanostructures 31 is 2 μm to 5 μm.

Through this, it is possible to indicate a change in the amount of light by controlling the size of the pixel, and since the inter-pixel interference does not occur at an interval of 1 μm or more between the structures, the resolution can be adjusted according to the interval.

The insulating layer 50 is disposed on the substrate 10 in the form of enclosing the plurality of nanostructures 30 , and is made of an acrylic resin-based material such as polymethyl methacrylate (PMMA), polyethylene terephthalate (PET), poly carbonate (PC), and COC. (cyclic-olefin copolymer) and may include one of polyethylene naphthalate (PEN) resin.

Meanwhile, the insulating layer 50 is an impermeable (Photoresist, etc.) or semi-transmissive material (PMMA, SU-8, SiO ₂ , etc.) It is preferably composed of

The second electrode 60 is an electrode for connecting the nanostructure 31 to an external power source, is disposed on the insulating layer 50, and is a layered common electrode connecting the upper ends of the plurality of nanostructures 31 Preferably, it is composed of a transparent electrode and can emit light propagating from the nanostructure 31 to the outside.

Meanwhile, it is preferable that light propagating from the top of the nanostructure 31 is not diffused, and the second electrode 60 is preferably made of a material having a small refractive index difference from that of the nanostructure 31 .

The cell support film 120 may be disposed on the second electrode 60 to support the cell 1 . That is, the cell 1 may be seated on the cell support film 120 and move. Here, the cell support film 120 is composed of a biocompatible polymer thin film, and preferably has light transmittance.

The housing 130 is disposed on the cell support film 120 , and may provide a space in which the living environment of the target cell 1 is simulated. Here, the housing 130 may contain a culture medium, and the housing 130 may be connected to a separate biological environment simulating device (not shown) to continuously receive an environment suitable for the life of the cells 1 . .

Referring back to FIG. 1 , the illuminance measuring unit 200 may be disposed on the cell behavior analysis module 100 to measure the illuminance of light emitted from the cell behavior analysis device 110 .

The illuminance measuring unit 200 may be implemented as an illuminance sensor or an image sensor, but the present invention is not specifically limited thereto.

The illuminance measuring unit 200 may recognize each of the plurality of nanostructures 31 as a pixel in the cell behavior analysis device 110 , and may define coordinates of the plurality of pixels.

Also, the illuminance measuring unit 200 may classify illuminance for each pixel and generate illuminance information for each pixel. The illuminance information for each pixel generated by the illuminance measuring unit 200 may be transmitted to the controller 300 through a wired or wireless method.

The control unit 300 may be implemented as a program mounted on a MICOM or may be implemented as a chip-on-board type through each chipset on a substrate.

The controller 300 may analyze the behavior of the cell 1 based on the illuminance information for each pixel received from the illuminance measuring unit 200 .

Specifically, the controller 300 may quantify the illuminance information for each pixel, and may analyze the behavior of the cell 1 based on the illuminance change over time.

For example, the control unit 300 stores the illuminance for each pixel at the first time point when no cells are disposed and sets it as a default value. Thereafter, when cells are not arranged for several seconds (1 second to 60 seconds) at the first time point, the range of the default value may be corrected by measuring the change in illuminance for each pixel. In this case, the default value is set as a range, which can improve reliability in the analysis of cell behavior compared to being limited to a specific value.

Thereafter, the control unit 300 calculates the change in the illuminance value for each pixel compared to the default value at the second time point at which the cells are arranged, so as to specify the pixel in which the illuminance value has changed, size can be specified.

Here, the control unit 300 may quantify the cell permeability ratio through the changed illuminance value at the second time point. For example, the controller 300 may quantify the transmittance ratio of the cell by dividing the illuminance value measured at the pixel whose illuminance value is changed at the second time point by the median value of the default range at the first time point. In this case, the controller 300 may recognize that the pixel in which the illuminance value is reduced by the cell permeability ratio is located in the cell.

Through this, the controller 300 may define a pixel in which the illuminance value is reduced by the cell permeability ratio, calculate the change in the illuminance value of each pixel, and analyze the cell behavior (movement and size change).

In addition, the control unit 300 may set the calculated transmittance ratio of the cells as a threshold value, and when the change in the illuminance value at a specific time point or a specific pixel exceeds the threshold value, it is determined that an error has occurred in the corresponding pixel and the corresponding pixel The illuminance value of can be excluded from the measurement.

In addition, the controller 300 calculates the total amount of illuminance values of all pixels at a first time point when cells are not disposed, calculates the total amount of illuminance values of all pixels at a second time point at which cells are disposed, the first time point and the second time point The size of the cells may be calculated based on the difference in the total amount of illuminance between time points.

Specifically, the control unit 300 may calculate the cell size through the following [Equation 1].

[Equation 1]

S ₁ = D ² XN

N = (AB) / (L ₁ -L ₂ )

Here, S ₁ is the cell size (area) at the second time point, D is the length of the pixel plane (=distance between nanostructures), N is the number of pixels with reduced illuminance values, A is the total amount of illuminance values at the first time point, B is the total amount of illuminance values at the second time point, L ₁ is the basic illuminance value of the unit pixel at the first time point, and L ₂ is the illuminance value of a specific unit pixel decreased by the transmittance ratio at the second time point.

In addition, the control unit may calculate the total amount of illuminance values of all pixels at a third time point after the second time point, and calculate the growth degree of the cells based on a difference value of the total amount of illuminance values between the second time point and the third time point. there is.

Specifically, the control unit 300 may calculate the growth degree of the cells through the following [Equation 2].

[Equation 2]

S ₂ -S ₁ = S _C

S ₂ = D ² XN ₂

N ₂ = (BC) / (L ₁ -L ₃ )

Here, S ₁ - is the cell size (area) at the second time point, S ₂ is the cell size (area) at the third time point, S _C is the cell growth size (area), and C is the illuminance value at the third time point The total amount, L ₁ represents the basic illuminance value of the unit pixel at the first time point, and L ₃ represents the illuminance value of the specific unit pixel decreased by the transmittance ratio at the third time point.

In addition, the controller 300 may calculate the growth rate of cells grown for a specific period by dividing the growth degree of cells calculated through [Equation 2] by time.

Hereinafter, a method of manufacturing a cell behavior analysis device will be described in detail with reference to FIGS. 2 to 7 .

First, a GaN (p-type semiconductor substrate) 10 is deposited on an Al ₂ O ₃ substrate by CVD. After that, a 50 nm thick Au layer with a 10 nm Ni adhesive layer is deposited using an evaporator to form a grid electrode (first electrode) 20 .

Thereafter, a ZnO seed layer (n-type seed layer) 30 is formed through spin coating.

Thereafter, after removing the photoresist, a positive electron beam resist mask (poly methyl methacrylate (PMMA)) 40 is coated to form a nano hole array 41 having a diameter of 500 nm and a periodic rate of 5 μm by electron beam lithography. The nano hole array 41 has a pixel array pattern.

To form ZnO of n-tpye on a masked GaN substrate, put it in a solution containing 40 mM hexamethylene tetramine (HMTA) (Sigma-Aldrich) and 40 mM zinc nitride (Sigma-Aldrich), and oven at 85 °C for 4 An array of nanowires (n-type nanostructures) 31 was grown for a period of time. The c-axis GaN positions exposed to the aqueous solution grow into epitaxial ZnO nanowires, thus producing an array of single ZnO nanowires uniformly patterned in centimeters.

Then, an insulating poly(methyl methacrylate) (PMMA) layer (insulating layer) 50 is spin-coated to wrap the grown ZnO nanowires.

Thereafter, since the tip of the ZnO nanowire should be exposed according to the device structure, the upper part of the PMMA is etched by an oxygen plasma process.

Finally, an ITO film with a thickness of about 200 nm is deposited to form an upper common electrode (second electrode) 60 by magnetron sputtering to achieve a heterostructure device.

8 is an exemplary view illustrating a cell behavior analysis system according to another embodiment of the present invention, and FIG. 9 is a front view (left) and It is a side view (right).

8 and 9 , the cell behavior analysis system according to another embodiment of the present invention has a different configuration of the microlens 410 than the cell behavior analysis system according to the embodiment of the present invention shown in FIG. 1 . Therefore, in the following, only the configuration of the differentiated micro lens 410 will be described in detail, and detailed descriptions of reference numerals overlapping the same configuration will be omitted.

The microlens 410 is formed on the inner or outer surface of the cell support film 120, and may be disposed to correspond to the position and size of the plurality of nanostructures.

That is, the plurality of micro lenses 410 may be disposed to have a pixel array pattern, and each of the plurality of micro lenses 410 may face each of the plurality of nano structures.

On the other hand, the plurality of micro lenses 410 may be converged by converging the light emitted from the nanostructure.

Here, the light converged through the plurality of microlenses 410 may be emitted in a smaller area, so that the light emitted from the adjacent nanostructures does not interfere with each other and may be clearly distinguished from each other. That is, since the illuminance measuring unit can more clearly distinguish pixels and reduce errors due to optical interference, more reliable analysis results can be derived.

The above detailed description is illustrative of the present invention. In addition, the above description shows and describes preferred embodiments of the present invention, and the present invention can be used in various other combinations, modifications, and environments. That is, changes or modifications are possible within the scope of the concept of the invention disclosed herein, the scope equivalent to the written disclosure, and/or within the scope of skill or knowledge in the art. The written embodiment describes the best state for implementing the technical idea of the present invention, and various changes required in specific application fields and uses of the present invention are possible. Accordingly, the detailed description of the present invention is not intended to limit the present invention to the disclosed embodiments. Also, the appended claims should be construed to include other embodiments.

100: Cell behavior analysis module
200: illuminance measuring unit
300: control unit

Claims (11)

Board;
an insulating layer disposed on the substrate;
a plurality of nanostructures disposed on the substrate to have a pixel array pattern and exposed over the insulating layer;
a cell support film disposed on the insulating layer and the nanostructure and supporting cells;
a first electrode connected to the substrate; and
and a second electrode connected to the nanostructure,
The substrate is composed of a p-type semiconductor, and the nanostructure is a cell behavior analysis module composed of an n-type semiconductor.
The method of claim 1,
Each of the nanostructures has a width of 500 nm to 1000 nm,
An interval between the plurality of nanostructures is a cell behavior analysis module of 2 μm to 5 μm.
The method of claim 1,
The cell behavior analysis module further comprising a housing disposed on the cell support film and providing a space in which the living environment of the cell is simulated.
The method of claim 1,
The cell behavior analysis module further comprising a plurality of microlenses disposed on the cell support film to correspond to the respective positions of the nanostructures.
disposing a seed layer and a first electrode of an n-type semiconductor on a substrate of a p-type semiconductor;
disposing a mask on the substrate to cover the seed layer;
forming a plurality of holes having a pixel array pattern in the mask;
growing the n-type semiconductor seed layer through the plurality of holes to form a nanostructure of an n-type semiconductor;
disposing an insulating layer on the substrate so that an upper end of the nanostructure is exposed;
disposing a second electrode on the insulating layer to be electrically connected to the nanostructure; and
A method of manufacturing a cell behavior analysis module comprising disposing a cell support film on the insulating layer and the nanostructure.
6. The method of claim 5,
In the step of forming the nanostructure,
Each of the nanostructures has a width of 500 nm to 1000 nm,
The spacing between the plurality of nanostructures is a method of manufacturing a cell behavior module of 2 μm to 5 μm.
6. The method of claim 5,
In the step of disposing the cell support film,
The cell support film,
A method of manufacturing a cell behavior analysis module comprising a plurality of microlenses disposed on the cell support film to correspond to the respective positions of the nanostructures.
The cell behavior analysis module according to any one of claims 1 to 4;
an illuminance measuring unit measuring each pixel and illuminance of the nanostructures to generate illuminance information for each pixel; and
The cell behavior analysis system further comprising a controller that receives and stores the illuminance information from the illuminance measurement unit and analyzes the illuminance change for each pixel over time to analyze the cell behavior.
9. The method of claim 8,
The control unit is a cell behavior analysis system for determining that the nanostructure of the pixel is defective when the illuminance change value of the specific pixel is greater than or equal to a preset threshold.
9. The method of claim 8,
The control unit is
Calculate the total amount of illuminance values of all pixels at the first time point in which cells are not arranged,
Calculate the total amount of illuminance values of all pixels at the second time point when the cells are arranged,
A cell behavior analysis system for calculating the size of the cell based on the difference in the total amount of illuminance values between the first time point and the second time point.
11. The method of claim 10,
The control unit is
calculating the total amount of illuminance values of all pixels at a third time point after the second time point;
A cell behavior analysis system for calculating the growth degree of the cells based on the difference in the total amount of illuminance values between the second time point and the third time point.

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