Long-time cell culture device based on micro-fluidic chip

Abstract

The utility model discloses a long-time cell culture device based on a micro-fluidic chip, which comprises a placement frame, a culture box is arranged on the top surface of the placement frame, and a sample box frame is arranged below the culture box; the space in the culture box is a sterile water bin, sterile water is contained in the sterile water bin, and the cell micro-fluidic chip is completely immersed in the sterile water. According to the utility model, the cell micro-fluidic chip is completely immersed in sterile water, so that the phenomenon that nano bubbles are formed in the micro-fluidic chip after long-time cell culture is effectively avoided, the influence of the bubbles caused by humidity change of an external environment on cell activity is eliminated, and the state of cells is ensured.

Description

Technical Field

[0001] The utility model belongs to the technical field of microfluidic chip cell culture, relates to a microfluidic chip cell culture device, and specifically relates to a long-term cell culture device based on a microfluidic chip. Background Art

[0002] Live cell detection can provide real-time data of cell responses. Combined with microfluidic technology, it can perform high-throughput drug screening and other applications on living cells. When conducting live cell experiments on microfluidic chips, it is necessary to maintain a constant temperature and humidity environment to keep the cells stable for a long time.

[0003] The existing technology has the following main defects: First, when using microfluidic chips for long-term cell culture, the liquid in the microfluidic chip will evaporate through the inherent nanopores in polydimethylsiloxane, forming nanobubbles in the chip channel, thereby affecting the cell state. Second, the existing microfluidic chip cell culture devices are mostly split structures with low integration, large space occupation, and inconvenient use. Summary of the invention

[0004] In view of the shortcomings of the prior art, the purpose of the utility model is to provide a long-term cell culture device based on a microfluidic chip, so as to solve the technical problem that when a microfluidic chip is used for long-term cell culture in the prior art, the liquid in the microfluidic chip evaporates to form nanobubbles, thereby affecting the cell state.

[0005] In order to solve the above technical problems, the utility model adopts the following technical solutions:

[0006] A long-term cell culture device based on a microfluidic chip comprises a placement rack, a culture box is arranged on the top surface of the placement rack, and a sample box rack is arranged below the culture box.

[0007] The culture box includes a conductive heat transfer bottom plate, a cell microfluidic chip is fixedly arranged in the middle position of the conductive heat transfer bottom plate, four transparent side panels are arranged on the conductive heat transfer bottom plate around the cell microfluidic chip along the vertical direction, the four transparent side panels are symmetrically arranged in pairs, and a transparent top panel is detachably arranged on the top of the four transparent side panels; four transparent side panels are arranged on the top surface of the conductive heat transfer bottom plate along the vertical direction; the space enclosed by the conductive heat transfer bottom plate, the four transparent side panels and the transparent top panel is a sterile water tank, sterile water is contained in the sterile water tank, and the cell microfluidic chip is completely immersed in the sterile water.

[0008] The sample box rack comprises a sample box rack bottom plate, on the top surface of which four sample box rack side plates are fixedly arranged in the vertical direction, the four sample box rack side plates are symmetrically arranged in pairs, and a sample box rack top plate is detachably arranged at the top of the four sample box rack side plates; a plurality of sample tube placement openings are respectively provided on the lateral sides of the sample box rack top plate, the sample tube placement openings are located on the lateral outer sides of the sample box rack side plates, sample tubes are arranged in the sample tube placement openings, and the liquid outlet end of the sample tube is connected to the liquid inlet end of the cell microfluidic chip.

[0009] A temperature control chip is arranged on the top plate of the sample box rack; a conductive metal sheet is arranged on each of the lateral sides of the conductive heat transfer bottom plate, a thermistor is fixedly arranged on the surface of the conductive heat transfer bottom plate, and the thermistor is close to the cell microfluidic chip; the signal input end of the temperature control chip is connected to the signal output end of the thermistor, and the signal output end of the temperature control chip is connected to the conductive metal sheet; the space enclosed by the sample box rack top plate, four sample box rack side plates and the sample box rack bottom plate is a battery compartment, and a battery is arranged in the battery compartment; the current output end of the battery is connected to the current input end of the temperature control chip.

[0010] The utility model also has the following technical features:

[0011] The sample box rack has a length of 9 cm, a width of 7 cm and a height of 2.7 cm; the culture box has a length of 11 cm, a width of 7 cm and a height of 2 cm; the placement rack has a length of 16 cm, a width of 12 cm and a height of 7 cm.

[0012] Two current output interfaces and one temperature detection interface are arranged on the side panel of the sample box rack located on the longitudinal front side; a first wire is arranged in the current output interface, and the temperature control chip is connected to the conductive metal sheet through the first wire; a second wire is arranged in the temperature detection interface, and the temperature control chip is connected to the thermistor through the second wire.

[0013] Temperature control chip switches are arranged on the longitudinal front and rear side panels of the sample box rack.

[0014] Charging interfaces are arranged on the longitudinal front and rear side panels of the sample box frame, and the current input end of the battery is connected to the charging interface.

[0015] A culture box window is respectively provided on both lateral sides of the transparent top plate, and a liquid infusion tube and a gas infusion tube are arranged in the culture box window; the sample tube is connected to the cell microfluidic chip through the liquid infusion tube, and the air inlet end of the sample tube is connected to the air outlet end of the gas infusion tube.

[0016] The sample tube comprises a sample tube body, which is closed around and at the bottom, and open at the top. A sample tube cover is detachably mounted on the open top of the sample tube body, and the sample tube body is used to contain cell fluid.

[0017] An infusion needle and a gas delivery needle are arranged on the sample tube cover. The bottom end of the infusion needle extends into the cell fluid, and the top end of the infusion needle passes through and extends out of the sample tube cover. The infusion needle located above the sample tube cover is connected to the cell microfluidic chip through the infusion tube; the bottom end of the gas delivery needle extends out of the sample tube body, and the top end of the gas delivery needle passes through and extends out of the sample tube cover. The gas delivery needle located above the sample tube cover is connected to the gas delivery tube.

[0018] The placement rack includes a placement rack top plate and a placement rack bottom plate, and multiple support legs are installed between the placement rack top plate and the placement rack bottom plate, and the support legs are arranged along the vertical direction; a culture box is arranged on the top surface of the placement rack top plate, and a sample box rack is arranged on the top surface of the placement rack bottom plate.

[0019] Compared with the prior art, the utility model has the following technical effects:

[0020] (I) The utility model completely immerses the cell microfluidic chip in sterile water, thereby avoiding the phenomenon of nanobubbles being formed in the microfluidic chip after long-term cell culture, eliminating the influence of bubbles on cell activity caused by changes in external environmental humidity, and ensuring the state of the cells.

[0021] (II) The utility model integrates the cell culture control part (temperature control chip, conductive metal sheet, thermistor and battery) and the sample tube into the sample box rack, which reduces the space occupied by the entire device and improves the convenience of use.

[0022] (III) The top plate and side plates of the culture box of the utility model are both transparent plates, and the bottom plate of the culture box is nearly transparent ITO glass. This transparent structure facilitates imaging using a microscope platform during the culture process; in addition, the transparent side plates of the culture box are conducive to the experimenter to observe the water level of the sterile water at any time and replenish the sterile water in time to ensure that the microfluidic chip is always completely immersed in the sterile water during the long-term culture process.

[0023] (IV) The device of the utility model uses a rechargeable battery to power the temperature control chip, and can achieve long-term temperature control even when the AC power is off or there is no AC power, thereby improving the convenience of use and environmental applicability of the device. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Schematic diagram of the overall structure of the long-term cell culture device based on microfluidic chips.

[0025] Figure 2Schematic diagram of the structure of the culture box.

[0026] Figure 3 Schematic diagram of the structure of the sample box rack.

[0027] Figure 4 Schematic diagram of the sample tube structure.

[0028] Figure 5 is the evaporation rate of PDMS microfluidic chip under different environmental conditions

[0029] The meanings of the numbers in the figure are: 1-placing rack, 2-culture box, 3-sample box rack, 4-cell microfluidic chip, 5-sample tube, 6-temperature control chip, 7-conductive metal sheet, 8-thermistor, 9-battery, 10-first wire, 11-second wire, 12-infusion tube, 13-gas tube.

[0030] 101-top plate of the placement rack, 102-bottom plate of the placement rack, 103-supporting legs.

[0031] 201 - conductive heat transfer bottom plate, 202 - transparent side plate, 203 - transparent top plate, 204 - culture box window.

[0032] 301 - sample box rack bottom plate, 302 - sample box rack side plate, 303 - sample box rack top plate, 304 - sample tube placement port, 305 - current output interface, 306 - temperature detection interface, 307 - temperature control chip switch, 308 - charging interface.

[0033] 501 - sample tube body, 502 - sample tube cover, 503 - infusion needle, 504 - gas infusion needle.

[0034] The specific contents of the utility model are further explained in detail below in conjunction with the embodiments. DETAILED DESCRIPTION

[0035] In the present invention, "long-term cell culture" refers to a cell culture time of ≥48h.

[0036] It should be noted that all components in the present invention, unless otherwise specified, are components known in the art, such as:

[0037] The conductive heat transfer bottom plate 201 adopts a conventional ITO (indium tin oxide) glass plate known in the prior art.

[0038] The cell microfluidic chip 4 adopts a conventional cell microfluidic chip known in the prior art, and the cell microfluidic chip is made by reverse molding of polydimethylsiloxane.

[0039] The temperature control chip 6 adopts a conventional temperature controller known in the prior art, and the temperature control chip is a W1209 digital display temperature controller module. The working voltage of the temperature control chip is 12V, the temperature measurement accuracy is 0.1°C, the control accuracy is 0.1°C, and the refresh frequency is 0.5S.

[0040] The thermistor 8 is a conventional NTC thermistor known in the prior art, with a resistance value of 10KΩ, a resistance accuracy of ±0.01, a temperature measurement range of -55°C to 125°C, and a resistance B value of 3950.

[0041] The following are specific embodiments of the present invention. It should be noted that the present invention is not limited to the following specific embodiments, and all equivalent changes made on the basis of the technical solution of the present application fall within the protection scope of the present invention.

[0042] Example:

[0043] This embodiment provides a long-term cell culture device based on a microfluidic chip, such as Figure 1 As shown, it comprises a placement rack 1 , a culture box 2 is arranged on the top surface of the placement rack 1 , and a sample box rack 3 is arranged below the culture box 2 .

[0044] like Figure 1 and Figure 2 As shown, the culture box 2 includes a conductive heat transfer bottom plate 201, a cell microfluidic chip 4 is fixedly arranged in the middle position of the conductive heat transfer bottom plate 201, four transparent side panels 202 are arranged along the vertical direction on the conductive heat transfer bottom plate 201 around the cell microfluidic chip 4, the four transparent side panels 202 are symmetrically arranged in pairs, and a transparent top panel 203 is detachably arranged on the top of the four transparent side panels 202; four transparent side panels 202 are arranged on the top surface of the conductive heat transfer bottom plate 201 along the vertical direction; the space enclosed by the conductive heat transfer bottom plate 201, the four transparent side panels 202 and the transparent top panel 203 is a sterile water tank, sterile water is contained in the sterile water tank, and the cell microfluidic chip 4 is completely immersed in the sterile water.

[0045] like Figure 1 and Figure 3As shown, the sample box rack 3 includes a sample box rack bottom plate 301, on the top surface of the sample box rack bottom plate 301, four sample box rack side plates 302 are fixedly arranged along the vertical direction, the four sample box rack side plates 302 are symmetrically arranged in pairs, and the top of the four sample box rack side plates 302 is detachably provided with a sample box rack top plate 303; the lateral sides of the sample box rack top plate 303 are respectively provided with a plurality of sample tube placement openings 304, the sample tube placement openings 304 are located on the lateral outer sides of the sample box rack side plates 302, and the sample tubes 5 are arranged in the sample tube placement openings 304, and the liquid outlet end of the sample tube 5 is connected to the liquid inlet end of the cell microfluidic chip 4; the sample box rack top plate 303 A temperature control chip 6 is arranged on it; a conductive metal sheet 7 is arranged on each of the lateral sides of the conductive heat transfer bottom plate 201, and a thermistor 8 is fixedly arranged on the surface of the conductive heat transfer bottom plate 201, and the thermistor 8 is close to the cell microfluidic chip 4; the signal input end of the temperature control chip 6 is connected to the signal output end of the thermistor 8, and the signal output end of the temperature control chip 6 is connected to the conductive metal sheet 7; the space enclosed by the sample box rack top plate 303, the four sample box rack side plates 302 and the sample box rack bottom plate 301 is a battery compartment, and a battery 9 is arranged in the battery compartment; the current output end of the battery 9 is connected to the current input end of the temperature control chip 6.

[0046] In this embodiment, the size of the conductive heat transfer bottom plate 201 is equal to that of a 96-well plate, and the size is: length 128 mm×width 85 mm×thickness 1 mm, and the resistance value is: 7 -10 The transparent side plate 202 and the transparent top plate 203 are made of transparent acrylic plates with a thickness of 2 mm. Waterproof silicone is used for sealing protection between the conductive heat transfer bottom plate 201. The cell microfluidic chip 4 is fixed on the conductive heat transfer bottom plate 201 by plasma bonding. After the cell microfluidic chip 4 is fixed on the conductive heat transfer bottom plate 201, the conductive heat transfer bottom plate 201 is used to provide a culture temperature for the cell microfluidic chip 4.

[0047] In this embodiment, the sample box rack top plate 303, the sample box rack side plates 302 and the sample box rack bottom plate 301 are made of polylactic acid material or photosensitive resin.

[0048] In this embodiment, the number of the sample tube placement openings 304 is 10, and the number of the sample tube placement openings 304 can be set according to actual needs.

[0049] In this embodiment, the cell microfluidic chip 4 has a size of 7 cm in length × 5 cm in width × 1 cm in height, and includes several cell culture chambers with a length, width and height of 200 um × 200 um × 50 um. The cell culture chambers are connected by channels with a width and height of 100 um × 25 um, and samples are input and output through the inlet and outlet holes.

[0050] In this embodiment, the temperature control chip 6 is embedded in the top plate 303 of the sample box rack. The temperature control chip 6 includes a temperature display, a function selection button, and a temperature setting plus or minus button. The temperature control chip 6 can display the real-time temperature of the microfluidic chip and control the on and off of the current on the conductive heat transfer bottom plate 201.

[0051] In this embodiment, the battery 9 is embedded in the sample box frame bottom plate 301 and can be recharged and reused. Its power supply voltage is 4 to 12V and is used to provide a 12V working voltage to the temperature control chip 6 .

[0052] As a specific solution of this embodiment, the dimensions of the sample box rack 3 are: length 9 cm × width 7 cm × height 2.7 cm; the dimensions of the culture box 2 are: length 11 cm × width 7 cm × height 2 cm, and this dimension design can ensure that there is enough space to hold water and the cell microfluidic chip 4; the dimensions of the placement rack 1 are: length 16 cm, width 12 cm, height 7 cm.

[0053] As a specific solution of this embodiment, Figure 1 and Figure 3 As shown, two current output interfaces 305 and one temperature detection interface 306 are provided on the sample box rack side plate 302 located on the longitudinal front side; a first wire 10 is provided in the current output interface 305, and the temperature control chip 6 is connected to the conductive metal sheet 7 through the first wire 10; a second wire 11 is provided in the temperature detection interface 306, and the temperature control chip 6 is connected to the thermistor 8 through the second wire 11.

[0054] In this embodiment, the thermistor 8 is installed on the surface of the conductive heat transfer bottom plate 201, connected to the second wire 11 extending from the temperature detection interface 306, and connected to the temperature detection electrode of the temperature control chip 6 in the sample box rack 3. The temperature control chip 6 is set to 37°C, and the measured temperature is displayed in real time on the display screen of the temperature control chip 6. The output positive and negative electrodes of the temperature control chip 6 are connected to the conductive metal sheets 7 at both ends of the conductive heat transfer bottom plate 201 through the first wire 10 via the current output interface. When the displayed temperature is lower than the set temperature of the temperature control chip 6, the two sides of the conductive heat transfer bottom plate 201 are powered on. When the displayed temperature is higher than the set temperature of the temperature control chip 6, the power is turned off, thereby maintaining a stable temperature environment for the cell microfluidic chip 4 when culturing cells.

[0055] As a specific solution of this embodiment, Figure 3 As shown, a temperature control chip switch 307 is provided on the sample box rack side panels 302 located at the front and rear in the longitudinal direction. The temperature control chip switch 307 is used to control the opening and closing of the temperature control chip 6.

[0056] As a specific solution of this embodiment, Figure 3As shown, a charging interface 308 is provided on the sample box rack side panels 302 located at the front and rear in the longitudinal direction, and the current input end of the battery 9 is connected to the charging interface 308 . When the battery 9 is low on power, the battery 9 is charged through the charging interface 308 .

[0057] As a specific solution of this embodiment, Figure 2 and Figure 4 As shown, a culture box window 204 is respectively provided on both lateral sides of the transparent top plate 203, and a liquid infusion tube 12 and a gas infusion tube 13 are provided in the culture box window 204; the sample tube 5 is connected to the cell microfluidic chip 4 through the liquid infusion tube 12, and the air inlet end of the sample tube 5 is connected to the air outlet end of the gas infusion tube 13.

[0058] In this embodiment, the culture box window 204 is 3 cm long and 0.5 cm wide, and is disposed 1 cm away from the edge of the transparent top plate 203. The liquid infusion tube 12 and the gas infusion tube 13 are transparent rubber tubes with an outer diameter of 1.65 mm and an inner diameter of 0.55 mm.

[0059] In this embodiment, the sample tube 5 is filled with a cell suspension or a fresh cell culture medium. After ventilation is introduced into the sample tube 5 through the gas supply tube 13, the cell suspension or the fresh cell culture medium flows from the infusion tube 12 into the cell microfluidic chip 4 under the action of gas pressure, thereby achieving cell inoculation or fluid replacement.

[0060] As a specific solution of this embodiment, Figure 2 and Figure 4 As shown, the sample tube 5 includes a sample tube body 501, which is closed on all sides and at the bottom, and has an open top. A sample tube cover 502 is detachably mounted on the open top of the sample tube body 501, and the sample tube body 501 is used to hold cell fluid; an infusion needle 503 and a gas supply needle 504 are arranged on the sample tube cover 502, the bottom end of the infusion needle 503 extends into the cell fluid, the top end of the infusion needle 503 passes through and extends out of the sample tube cover 502, and the infusion needle 503 located above the sample tube cover 502 is connected to the cell microfluidic chip 4 through the infusion tube 12; the bottom end of the gas supply needle 504 extends out of the sample tube body 501, the top end of the gas supply needle 504 passes through and extends out of the sample tube cover 502, and the gas supply needle 504 located above the sample tube cover 502 is connected to the gas supply tube 13.

[0061] In this embodiment, the middle part of the sample tube cover 502 is made of rubber material, which is convenient for the infusion needle 503 and the gas delivery needle 504 to pass through. The infusion needle 503 and the gas delivery needle 504 are made of stainless steel, with an outer diameter of 0.7 mm and an inner diameter of 0.4 mm. The infusion tube 12 is tightly wrapped outside the infusion needle 503. The infusion needle 503 and the infusion tube 12 are used to transport the cell fluid in the sample tube 5 into the cell microfluidic chip 4. The infusion needle 503 is embedded in the inlet and outlet holes of the cell microfluidic chip 4. The gas delivery tube 13 is tightly wrapped outside the gas delivery needle 504. The gas delivery needle 504 and the gas delivery tube 13 are used to input a mixed gas into the sample tube 5. The mixed gas is a gas mixed with air and carbon dioxide at a ratio of 19:1, and the air pressure introduced into the sample tube 5 is 1psi.

[0062] As a specific solution of this embodiment, as shown in Figure 1, the placement rack 1 includes a placement rack top plate 101 and a placement rack bottom plate 102, and multiple support legs 103 are installed between the placement rack top plate 101 and the placement rack bottom plate 102, and the support legs 103 are arranged along the vertical direction; a culture box 2 is arranged on the top surface of the placement rack top plate 101, and a sample box rack 3 is arranged on the top surface of the placement rack bottom plate 102. In this embodiment, the culture box 2 and the sample box rack 3 are placed in the upper and lower layers of the placement rack 1, which can reduce the overall occupied space of the device.

[0063] The assembly and working process of the utility model are as follows:

[0064] First, cover the edge of the conductive heat transfer bottom plate 201 with a circle of 8mm wide transparent tape; spin-coat a PDMS thin layer on the conductive heat transfer bottom plate 201 at a rotation speed of 1500rpm, and after removing the transparent tape, expose the edge of the conductive heat transfer bottom plate 201; use plasma etching (total processing time 60s) to stick the cell microfluidic chip 4 on the spin-coated PDMS thin layer; paste the transparent side plate 202 on the ITO glass to form a water tank, and finally install the transparent top plate 203 to complete the assembly and production of the culture box.

[0065] Second, inject sterilized deionized water into the water tank through the culture box window 204 reserved on the top cover, and stop when the water tank is full without bubbles, to ensure that the cell microfluidic chip 4 is completely immersed in the liquid; connect one end of the infusion tube 12 and the air tube 13 to the inlet and outlet holes of the cell microfluidic chip 4, and then connect the other end of the infusion tube 12 and the air tube 13 to the 1.25mL sample tube 5.

[0066] Third, turn on the temperature control chip switch 307, adjust the temperature control chip 6, adjust the temperature to 37°C, and preheat for a period of time (the preheating time depends on the ambient temperature); after the temperature stabilizes at 37°C, apply positive pressure (1psi) to the sample tube 5, and the pressure squeezes the cell suspension in the sample tube 5 into the infusion tube 12. The cell suspension is injected into the temperature control chip 6 from the infusion needle 503 through the infusion tube 12. After the channel and cell culture chamber in the cell microfluidic chip 4 are filled, maintain the pressure in the sample tube 5, open the liquid inlet of the cell microfluidic chip 4 for 1 hour, and ensure that the channel of the cell microfluidic chip 4 is filled with culture medium without bubbles.

[0067] Fourth, perform cell culture: observe the temperature display to ensure that it is stable at 37° C. Replace the sample tube 5 connected to the cell microfluidic chip 4 with fresh culture medium, and replace the culture medium every hour under pressure. The cell culture process can be photographed under an optical microscope.

[0068] The utility model carries out the following experimental design to prove the effect of the above device:

[0069] Experimental Group 1: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, and the temperature of the incubator was controlled at 37°C; the humidity was maintained at a moderate indoor condition: ~30% RH relative humidity; an external air line was attached to the surface of the chip and blown from one long side to the other side, and the pressure gauge controlled the flow rate to: ~0.05m / s.

[0070] Experimental group 2: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art. The temperature of the incubator was controlled at 37° C. and the humidity was maintained at moderate indoor conditions: ˜30% RH; no airflow.

[0071] Experimental Group 3: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, and the temperature of the incubator was controlled at 37°C and the humidity was maintained at a moderate indoor condition: ~30% RH; before the experiment, the cell microfluidic chip 4 was submerged in water for 24 hours so that the micro-nano pores in the PDMS material were filled with water molecules; there was no airflow.

[0072] Experimental group 4: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art. The temperature of the incubator was controlled at 37° C. and the humidity was maintained at a moderate indoor condition: 30% RH; the microfluidic chip was immersed below the oil layer.

[0073] Experimental Group 5: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, the temperature of the incubator was controlled at 37°C, and the humidity was maintained at a moderate indoor condition: ~30% RH relative humidity; the microfluidic chip was immersed below the oil layer; before the experiment, the cell microfluidic chip 4 was submerged in water for 24 hours so that the micro-nano pores in the PDMS material were filled with water molecules.

[0074] Experimental Group 6: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, where the temperature and humidity of the incubator were controlled at 37° C. and 90% RH, respectively, without airflow.

[0075] Experimental Group 7: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, with the temperature of the incubator controlled at 37°C and the humidity at 100% RH; an external air line was attached to the surface of the chip to blow from one long side to the other, and the pressure gauge controlled the flow rate to be: ~0.05m / s.

[0076] Experimental Group 8: The cell microfluidic chip 4 was placed in a conventional incubator known in the prior art, where the temperature and humidity of the incubator were controlled at 37° C. and 100% RH, respectively, without airflow.

[0077] Experimental group 9: The long-term cell culture device based on the microfluidic chip of the embodiment was used, so that the cell microfluidic chip 4 was completely immersed in sterile water, and the temperature was controlled at 37°C.

[0078] like Figure 5 As shown, during a period of nearly 70 hours, the evaporation amount of liquid in the channel of the same microfluidic chip under different conditions was statistically analyzed. In the device of the present invention (experimental group 9), the evaporation amount was close to that of experimental group 8, and was much smaller than that of other conditions. Under the premise of suppressing the evaporation of liquid in 4 channels of the cell microfluidic chip, the present invention has the advantages of small size and low cost compared with the existing commercial culture system.

Claims

1. A long-term cell culture device based on a microfluidic chip, comprising a placement rack (1), characterized in that: A culture box (2) is arranged on the top surface of the placement rack (1), and a sample box rack (3) is arranged below the culture box (2); The culture box (2) comprises a conductive heat transfer bottom plate (201), a cell microfluidic chip (4) is fixedly arranged in the middle of the conductive heat transfer bottom plate (201), four transparent side plates (202) are arranged along the vertical direction on the conductive heat transfer bottom plate (201) around the cell microfluidic chip (4), the four transparent side plates (202) are arranged symmetrically in pairs, and a transparent top plate (203) is detachably arranged on the top of the four transparent side plates (202); four transparent side plates (202) are arranged along the vertical direction on the top surface of the conductive heat transfer bottom plate (201); the space enclosed by the conductive heat transfer bottom plate (201), the four transparent side plates (202) and the transparent top plate (203) is a sterile water tank, sterile water is contained in the sterile water tank, and the cell microfluidic chip (4) is completely immersed in the sterile water; The sample box rack (3) comprises a sample box rack bottom plate (301), four sample box rack side plates (302) are fixedly arranged on the top surface of the sample box rack bottom plate (301) along the vertical direction, the four sample box rack side plates (302) are symmetrically arranged in pairs, and a sample box rack top plate (303) is detachably arranged at the top of the four sample box rack side plates (302); a plurality of sample tube placement openings (304) are respectively opened on the lateral sides of the sample box rack top plate (303), the sample tube placement openings (304) are located on the lateral outer sides of the sample box rack side plates (302), a sample tube (5) is arranged in the sample tube placement opening (304), and the liquid outlet end of the sample tube (5) is connected to the liquid inlet end of the cell microfluidic chip (4); A temperature control chip (6) is arranged on the top plate (303) of the sample box rack; a conductive metal sheet (7) is arranged on each of the lateral sides of the conductive heat transfer bottom plate (201); a thermistor (8) is fixedly arranged on the surface of the conductive heat transfer bottom plate (201), and the thermistor (8) is close to the cell microfluidic chip (4); the signal input end of the temperature control chip (6) is connected to the signal output end of the thermistor (8), and the signal output end of the temperature control chip (6) is connected to the conductive metal sheet (7); the space enclosed by the sample box rack top plate (303), the four sample box rack side plates (302) and the sample box rack bottom plate (301) is a battery compartment, and a battery (9) is arranged in the battery compartment; the current output end of the battery (9) is connected to the current input end of the temperature control chip (6).

2. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: The sample box rack (3) has a length of 9 cm, a width of 7 cm and a height of 2.7 cm; the culture box (2) has a length of 11 cm, a width of 7 cm and a height of 2 cm; the placement rack (1) has a length of 16 cm, a width of 12 cm and a height of 7 cm.

3. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: Two current output interfaces (305) and a temperature detection interface (306) are arranged on the sample box rack side plate (302) located on the longitudinal front side; a first wire (10) is arranged in the current output interface (305), and the temperature control chip (6) is connected to the conductive metal sheet (7) through the first wire (10); a second wire (11) is arranged in the temperature detection interface (306), and the temperature control chip (6) is connected to the thermistor (8) through the second wire (11).

4. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: Temperature control chip switches (307) are provided on the sample box rack side plates (302) located at the front and rear in the longitudinal direction.

5. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: A charging interface (308) is provided on the sample box frame side panels (302) located at the front and rear in the longitudinal direction, and the current input end of the battery (9) is connected to the charging interface (308).

6. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: A culture box window (204) is respectively provided on both lateral sides of the transparent top plate (203), and a liquid infusion tube (12) and a gas infusion tube (13) are arranged in the culture box window (204); the sample tube (5) is connected to the cell microfluidic chip (4) through the liquid infusion tube (12), and the gas inlet end of the sample tube (5) is connected to the gas outlet end of the gas infusion tube (13).

7. The long-term cell culture device based on a microfluidic chip according to claim 6, characterized in that: The sample tube (5) comprises a sample tube body (501), the four sides and the bottom of the sample tube body (501) are closed, the top of the sample tube body (501) is open, and a sample tube cover (502) is detachably mounted on the open top of the sample tube body (501), and the sample tube body (501) is used to contain cell fluid.

8. The long-term cell culture device based on a microfluidic chip according to claim 7, characterized in that: The sample tube cover (502) is provided with an infusion needle (503) and a gas delivery needle (504); the bottom end of the infusion needle (503) extends into the cell fluid, the top end of the infusion needle (503) passes through and extends out of the sample tube cover (502), and the infusion needle (503) located above the sample tube cover (502) is connected to the cell microfluidic chip (4) through the infusion tube (12); the bottom end of the gas delivery needle (504) extends out of the sample tube body (501), the top end of the gas delivery needle (504) passes through and extends out of the sample tube cover (502), and the gas delivery needle (504) located above the sample tube cover (502) is connected to the gas delivery tube (13).

9. The long-term cell culture device based on a microfluidic chip according to claim 1, characterized in that: The placement rack (1) comprises a placement rack top plate (101) and a placement rack bottom plate (102), a plurality of support legs (103) are installed between the placement rack top plate (101) and the placement rack bottom plate (102), and the support legs (103) are arranged along the vertical direction; a culture box (2) is arranged on the top surface of the placement rack top plate (101), and a sample box rack (3) is arranged on the top surface of the placement rack bottom plate (102).

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