Single cell capture microfluidic calorimetry chip and preparation method thereof, and single cell capture microfluidic calorimetry system

Abstract

This invention provides a single-cell capture microchannel calorimetric chip, its fabrication method, and a single-cell capture microchannel calorimetric system, belonging to the field of semiconductor device technology. The single-cell capture microchannel calorimetric chip includes: a silicon substrate having opposing first and second surfaces, with two parallel-spaced receiving grooves disposed inside the silicon substrate between the first and second surfaces; and a protective film layer deposited on the first surface, the second surface, and the inner wall of the receiving grooves, the protective film layer within the receiving grooves surrounding and forming a microchannel unit; wherein, one end of the microchannel unit is provided with a microchannel inlet penetrating the second surface, the other end of the microchannel unit is provided with a microchannel outlet penetrating the second surface, and the portion of the microchannel unit located between the microchannel inlet and the microchannel outlet is provided with a cell capture chamber suitable for capturing cells.

Description

Technical Field

[0001] This invention belongs to the field of semiconductor device technology, specifically relating to a single-cell capture microfluidic calorimetric chip, its preparation method, and a single-cell capture microfluidic calorimetric system. Background Technology

[0002] All biological activities release or consume energy, which is used to maintain homeostasis and ultimately promote growth and development. Energy balance within an organism plays a crucial role in maintaining homeostasis, and monitoring this energy balance can provide information about cellular metabolism and biomolecular interactions. Cells primarily maintain energy balance through metabolic heat production; information on cellular metabolic heat can elucidate some fundamental cellular processes and reflect cellular activity.

[0003] However, monitoring heat changes during energy balance is challenging because some microorganisms produce or consume extremely small amounts of energy. Many methods have been developed to monitor energy balance and measure metabolic heat or temperature changes in organisms. Heat measurement from living organisms can be performed directly or indirectly, both of which involve various thermosensing techniques. Indirect measurement techniques mainly include thermometry based on luminescent materials and many other cell temperature sensing methods. Thermometry based on luminescent materials utilizes the fluorescence properties of luminescent materials to measure temperature changes in biological samples; however, this method is limited by the temperature resolution of the luminescent material and is easily affected by factors such as molecular concentration and cell composition. Direct measurement methods include contact thermometry using nanoscale thermal probes and microcalorimetry. Contact thermometry using nanoscale thermal probes typically measures intracellular temperature changes using thermocouples; however, during measurement, the thermal probe can damage the cell membrane, causing a cellular stress response and generating additional heat. Microcalorimetry is generally used to monitor the energy balance of organisms and measure heat changes within them. Among these methods, microcalorimetry has become a powerful and indispensable tool for characterizing biological metabolic processes in a label-free, non-invasive, and non-irritating manner. Microcalorimetry provides direct measurements of cellular heat information, thus offering a wealth of crucial information to reveal the underlying mechanisms of cellular metabolism. Microcalorimetry has been widely used for monitoring energy balance and measuring metabolic heat in biological processes within cells and organisms.

[0004] Buried channel technology is used to fabricate microstructures for fluid applications, such as silicon-based microchannels and connectors. This technology constructs microstructures by etching trenches, depositing capping layers on the trench sidewalls, removing the capping layer from the trench bottom, and etching the main body of the silicon substrate. The microstructures can be sealed by depositing appropriate capping layers to seal the trenches. Buried channel technology is a process that can be used to fabricate complete microchannels on a single wafer, requiring only a photomask and processing on one side of the wafer, without the need for assembly and bonding. This process results in a nearly flat and smooth substrate surface, facilitating further processing. Microchannels fabricated using buried channel technology have good sealing properties, but the etching rate is relatively slow when etching large-diameter, near-circular microchannels. Furthermore, it is difficult to operate when etching interconnected microchannels of different diameters, requiring multiple repetitions of the process. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a single-cell capture microfluidic calorimetric chip, its fabrication method, and a single-cell capture microfluidic calorimetric system, aiming to at least partially solve the above-mentioned technical problems. The specific technical solutions provided by this invention are as follows.

[0006] As a first aspect of the present invention, a single-cell capture microchannel calorimetric chip is provided, comprising: a silicon substrate having a first surface and a second surface opposite to each other, and two receiving grooves disposed parallel to each other inside the silicon substrate between the first surface and the second surface; and a protective film layer deposited on the first surface, the second surface and the inner wall of the receiving grooves, the protective film layer located in the receiving grooves surrounding and forming a microchannel unit; wherein, one end of the microchannel unit is provided with a microchannel inlet penetrating through the second surface, the other end of the microchannel unit is provided with a microchannel outlet penetrating through the second surface, and the portion of the microchannel unit located between the microchannel inlet and the microchannel outlet is provided with a cell capture chamber suitable for capturing cells.

[0007] As a second aspect of the present invention, a method for fabricating a single-cell capture microchannel calorimetric chip is provided, comprising: providing a silicon substrate with protective films deposited on opposing first and second surfaces; forming a first window on the protective film on the second surface of the silicon substrate, forming a slit structure on the protective film on the first surface of the silicon substrate, etching the interior of the silicon substrate through the first window and the slit structure to form two identical receiving grooves; depositing the same material as the protective film on the inner wall of the receiving grooves of the silicon substrate to form microchannel units, and sealing the slit structure; forming a temperature-sensing resistor and a heating resistor on the protective film on the first surface of the silicon substrate where the microchannel units are formed; forming a second window on the protective film on the first surface of the silicon substrate where the temperature-sensing resistor and the heating resistor are formed, and forming a heat-insulating groove surrounding the middle part of the microchannel unit inside the silicon substrate through the second window, thereby obtaining a single-cell capture microchannel calorimetric chip.

[0008] As a third aspect of the present invention, a single-cell capture microfluidic calorimetric system is provided, comprising: a single-cell capture microfluidic calorimetric chip, suitable for single-cell capture and single-cell metabolic heat measurement; a fluid drive device connected to the microfluidic inlet and microfluidic outlet of the single-cell capture microfluidic calorimetric chip, suitable for injecting cell suspension into the single-cell capture microfluidic calorimetric chip; and a data acquisition device electrically connected to the single-cell capture microfluidic calorimetric chip, suitable for receiving electrical signals transmitted by the single-cell capture microfluidic calorimetric chip.

[0009] Based on the above technical solutions, the single-cell capture microchannel calorimetric chip and its preparation method and the single-cell capture microchannel calorimetric system provided by the present invention have at least one of the following beneficial effects.

[0010] (1) In this embodiment of the invention, two identical receiving grooves are arranged in parallel inside a silicon substrate. Protective film layers are deposited on the opposing first and second surfaces of the silicon substrate and on the inner walls of the receiving grooves. The protective film layers deposited on the inner walls of the receiving grooves form microchannel units. The microchannel units are isolated from the external environment to avoid the influence of the external environment on cell metabolism during the measurement of cell metabolic heat. At the same time, the two microchannel units form a differential structure, which reduces common-mode interference during the measurement of cell metabolic heat and improves the measurement accuracy.

[0011] (2) In an embodiment of the present invention, a protective film layer is formed on a first surface and a second surface opposite to the silicon substrate to protect the silicon substrate during etching. A first window is formed on the protective film layer on the second surface, and a slit structure is formed on the protective film layer on the first surface. The interior of the silicon substrate is etched through the first window and the slit structure to form a receiving groove. Then, the same material as the protective film layer is deposited on the inner wall of the receiving groove to form a microfluidic unit. Etching the silicon substrate through the slit structure and the first window can reduce the chip fabrication process and improve the structural strength of the microfluidic unit. By adjusting the shape and size of the first window and the slit structure to control the shape and size of the receiving groove, the shape and size of the microfluidic unit can be adjusted to accommodate the capture of biological cells of different sizes. The first window and the slit structure are formed on the protective film layer on different surfaces to avoid the collapse of the microfluidic unit channel wall caused by the force applied by the external device when injecting or flowing out of the single-cell capture microfluidic calorimetric chip through the external device.

[0012] (3) In the embodiments of the present invention, the single-cell capture microfluidic calorimetric system provided by the present invention can realize the capture of cells in cell suspension and the accurate measurement of the metabolic heat of captured cells. It has the advantages of high sensitivity, small sample volume, fast response time and wide application. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of the overall structure of the single-cell capture microfluidic calorimetric chip in an embodiment of the present invention;

[0014] Figure 2 This is a detailed schematic diagram of the cell capture chamber of the microchannel unit in an embodiment of the present invention;

[0015] Figure 3 This is a detailed schematic diagram of the temperature measuring resistor and the heating resistor in an embodiment of the present invention;

[0016] Figure 4 This is a design diagram of a single-cell capture microfluidic calorimetric chip in an embodiment of the present invention;

[0017] Figure 5 This is a flowchart illustrating the fabrication process of the single-cell capture microfluidic calorimetric chip in an embodiment of the present invention.

[0018] Figure 6 This is a detailed design diagram of the temperature measuring resistor and the heating resistor in an embodiment of the present invention;

[0019] Figure 7 This is a schematic diagram of the structure of a microfluidic fixture holding a single-cell capture microchannel calorimetric chip in an embodiment of the present invention.

[0020] Explanation of reference numerals in the attached figures:

[0021] 100. Single-cell capture microfluidic calorimetric chip;

[0022] 1. Silicon substrate; 11. Microfluidic unit; 111. Cell capture chamber; 112. First microfluidic channel; 113. Second microfluidic channel; 12. Microfluidic channel inlet; 13. Microfluidic channel outlet; 14. Insulation groove; 15. Temperature sensing resistor; 16. Heating resistor;

[0023] 2. Protective film layer on the first surface;

[0024] 3. Protective film layer on the second surface. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0026] Existing single-cell capture microfluidic calorimetric chips mostly employ methods such as constructing microchannels on a silicon substrate or creating open microchannels on a silicon substrate and covering them with a glass substrate. The fabrication process for these chips is relatively cumbersome and complex, and the measurement of cellular metabolic heat after single-cell capture is inevitably affected by the external environment. Therefore, this application provides a single-cell capture microfluidic calorimetric chip, its fabrication method, and a single-cell capture microfluidic calorimetric system. Specifically, the single-cell capture microfluidic calorimetric chip of this application forms microchannel units within a silicon substrate, enabling single-cell capture and precise measurement of cellular metabolic heat.

[0027] Figure 1 This is a schematic diagram of the overall structure of the single-cell capture microfluidic calorimetric chip in an embodiment of the present invention.

[0028] As a first aspect of the present invention, a single-cell capture microfluidic calorimetric chip 100 is provided, such as Figure 1 As shown, it includes: a silicon substrate 1 having a first surface and a second surface opposite to each other, with two receiving grooves disposed parallel to each other inside the silicon substrate located between the first surface and the second surface; and a protective film layer deposited on the first surface, the second surface and the inner wall of the receiving grooves, the protective film layer located in the receiving grooves surrounding and forming a microchannel unit 11; wherein, one end of the microchannel unit is provided with a microchannel inlet 12 penetrating through the second surface, the other end of the microchannel unit is provided with a microchannel outlet 13 penetrating through the second surface, and the portion of the microchannel unit located between the microchannel inlet and the microchannel outlet is provided with a cell trapping chamber 111 suitable for trapping cells.

[0029] In this embodiment of the invention, two identical receiving grooves are arranged in parallel inside a silicon substrate. Protective films are deposited on the opposing first and second surfaces of the silicon substrate and on the inner walls of the receiving grooves. The protective films deposited on the inner walls of the receiving grooves form microchannel units. The microchannel units are isolated from the external environment to avoid the influence of the external environment on cell metabolism during the measurement of cellular metabolic heat. At the same time, the two microchannel units form a differential structure, reducing common-mode interference during the measurement of cellular metabolic heat and improving measurement accuracy.

[0030] Figure 2 This is a detailed schematic diagram of the cell capture chamber of the microchannel unit in an embodiment of the present invention.

[0031] According to embodiments of the present invention, such as Figure 2 As shown, the microchannel unit 11 includes a first microchannel 112 located between the microchannel inlet 12 and the cell capture chamber 111, and at least two second microchannels 113 located between the cell capture chamber 111 and the microchannel outlet 13; wherein the diameter of the first microchannel 112 is larger than the diameter of the cell, and the diameter of the second microchannel 113 is smaller than the diameter of the cell.

[0032] In an embodiment of the present invention, the cell suspension enters the microfluidic unit 11 through the microfluidic inlet 12. Since the diameter of the first microfluidic channel 112 is larger than the cell diameter, and the diameter of the second microfluidic channel 113 is smaller than the cell diameter, when the cell suspension flows from the first microfluidic channel 112 to the second microfluidic channel 113, the cells are captured by the cell capture chamber 111, and the remaining solution flows out through the second microfluidic channel 113, preventing blockage of the microfluidic unit 11. The microfluidic channel diameter can be adjusted according to the diameter of the cells in the introduced cell suspension; the present invention is not limited thereto.

[0033] According to embodiments of the present invention, such as Figure 2 As shown, the aforementioned single-cell capture microfluidic calorimetric chip further includes a heat insulation groove 14, disposed outside the cell capture cavity 111 inside the silicon substrate 1, suitable for isolating the cell capture cavity 111 from the silicon substrate 1. Because the heat insulation groove 14 isolates the cell capture cavity 111 from the silicon substrate 1, when measuring the metabolic heat of cells captured by the cell capture cavity 111, the influence of external temperature transmitted by the silicon substrate 1 on the cells can be further avoided, improving the accuracy of the measurement. The size and shape of the heat insulation groove 14 are not limited here; it only needs to not penetrate the silicon substrate 1 while isolating the cell capture cavity 111 from the silicon substrate 1.

[0034] Figure 3 This is a detailed schematic diagram of the temperature measuring resistor and the heating resistor in an embodiment of the present invention.

[0035] According to embodiments of the present invention, such as Figure 3 As shown, the aforementioned single-cell capture microfluidic calorimetric chip further includes a temperature-sensing resistor 15, located on the protective film layer 2 of the first surface, suitable for measuring the metabolic heat of cells and transmitting electrical signals to the outside. The temperature-sensing resistor 15 can be made of titanium or platinum, but the invention is not limited to these materials. The temperature-sensing resistor 15 is located on the cell capture chamber 111, and can measure the metabolic heat of cells captured in the cell capture chamber and transmit electrical signals to the outside through circuitry.

[0036] According to embodiments of the present invention, such as Figure 3 As shown, the aforementioned single-cell capture microfluidic calorimetric chip further includes a heating resistor 16, located on the protective film layer 2 of the first surface, suitable for providing stable temperature conditions for cell survival. The heating resistor 16 can be made of titanium or platinum, but the invention is not limited thereto. The heating resistor 16 is located on the cell capture chamber 111, opposite to the temperature measuring resistor, and heats the cell capture chamber 111 to provide suitable and stable temperature conditions for the cells within the cell capture chamber 111.

[0037] In embodiments of the present invention, the temperature-sensing resistor 15 and the heating resistor 16 can be configured in a serpentine structure to increase the coverage area, facilitating temperature measurement by the temperature-sensing resistor 15 and heating by the heating resistor 16. The materials of the temperature-sensing resistor 15 and the heating resistor 16 can be the same or different. When titanium (with a thermal conductivity of 15 W / (m·K) at 25°C and a temperature coefficient of resistance of 0.32% / K) is selected for both the temperature-sensing resistor 15 and the heating resistor 16, the line width of the temperature-sensing resistor 15 is set to be less than half the line width of the heating resistor 16. For example... Figure 3 As shown, conductive material is etched and deposited on the protective film layer of the first surface to achieve electrical connection between the temperature sensing resistor 15 and the heating resistor 16 and the external circuit. When the temperature sensing resistor 15 and the heating resistor 16 are made of titanium, a layer of titanium can be deposited before depositing the conductive material to improve the adhesion between the conductive material and the protective film layer and the conductivity with the titanium.

[0038] According to embodiments of the present invention, the protective film is made of either silicon nitride or silicon oxide. By depositing a protective film of low thermal conductivity silicon nitride or silicon oxide on both opposite surfaces of a silicon substrate, and simultaneously forming microfluidic units within the silicon substrate, the thermal conductivity of the single-cell capture microfluidic calorimeter chip can be further reduced, thus minimizing measurement errors.

[0039] Figure 4 This is a design diagram of a single-cell capture microfluidic calorimetric chip in an embodiment of the present invention; Figure 5 This is a flowchart illustrating the fabrication process of the single-cell capture microfluidic calorimetric chip in an embodiment of the present invention. Figure 6 This is a detailed design diagram of the temperature measuring resistor and the heating resistor in an embodiment of the present invention.

[0040] As a second aspect of the invention, according to... Figure 4 The structure shown is processed to provide a method for fabricating the above-mentioned single-cell capture microfluidic calorimetric chip, such as... Figure 5 As shown, it includes: a silicon substrate 1 with protective films deposited on its opposing first and second surfaces, such as... Figure 5 As shown in Figure a; a first window is formed on the protective film layer 3 on the second surface of the silicon substrate, and a slit structure is formed on the protective film layer 2 on the first surface of the silicon substrate. The interior of the silicon substrate is etched through the first window and the slit structure to form two identical receiving grooves, as shown in Figure a. Figure 5 b-5e; The same material as the protective film is deposited on the inner wall of the receiving trench on the silicon substrate to form microchannel units 11 and seal the gap structure, such as Figure 5 f; A temperature sensing resistor 15 and a heating resistor 16 are formed on the protective film layer 2 on the first surface of the silicon substrate 1 where the microfluidic unit 11 is formed, such as Figure 5g; A second opening is formed on the protective film layer 2 of the first surface of the silicon substrate 1, on which the temperature sensing resistor 15 and the heating resistor 16 are formed. Through the second opening, a heat insulation groove 14 is formed inside the silicon substrate 1, surrounding the middle part of the microchannel unit 11. Figure 5 h-5i yielded a single-cell capture microfluidic calorimetric chip 100.

[0041] In embodiments of the present invention, protective films are formed on a first surface and a second surface opposite to each other on a silicon substrate 1 to protect the silicon substrate 1 during etching. A first window is formed on the protective film 3 on the second surface, and a slit structure is formed on the protective film 2 on the first surface. The interior of the silicon substrate 1 is etched through the first window and the slit structure to form a receiving trench. Then, the same material as the protective film is deposited on the inner wall of the receiving trench to form a microfluidic unit 11. Etching the silicon substrate 1 through the slit structure and the first window can reduce the chip fabrication process and improve the structural strength of the microfluidic unit 11. By adjusting the shape and size of the first window and the slit structure to control the shape and size of the receiving trench, the shape and size of the microfluidic unit 11 can be adjusted to accommodate the capture of biological cells of different sizes. The first window and the slit structure are formed on protective films on different surfaces to avoid the collapse of the microfluidic unit channel wall caused by the force applied by the external device when injecting or flowing out a solution containing cells into or out of the single-cell capture microfluidic calorimetric chip.

[0042] In embodiments of the present invention, low-pressure chemical vapor deposition can be used to deposit the protective film layer, but the present invention is not limited thereto. Laser direct writing can be used to form the first window, the slit structure, the second window, the temperature sensing resistor, and the heating resistor, but the present invention is not limited thereto. Before etching the silicon substrate with the deposited protective film layer, a layer of metallic aluminum can be deposited on the surface of the protective film layer to protect it. Dry isotropic etching can be used to etch the receiving trenches for forming the microchannel inlet and outlet, and dry anisotropic etching can be used to etch the receiving trenches for forming the first and second microchannels and the heat insulation trench, but the present invention is not limited thereto. The first window is circular in shape to etch the microchannel inlet and outlet as circular as possible; the second window is rectangular in shape, but can also be adjusted according to the shape of the microchannel, but the present invention is not limited thereto.

[0043] According to an embodiment of the present invention, when forming the first microchannel 112, the slit structure is a structure with 3-7 slits interspersed; when forming the second microchannel 113, the slit structure is a structure with 1-3 slits interspersed. The internal width of the microchannel unit is controlled by adjusting parameters such as the slit path, number, and etching time of the slit structure to customize the microchannel diameter of a specific size.

[0044] For example, when the diameter of the cells in the cell suspension is 90-120 μm, a 4-inch double-polished silicon wafer with a 100-phase crystal structure and a diameter of 120-200 μm is used as the silicon substrate. The diameter of the first microchannel is set to be 120-200 μm, the diameter of the second microchannel is 20-90 μm, and the diameters of the microchannel inlet and outlet are the same, both 200-600 μm. Figure 6 As shown, when etching the receiving groove of the first microchannel, the slit structure consists of five slits intermittently distributed. When etching the receiving groove of the second microchannel, the slit structure consists of one slit intermittently distributed. The slit is 1 μm wide and 95 μm long. In the direction parallel to the etching path, the two slits are spaced 5 μm apart, and in the direction perpendicular to the etching path, the two slits are spaced 10 μm apart.

[0045] As a third aspect of the present invention, a single-cell capture microfluidic calorimetric system is provided, comprising: a single-cell capture microfluidic calorimetric chip 100, suitable for single-cell capture and single-cell metabolic heat measurement; a fluid drive device connected to the microfluidic inlet 12 and microfluidic outlet 13 of the single-cell capture microfluidic calorimetric chip 100, suitable for injecting cell suspension into the single-cell capture microfluidic calorimetric chip 100; and a data acquisition device electrically connected to the single-cell capture microfluidic calorimetric chip 100, suitable for receiving electrical signals transmitted by the single-cell capture microfluidic calorimetric chip 100.

[0046] In the embodiments of the present invention, the single-cell capture microfluidic calorimetric system provided by the present invention can realize the capture of cells in cell suspension and the accurate measurement of the metabolic heat of captured cells. It has the advantages of high sensitivity, small sample volume, fast response time and wide application.

[0047] Figure 7 This is a schematic diagram of the structure of a microfluidic fixture holding a single-cell capture microchannel calorimetric chip in an embodiment of the present invention.

[0048] According to an embodiment of the present invention, the single-cell capture microchannel calorimetry system further includes: a microfluidic clamp for fixing the single-cell capture microchannel calorimetry chip, thereby enabling communication between the single-cell capture microchannel calorimetry chip and the fluid drive device. Figure 7 As shown, the microfluidic fixture holds the single-cell capture microchannel calorimetric chip within two structures, with rubber plugs at the microchannel inlet and outlet to connect them to the fluid drive device.

[0049] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A single-cell capture microfluidic calorimetric chip, comprising: A silicon substrate having opposing first and second surfaces, with two receiving grooves disposed parallel to each other inside the silicon substrate between the first and second surfaces; A protective film layer is deposited on the first surface, the second surface, and the inner wall of the receiving tank. The protective film layer located in the receiving tank and the protective film layer located on the first surface surround and form a microchannel unit. as well as The temperature-sensing resistor, located on the protective film layer of the first surface, is suitable for measuring the metabolic heat of cells and transmitting electrical signals to the outside world; The microchannel unit has a microchannel inlet penetrating the second surface at one end and a microchannel outlet penetrating the second surface at the other end. The portion of the microchannel unit located between the microchannel inlet and the microchannel outlet has a cell trapping chamber suitable for trapping cells. The protective film is made of either silicon nitride or silicon oxide.

2. The chip according to claim 1, wherein, The microchannel unit includes a first microchannel located between the microchannel inlet and the cell capture chamber, and at least two second microchannels located between the cell capture chamber and the microchannel outlet; The diameter of the first microchannel is larger than the diameter of the cell, and the diameter of the second microchannel is smaller than the diameter of the cell.

3. The chip according to claim 1, further comprising: A heat insulation groove is disposed inside the silicon substrate on the outside of the cell capture cavity, which is suitable for isolating the cell capture cavity from the silicon substrate.

4. The chip according to claim 1, further comprising: The heating resistor, located on the protective film layer of the first surface, is suitable for providing stable temperature conditions for the cell to survive.

5. A method for fabricating a chip as described in claim 1, comprising: A silicon substrate with protective films deposited on its opposing first and second surfaces is provided; A first window is formed on the protective film layer on the second surface of the silicon substrate, and a slit structure is formed on the protective film layer on the first surface of the silicon substrate. The interior of the silicon substrate is etched through the first window and the slit structure to form two identical receiving grooves. The same material as the protective film layer is deposited on the inner wall of the receiving groove in the silicon substrate to form microchannel units and seal the gap structure; A temperature-sensing resistor is formed on a protective film layer on the first surface of the silicon substrate in which microchannel units are formed, to obtain a single-cell capture microchannel calorimetric chip.

6. The preparation method according to claim 5 further includes: A second opening is formed on the protective film layer of the first surface of the silicon substrate on which the temperature-sensing resistor is formed, and a heat insulation groove is formed inside the silicon substrate through the second opening, surrounding the middle part of the microchannel unit.

7. The preparation method according to claim 5 further includes: A heating resistor is formed on a protective film layer on the first surface of the silicon substrate in which microchannel units are formed.

8. The preparation method according to claim 5, wherein, The microchannel unit includes a first microchannel located between the microchannel inlet and the cell capture chamber, and at least two second microchannels located between the cell capture chamber and the microchannel outlet; When the first microchannel is formed, the slit structure is a structure in which 3-7 slits are distributed alternately; When forming the second microchannel, the slit structure is a structure in which 1-3 slits are distributed alternately.

9. A single-cell capture microfluidic calorimetric system, comprising: The chip as described in any one of claims 1-4 is suitable for single-cell capture and single-cell metabolic heat measurement; A fluid drive device, connected to the microchannel inlet and microchannel outlet of the chip, is suitable for injecting cell suspension into the chip; as well as A data acquisition device, electrically connected to the chip, is suitable for receiving electrical signals transmitted by the chip.

10. The system according to claim 9, further comprising: A microfluidic fixture is used to fix the chip in order to enable communication between the chip and the fluid drive device.

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