A cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging

Abstract

The present invention discloses a cell in-situ temperature-controlled chip culture system adapted for microscopic optical imaging, and relates to the technical field of in-situ observation of microscopic live imaging. The key points of the technical solution are: comprising a temperature-controlled chip and an accessory for cell culture, wherein the temperature-controlled chip is detachably connected to the accessory and is located inside the accessory; the accessory comprises a cell culture device, a connection card, a control circuit, a culture pool with openings at both ends, a connector and a connection interface; the temperature-controlled chip comprises a transparent chip body, a temperature-measuring layer and a heating layer, wherein the temperature-measuring layer and the heating layer are respectively fixedly connected to the opposite side walls of the chip body. The system is a device adapted for an optical microscope observation system, which can monitor and precisely control the temperature of the chip control area in real time, and is applied to various experiments and application scenarios that require precise control of cell culture, microbial incubation, material temperature control, and in-situ observation of cells, and provides powerful tools and support for in-depth research in the fields of life sciences and materials science.

Description

Technical Field

[0001] The present invention relates to the technical field of cell culture, and more specifically, to an in-situ temperature-controlled cell chip culture system adapted for microscopic optical imaging. Background Art

[0002] Temperature control plays a vital role in the study of physiological activities and metabolic processes of cells and microorganisms, as well as in the study of the physical and chemical properties of materials. Under different temperature conditions, the fluidity of cell membranes, the catalytic rate of enzymes, and material transfer will change. The simulation of environmental temperature can study the biological processes of cells and microorganisms adapting to and responding to temperature changes, explore the biological characteristics of cells and pathogens, the activity of drugs, and the response of the immune system. At the same time, in the field of materials science, temperature is an important factor and key parameter that affects the thermal conductivity and thermal expansion, crystal growth, and phase change behavior of materials. Temperature control can be used to explore the physical and chemical properties of materials, the structural evolution of crystals, the kinetics of reactions, and the thermodynamic properties. By controlling the temperature, changes in the internal environment and external stimuli of organisms can be simulated, as well as the behavior and properties of materials at specific temperatures can be explored, which helps to deeply understand the basic principles of life sciences and materials sciences, and provide a theoretical basis and experimental basis for related applications and innovations.

[0003] In the study of the life activities of cells and microorganisms, temperature directly affects the physiological activity of living organisms. In the biomedical field, microscopic imaging of living cell samples is often required. Currently, optical microscopy is an important technical observation method in biomedical and material science research. At the same time, temperature control is a key control requirement in the above scientific research and technological applications, involving multiple aspects such as imaging quality, cell physiology and sample stability.

[0004] The temperature control of common microscopes is mainly achieved through temperature control tables or thermostats. A constant temperature environment is set in the experimental equipment, and the sample temperature is controlled by circulating the gas in the thermostat. The temperature control tables and thermostats are large in size, and the temperature control accuracy of the gas circulation system is limited. It is impossible to achieve precise control of local temperature, and it is not convenient to observe the changes of samples under different temperature conditions in real time under a microscope. In the research of life sciences and materials sciences, temperature control and sample observation are very important experimental means. Traditional temperature control equipment has problems such as large size and complex operation, which is not conducive to in-situ observation under an optical microscope.

[0005] Therefore, based on the above problems, the applicant provides a temperature control chip system with compact structure, simple operation, and the ability to accurately control temperature and realize in-situ observation. Summary of the invention

[0006] The purpose of the present invention is to provide a cell in situ temperature-controlled chip culture system adapted to microscopic optical imaging, which can realize rapid and precise temperature control in in vitro experiments, and can also realize dynamic observation of samples during temperature control, so as to facilitate the exploration of the biological characteristics of cells and the physical and chemical properties of materials, and contribute to a deep understanding of the basic principles in the fields of life sciences and materials sciences, and provide a theoretical basis and experimental basis for related applications and innovations.

[0007] The above technical objectives of the present invention are achieved through the following technical solutions: a cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging, comprising a temperature-controlled chip and an accessory for cell culture, wherein the temperature-controlled chip is detachably connected to the accessory and is located inside the accessory;

[0008] The accessory comprises a cell culture device, a connection card, a control circuit, a culture tank with openings at both ends, a connector and a connection interface; the control circuit is fixedly connected to the cell culture device, the connection card is detachably connected to the cell culture device, the control circuit and the connection card are both located inside the cell culture device, the temperature control chip is fixedly connected to the connection card, the culture tank is fixedly connected to the temperature control chip, the input end of the connection interface is located on the outer side wall of the cell culture device, and the output end thereof is fixedly connected to the input end of the control circuit, and the two ends of the connector are respectively connected to the output end of the control circuit and the temperature control chip;

[0009] The temperature control chip comprises a transparent chip body, a temperature measuring layer and a heating layer. The temperature measuring layer and the heating layer are respectively fixedly connected to the opposite side walls of the chip body, and the temperature measuring layer is located on the side wall of the chip body close to the culture pool.

[0010] The present invention is further configured as follows: a reinforcement protection sheet is provided around the culture pool, and the bottom of the reinforcement protection sheet is fixedly connected to the temperature control chip.

[0011] The present invention is further configured as follows: the chip body is made of a transparent material with good optical imaging properties, such as glass with a thickness of 50-200 um.

[0012] The present invention is further configured such that: the culture pool is made of biocompatible material.

[0013] The present invention is further configured as follows: the temperature measuring layer and the heating layer are composed of temperature sensitive materials and are used for real-time monitoring and control of the chip temperature.

[0014] The present invention is further configured as follows: the control circuit adopts PID temperature control technology and PWM heating technology to perform temperature control and heating.

[0015] In summary, the present invention has the following beneficial effects:

[0016] 1. The culture system can perform precise temperature control. By adopting advanced micro-nano processing technology, dense heating areas and temperature measurement areas are integrated on the temperature control chip, ensuring the stability of sample temperature in the culture area of ​​the temperature control chip, providing a reliable basic platform for precise temperature control research in cell experiments;

[0017] 2. The culture system has convenient optical observation effects. The ultra-thin glass substrate design of the temperature control chip realizes the direct optical observation requirements of the temperature control area under the microscope. The temperature control chip can adapt to the optical requirements of different imaging platforms such as optical microscopes and ultra-high resolution microscopes. At the same time, the glass substrate of the temperature control chip is evenly etched, and the design reduces optical distortion during imaging, providing higher-resolution images, which is conducive to the study of dynamic observation of cells, microorganisms, and materials science;

[0018] 3. The culture system is highly adaptable and integrated. The size of the temperature control chip is the same as that of a universal cover glass. The related accessories of the culture system provide fixed support for the culture and incubation of samples. It can be flexibly adapted to the optical microscope stage on the market and support all aspects of cell biology and materials research.

[0019] 4. Compared with traditional incubators, this cell culture system is compact, adaptable to the microscope stage, and easy to operate, which enables users to heat conveniently in a limited space, improving the user experience and work efficiency;

[0020] 5. The culture system has a precise and controllable temperature change process. The temperature control chip of the system adopts a PID temperature control system, which can realize a programmable temperature change process. This design can realize programmed temperature control, simulate different speeds and durations of temperature rise and fall, and meet the temperature change and temperature control needs in different fields such as life sciences and materials science;

[0021] 6. The culture system has a fast temperature change system. The temperature control chip adopts PWM temperature heating technology. The temperature change per second can exceed 100°C, which can achieve extremely fast temperature rise, which is very beneficial for time-sensitive experiments and research;

[0022] 7. The culture system has brought beneficial effects in terms of accuracy of temperature control, convenience of optical observation, adaptability of the system, compact size and ease of operation, controllable temperature change process and the ability to quickly reach the target temperature. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is a schematic diagram of the appearance of a chip culture system in an embodiment of the present invention;

[0024] Figure 2 is a 3D schematic diagram of a temperature control chip in an embodiment of the present invention;

[0025] Figure 3 is a top view of a chip culture system in an embodiment of the present invention;

[0026] Figure 4 is an axonometric diagram of a chip culture system according to an embodiment of the present invention;

[0027] Figure 5 It is a linear graph of the test data of the unloaded chip in the embodiment of the present invention.

[0028] In the figure: 1. Temperature control chip; 2. Cell culture device; 3. Connection card; 4. Connection interface; 5. Culture pool; 6. Connector. DETAILED DESCRIPTION

[0029] The following is combined with Figure 1-5 The present invention is described in further detail.

[0030] Embodiment: A cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging, such as Figures 1 to 5 As shown, it includes a temperature control chip 1 and accessories for cell culture. The temperature control chip 1 is manufactured using micro-nano processing technology. The chip size is 12mm×12mm, and the size of the heating area is 5mm×5mm. The chip undergoes strict quality control to ensure that its quality is optimal. It provides a reliable basic platform for cell culture and observation.

[0031] The temperature control chip 1 is used in combination with accessories, and the accessories specifically include a cell culture device 2 that can be adapted to a microscope, a connecting card 3, a control circuit, a culture pool 5 with openings at both ends, a connector 6 and a connecting interface 4, wherein the control circuit is fixedly connected to the cell culture device 2, the connecting card 3 is detachably connected to the cell culture device 2, the control circuit and the connecting card 3 are both located inside the cell culture device 2, and the temperature control chip 1 is fixedly connected to the connecting card 3; the culture pool 5 is made of biocompatible material, and PDMS (polydimethylsiloxane) material is preferably used in this embodiment, and its bottom is fixedly connected to the upper surface of the temperature control chip 1, and the input end of the connecting interface 4 is located on the outer wall of the cell culture device 2, and its output end is fixedly connected to the input end of the control circuit.

[0032] The accessories provide fixed support for the temperature control chip 1 and retain gas exchange and liquid exchange openings, which can provide complete cell research tools and environment, support all aspects of cell biology research, and help scientific researchers conduct accurate and reliable cell experiments and research.

[0033] The temperature control chip 1 specifically includes a transparent chip body, a temperature measuring layer and a heating layer, wherein the chip body is made of a transparent material with good optical imaging properties. In this embodiment, a No. 1.5 glass with a thickness of 0.17 mm is preferred, which is consistent with the thickness of the confocal dish. This specific thickness is very critical for obtaining high-quality images, because the dish used in the confocal microscope usually has a standard bottom thickness of 0.17 mm, which is the same as the thickness of a standard microscope slide. Most microscope objective lenses are optimized for this thickness. If higher resolution is required, a thinner bottom will be used, which makes the temperature control chip 1 perfectly matched with various microscope equipment, so as to achieve The purpose of high-resolution microscope is to take cell images, so that the device is perfectly matched with various microscope equipment, realizing real-time high-resolution shooting under the microscope. Users can obtain clear and accurate images and data when observing tiny structures or conducting delicate experiments, providing strong support for scientific research and application practice; in addition, the application of ultra-thin glass makes the temperature control chip 1 light and portable, easier to carry and install, further expanding its scope of application; this feature combined with excellent heating performance and comprehensive real-time monitoring functions makes the temperature control chip 1 an ideal choice in various industries, bringing users a convenient and efficient use experience and meeting diverse needs.

[0034] The temperature measuring layer is composed of temperature measuring film lines, and the heating layer is composed of temperature measuring film lines. Both the temperature measuring film lines and the temperature measuring film lines are composed of temperature sensitive materials, and in this embodiment, the temperature sensitive material is preferably platinum metal material; the temperature measuring layer and the heating layer are fixedly connected to the opposite side walls of the chip body respectively, for realizing real-time monitoring and control of the chip temperature, and the temperature measuring layer is located on the side wall of the chip body close to the culture pool 5; wherein the temperature control chip 1 is manufactured using photolithography technology, and a series of processes such as design and plate making, film preparation, photoresist coating, photolithography exposure, development and etching, metallization, removal of residual photoresist, testing and packaging can accurately manufacture a complex temperature control chip 1, so that it is possible to create a complex heating circuit in a very small space, and realize efficient and precise thermal control to ensure that the cells are heated evenly during use.

[0035] Preferably, in this embodiment, a reinforcing protective sheet made of acrylic or PC material is fixedly installed around the culture pool 5, and a through groove is arranged in the middle of the reinforcing protective sheet. The size of the through groove is the same as the size of the outer wall of the culture pool 5, and the inner wall and bottom of the through groove of the reinforcing protective sheet are respectively bonded to the outer wall of the culture pool 5 and the upper surface of the temperature control chip 1, so as to better fix the culture pool 5.

[0036] This embodiment is preferred. In terms of temperature control, our control circuit uses RTD temperature measurement technology and PWM heating technology to achieve rapid adjustment and precise control of temperature. RTD temperature measurement technology uses the characteristics of metal resistance temperature sensors to provide high-precision temperature measurement; the temperature control chip 1 can sense the working temperature in real time and transmit it to the control circuit through signals. The control circuit makes timely adjustments through the embedded logic algorithm; at the same time, through PWM heating technology, the temperature control chip 1 can quickly heat up at a rate of 100℃ / s and quickly reach the set temperature, greatly improving the heating efficiency and saving time costs; through experimental testing of the unloaded chip, the unloaded chip test data linear graph obtained is as follows Figure 5 As shown, it reflects that the temperature control chip 1 culture system is capable of precise temperature control, ensuring the stability of the sample temperature in the culture area of ​​the temperature control chip 1, and providing a reliable basic platform for precise temperature control research in cell experiments.

[0037] Through precise temperature control technology, the temperature control chip 1 can maintain a stable heating state, ensuring that the temperature fluctuation during the heating process is extremely small, and the temperature control accuracy reaches 0.1°C. PWM heating uses transistors to quickly switch the on and off states of the current, thereby generating a series of current pulses. The duration (width) of these pulses can be precisely controlled, thereby precisely controlling the heat generated by the heater; PWM technology allows rapid adjustment of the pulse width and rapid response to temperature changes, ensuring the accuracy and rapid response of temperature control. The temperature control chip 1 can control the power switching time with a certain duty cycle, quickly adjust the heating power, and achieve precise control of the temperature to meet the needs of rapid temperature changes in different situations.

[0038] In addition, the control circuit also uses PID algorithm for remote temperature feedback adjustment. The PID algorithm monitors temperature changes in real time and combines the three control parameters of proportion, integration and differentiation to adjust the heating power in a timely and accurate manner, so that the temperature can quickly return to the set value and remain stable. Users can monitor and adjust the working status of the temperature control chip 1 in real time through the remote controller to achieve remote intelligent control; the application of this temperature control technology enables our temperature control chip 1 to achieve efficient and precise temperature control in various application scenarios, with a temperature control accuracy of 0.1°C and a temperature control accuracy of up to 0.2°C, providing a programmable, intelligent and convenient temperature control solution.

[0039] In terms of heating, the temperature control chip 1 also adopts a design that combines the Peltier effect and resistive heating to achieve efficient and uniform heating effects; the Peltier effect uses the thermoelectric effect generated when current passes through two materials with different thermal conductivity to achieve rapid cooling and heating; resistive heating uses the principle of generating heat by passing current through a conductive material to provide stable heating capabilities.

[0040] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. A cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging, characterized by: It comprises a temperature control chip (1) and an accessory for cell culture, wherein the temperature control chip (1) is detachably connected to the accessory and is located inside the accessory; The accessory comprises a cell culture device (2), a connection card (3), a control circuit, a culture tank (5) with openings at both ends, a connector (6) and a connection interface (4); the control circuit is fixedly connected to the cell culture device (2), the connection card (3) is detachably connected to the cell culture device (2), the control circuit and the connection card (3) are both located inside the cell culture device (2), the temperature control chip (1) is fixedly connected to the connection card (3), the culture tank (5) is fixedly connected to the temperature control chip (1), the input end of the connection interface (4) is located on the outer side wall of the cell culture device (2), and the output end thereof is fixedly connected to the input end of the control circuit, and the two ends of the connector (6) are respectively connected to the output end of the control circuit and the temperature control chip (1); The temperature control chip (1) comprises an optically transparent chip body, a temperature measuring layer and a heating layer. The temperature measuring layer and the heating layer are made of thin films of temperature-sensitive materials and are respectively fixedly connected to the opposite side walls of the chip body. The temperature measuring layer is located on the side wall of the chip body close to the culture pool (5).

2. The cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging according to claim 1, characterized in that: A reinforcement protection sheet is provided around the culture pool (5), and the bottom of the reinforcement protection sheet is fixedly connected to the temperature control chip (1).

3. The cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging according to claim 1, characterized in that: The chip body is made of glass with good optical imaging properties and a thickness of 50-200 um.

4. The cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging according to claim 1, characterized in that: The culture pool (5) is made of biocompatible materials.

5. The cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging according to claim 1, characterized in that: The temperature measuring layer and the heating layer are composed of temperature sensitive materials and are used for real-time monitoring and control of the chip temperature.

6. The cell in situ temperature-controlled chip culture system adapted for microscopic optical imaging according to claim 1, characterized in that: The control circuit adopts PID temperature control technology and PWM heating technology to perform temperature control and heating.

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