A microfluidic device for tumor organoid culture and drug experiment and its use method

Abstract

The present invention provides a microfluidic device for tumor organoid culture and drug experiments and a method of use, belonging to the technical field of microfluidic devices. It includes a main body and two rotating shafts, the two rotating shafts are respectively rotatably connected to the top of the main body, and also includes a placement mechanism, the placement mechanism is arranged on the top of the two rotating shafts, and the placement mechanism includes two L-shaped frames. The present invention arranges the first microfluidic chip and the second microfluidic chip to be connected through a connecting tube, thereby reducing the phenomenon of the connection between the first microfluidic chip and the second microfluidic chip being loosened due to twisting, stretching or squeezing during long-term use of the connecting tube, and at the same time improves the stability of the connection between the first microfluidic chip and the second microfluidic chip, thereby reducing the loss of experimental samples and reagents due to internal liquid leakage between the first microfluidic chip and the second microfluidic chip, and the probability of contamination of the entire experimental device and the environment.

Description

Technical Field

[0001] The present invention relates to the technical field of microfluidic devices, and in particular to a microfluidic device for tumor organoid culture and drug experiments and a use method thereof. Background Art

[0002] With the development of cell biology and organoid technology, tumor organoids obtained by culturing tumor cells from cancer patient lesions have become a powerful in vitro model for evaluating tumor precision treatment and personalized medicine. These tumor organoids better maintain the physiological and structural characteristics of the original tumor lesions, can self-renew and proliferate for a long time, and maintain the stability of genetic properties, which are advantages that traditional two-dimensional culture systems cannot achieve.

[0003] The patent with announcement number CN112608841B discloses a microfluidic system for tumor organoid culture and drug experiments and a method of use. The system includes a first external pump, a first external pump, a first microfluidic chip for generating a drug concentration gradient, a second microfluidic chip for organoid culture and a centrifuge tube for collecting waste liquid. The first external pump and the first external pump are respectively connected to the input end of the first microfluidic chip through a hose, and the first microfluidic chip, the second microfluidic chip, and the centrifuge tube are connected in sequence through a hose; the microfluidic system of the invention can controllably perfuse culture medium or drugs into tumor organoids, while adjusting the concentration of the drug and its action time on the tumor organoids, real-time regulating the liquid environment of the tumor organoids, realizing batch culture of tumor organoids and conducting drug efficacy testing experiments, and can observe the growth of tumor organoids, drug response, etc. in real time and continuously by cooperating with a microscope.

[0004] Two microfluidic chips need to be connected by multiple hoses during use. In the prior art, two microfluidic chips are mostly connected by multiple hoses and then directly placed on a desktop or a rack. The two microfluidic chips can move independently. During long-term use, the multiple hoses may be twisted, stretched or squeezed, which may cause the hoses at the connection between the two microfluidic chips to loosen and cause liquid leakage. This not only leads to low stability of the connection between the two microfluidic chips, but also causes loss of experimental samples and reagents and pollution of the entire experimental device and environment due to liquid leakage. Therefore, the present application provides a microfluidic device and a method of use for tumor organoid culture and drug experiments to meet the needs. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a microfluidic device for tumor organoid culture and drug experiments and a method of use to solve the problem that multiple existing hoses are twisted, stretched or squeezed during long-term use, causing the hoses at the connection between two microfluidic chips to loosen and cause liquid leakage, which not only leads to low stability of the connection between the two microfluidic chips, but also causes loss of experimental samples and reagents and pollution of the entire experimental device and the environment due to liquid leakage.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0007] A microfluidic device for tumor organoid culture and drug experiment, comprising a main body and two rotating shaft columns, the two rotating shaft columns are respectively rotatably connected to the top of the main body, and also comprising: a placement mechanism, the placement mechanism is arranged on the top of the two rotating shaft columns, the placement mechanism comprises two L-shaped frames, the two L-shaped frames are respectively fixedly connected to the top of the two rotating shaft columns, a first placement frame is fixedly connected to one end of the two L-shaped frames away from the rotating shaft column, a second placement frame is fixedly connected to one end of the two L-shaped frames close to the rotating shaft column, a fixed frame 1 is fixedly connected to one side of the two L-shaped frames close to each other, a first insertion strip is fixedly connected to one side of the two fixed frames close to the first placement frame, and a second placement frame is fixedly connected to one side of the second placement frame close to the first placement frame. Two fixing frames 2 are connected, a second insertion strip is fixedly connected to one side of the two fixing frames 2 close to the second placement frame, and a plurality of connecting tubes are fixedly connected between the first insertion strip and the second insertion strip; a microfluidic mechanism, the microfluidic mechanism is arranged between the two L-shaped frames, the microfluidic mechanism includes two mounting base plates, the two mounting base plates are respectively slidably connected to the inside of the first placement frame and the second placement frame, a first microfluidic chip is attached to the top of the mounting base plate inside the first placement frame, a second microfluidic chip is attached to the top of the mounting base plate inside the second placement frame, a cover glass is attached to the top of the first microfluidic chip and the second microfluidic chip, and a relative connecting port is opened on the side where the first microfluidic chip and the second microfluidic chip are close to each other.

[0008] Optionally, two movable grooves are provided at one end of the top of the main body away from the rotating shaft column, and two limit grooves are provided at one end of the top of the main body close to the movable groove. A rotating hole connecting block is fixedly connected to the top of the main body between the two rotating shaft columns, and a local screw rod is rotatably connected inside the rotating hole connecting block, and a thread is provided on the outer wall of the local screw rod close to the movable groove.

[0009] Optionally, the two L-shaped frames are symmetrically arranged, and the placement mechanism also includes two strip sliding holes, which are respectively opened at the bottom of the two L-shaped frames, and the ends of the two L-shaped frames away from the rotating shaft column are fixedly connected to the extension connecting strip, and the bottoms of the two extension connecting strips are fixedly connected to the limited position abutment strips, and the ends of the bottoms of the two L-shaped frames away from the rotating shaft column are fixedly connected to the rotating wheel support frames, and the sides of the two rotating wheel support frames close to each other are rotatably connected to pulleys.

[0010] Optionally, an injection mechanism is provided on the inner wall at the bottom of the L-shaped frame, and the injection mechanism includes a syringe, which is fixedly connected to one end of the inner wall at the bottom of the L-shaped frame away from the rotating shaft column, and an external pump connection end is fixedly connected to the outer wall of the syringe, and the external pump connection end is connected to the external pump, and a propulsion sealing column is slidably connected inside the syringe.

[0011] Optionally, a penetrating rod is fixedly connected to the side of the pushing sealing column close to the rotating shaft column, and the penetrating rod slides through the side of the syringe away from the pushing sealing column, and an insertion strip is fixedly connected to the end of the outer wall of the penetrating rod away from the pushing sealing column, and the insertion strip is distributed at the bottom of the L-shaped frame, and the connection between the penetrating rod and the insertion strip is slidingly connected to the inside of the strip sliding hole, and an insertion rod is slidingly connected to the bottom of the L-shaped frame, and the insertion rod slides through the bottom of the L-shaped frame, and the insertion rod is plugged into the socket on the insertion strip, and an upper strip plate is fixedly connected to the top of the insertion rod, and an inclined push frame is fixedly connected to the end of the bottom of the upper strip plate away from the insertion rod, and the connection between the inclined push frame and the upper strip plate slides through the bottom of the L-shaped frame.

[0012] Optionally, an elastic spring is fixedly connected between the bottom of the upper strip and the bottom inner wall of the L-shaped frame, the bottom of the inclined push frame is an inclined surface, a push rod is slidably connected to the side of the limit push bar close to the syringe, the push rod slides through the limit push bar, and an offset plate is fixedly connected to the side of the push rod away from the limit push bar.

[0013] Optionally, a tension spring is fixedly connected between the offset plate and the limit push bar, an extension rod is fixedly connected to the side of the offset plate away from the push rod, a push rod is fixedly connected to the side of the extension rod away from the offset plate, the outer wall of the push rod is abutted against the inclined surface at the bottom of the inclined push frame, and there are two injection mechanisms, which are respectively arranged corresponding to the two L-shaped frames.

[0014] Optionally, an adjustment mechanism is provided on the top of the main body, and the adjustment mechanism includes a screw sleeve frame, which is threadedly connected to the outer wall of a local screw, and the outer wall of the screw sleeve frame is fixedly connected to two tripods, and the two tripods are respectively slidably connected to the inside of two movable slide grooves, and the two extended connecting bars are rotatably connected to the inside of vertical movable bars, and the bottoms of the two vertical movable bars are fixedly connected to weight blocks, and the sides of the two weight blocks that are close to each other are rotatably connected to the limiting movable bar, and the ends of the two limiting movable bars away from the weight blocks are respectively movably connected to the inside of the limiting slide grooves.

[0015] Optionally, the microfluidic mechanism also includes a discharge port, which is opened at one end of the bottom of the second microfluidic chip away from the relative connection port, and a waste liquid collection box is slidably connected to one end of the two L-shaped frames away from the first placement rack. The waste liquid collection box is arranged at the bottom of the second placement rack, and the waste liquid collection box is arranged corresponding to the discharge port.

[0016] A method for using a microfluidic device for tumor organoid culture and drug experiments, comprising the following steps:

[0017] S1: injecting the buffer liquid and the culture medium or the drug into two syringes respectively, and connecting the two syringes to two external pumps through two external pump connection ends respectively;

[0018] S2: After the microfluidic mechanism is installed on the first placement rack and the second placement rack, and the first microfluidic chip and the second microfluidic chip are connected through the first insertion strip and the connecting tube and the second insertion strip, the injection tubes at the output ends of the two syringes are respectively connected to the two liquid inlet connection ports on the side of the first microfluidic chip away from the opposite connection port;

[0019] S3: After setting parameters of two external pumps in advance according to experimental requirements to accurately adjust the flow rate, respectively control the thrust of two propulsion sealing columns inside the syringe to control the injection of buffer liquid and culture medium or drug into the first microfluidic chip to perform experiments;

[0020] S4: When the first microfluidic chip and the second microfluidic chip are respectively mounted on the first placement rack and the second placement rack, the mounting base plate and the cover glass are respectively adhered to the bottom and top of the first microfluidic chip, and at the same time, the cover glass is arranged on the top of the first microfluidic chip and the cover glass, so that the state of cells inside the first microfluidic chip and the cover glass can be observed through a microscope.

[0021] Compared with the prior art, the present invention has at least the following beneficial effects:

[0022] In the above scheme, the first insertion strip and the second insertion strip are respectively fixed between the two L-shaped frames by the fixing frame one and the fixing frame two. When the first microfluidic chip and the second microfluidic chip are installed on the first placement frame and the second placement frame, the first microfluidic chip can be directly pushed to slide in the direction of the first insertion strip, and the second microfluidic chip can be pushed to slide to one side of the second insertion strip, so that the relative connection ports on the first microfluidic chip and the second microfluidic chip are respectively connected to the first insertion strip and the second insertion strip, so that the first microfluidic chip and the second microfluidic chip are connected through the connecting tube, reducing the phenomenon of the connection between the first microfluidic chip and the second microfluidic chip loosening due to twisting, stretching or squeezing during long-term use, causing liquid leakage, and at the same time improving the stability of the connection between the first microfluidic chip and the second microfluidic chip, thereby reducing the loss of experimental samples and reagents due to liquid leakage inside the first microfluidic chip and the second microfluidic chip, and the probability of pollution to the entire experimental device and the environment.

[0023] The bottom of the main body is fitted with the desktop so that the device can tilt with the desktop, and the vertical movable bar is tilted inside the extended connecting bar by the weight of the load-bearing block, so that the vertical movable bar and the load-bearing block are perpendicular to the ground, and the local screw rod is manually controlled to rotate inside the rotating hole connecting block, and the screw rod sleeve is threadedly connected to the local screw rod, which will drive the two tripods to move forward in the two moving slide grooves respectively. At the same time, the pulleys on the two rotating wheel support frames are respectively against the inclined surfaces of the two tripods, which will gradually push the two L-shaped frames as a whole to rotate at one end of the top of the main body away from the moving slide groove through the two rotating shafts, so that the angle between the L-shaped frame and the main body can be increased, and the placement mechanism that follows the tilt of the desktop can be adjusted to a horizontal posture, thereby avoiding the desktop being tilted. The angular deviation causes the microfluidic mechanisms on the first placement rack and the second placement rack to tilt synchronously, and when the desktop is tilted, the vertical movable bar will rotate inside the extended connecting bar through the gravity applied by the weight block, and the center of gravity of the vertical movable bar is always in a vertical downward posture, so that the deviation angle between the vertical movable bar and the limit bar is generated, which makes it convenient for the user to observe whether the desktop is tilted, and then adjust the posture of the microfluidic mechanism inside the placement mechanism to make it horizontal according to the deviation angle between the vertical movable bar and the limit bar, thereby reducing the occurrence of liquid flow disorder, uneven liquid distribution, bubble generation and aggregation due to the tilt of the microfluidic mechanism, so that the experiment can be carried out smoothly and the accuracy of the experimental results is improved.

[0024] When the L-shaped frame is in a horizontal position by rotating the shaft column, the weight block and the vertical movable bar gradually approach the limit stop bar. When the L-shaped frame gradually becomes horizontal through continuous rotation of the L-shaped frame, the vertical movable bar will squeeze the stop bar and make the vertical movable bar stick to the limit stop bar. When the vertical movable bar gradually sticks to the limit stop bar, the two ends of the limit movable bar are respectively connected with the weight block and the inside of the limit slide groove. When the vertical movable bar and the limit stop bar stick together, the limit movable bar limits the tripod to stop sliding in the moving slide groove, and the local lead screw cannot rotate either. Therefore, during the process of adjusting the angle of the L-shaped frame, when the vertical movable bar sticks to the limit stop bar and the local lead screw cannot rotate, the placement mechanism and the microfluidic mechanism will be exactly in a horizontal position, thereby improving the accuracy of adjusting the placement mechanism to a horizontal position, avoiding the adjusted angle between the L-shaped frame and the main body being greater than the inclination angle of the seat surface, and improving the convenience of the device to adjust the L-shaped frame to a horizontal state.

[0025] When the lever is in the air, the spring will not move, and the cylinder will move upwards as the lever moves, causing the cylinder to move in an arcuate manner, thereby releasing the cylinder from the engagement of the cylinder and causing the cylinder to move in an arcuate manner. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the invention and, together with the description, further serve to explain the principles of the invention and to enable those skilled in the relevant art to make and use the invention.

[0027] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0028] Figure 2 This is a schematic diagram of the present invention in which the microfluidic mechanism is removed as a whole;

[0029] Figure 3 It is a schematic diagram of the main structure of the present invention;

[0030] Figure 4 It is a schematic diagram of the structure of the liquid injection mechanism of the present invention;

[0031] Figure 5 This is a schematic diagram of the bottom structure of the placement mechanism of the present invention;

[0032] Figure 6 It is a schematic diagram of the structure of the regulating mechanism of the present invention;

[0033] Figure 7 It is a schematic diagram of the locking mechanism structure of the present invention;

[0034] Figure 8 for Figure 3 Enlarged view of point A in the middle;

[0035] Fig. 9 Figure 2 is a diagram of the method of the present invention.

[0036] Reference numerals:

[0037] 1. Main body; 101. Moving slide; 102. Limiting slide; 103. Rotating hole connection block; 104. Local screw rod; 105. Rotating shaft column; 2. Placing mechanism; 201. L-shaped frame; 202. First placing frame; 203. Second placing frame; 204. Fixed frame one; 205. First insertion strip; 206. Connecting pipe; 207. Second insertion strip; 208. Fixed frame two; 209. Strip slide hole; 210. Extended connecting strip; 211. Limiting strip; 212. Rotating wheel support frame; 3. Injection mechanism; 301. Syringe; 302. External pump connection end; 303. Push Sealing column; 304, through rod; 305, jack strip; 306, insertion rod; 307, upper strip plate; 308, inclined push frame; 309, extension rod; 310, offset plate; 311, support rod; 312, push rod; 4, adjustment mechanism; 401, screw sleeve; 402, tripod; 403, vertical movable strip; 404, weight block; 405, limit movable strip; 5, microfluidic mechanism; 501, mounting base plate; 502, first microfluidic chip; 503, cover glass; 504, relative connection port; 505, second microfluidic chip; 506, discharge port; 6, waste liquid collection box.

[0038] As shown in the figure, in order to clearly implement the structure of the embodiment of the present invention, specific structures and devices are marked in the figure, but this is only for illustrative purposes and is not intended to limit the present invention to the specific structure, device and environment. According to specific needs, ordinary technicians in this field can adjust or modify these devices and environments. DETAILED DESCRIPTION

[0039] The following is a detailed description of a microfluidic device and a method of use for tumor organoid culture and drug experiments provided by the present invention in conjunction with the accompanying drawings and specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternative methods to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.

[0040] It should be noted that the references to "one embodiment", "embodiment", "exemplary embodiments", "some embodiments" and the like in the specification indicate that the embodiments described may include specific features, structures or characteristics, but not every embodiment may include the specific features, structures or characteristics. In addition, when a specific feature, structure or characteristic is described in conjunction with an embodiment, it should be within the knowledge of a person skilled in the art to implement such feature, structure or characteristic in conjunction with other embodiments (whether or not explicitly described).

[0041] In general, a term can be understood, at least in part, from its use in context. For example, depending, at least in part, on the context, the term "one or more" as used herein can be used to describe any feature, structure, or characteristic in the singular sense, or can be used to describe a combination of features, structures, or characteristics in the plural sense. Additionally, the term "based on" can be understood as not necessarily intended to convey an exclusive set of factors, but can instead, depending, at least in part, on the context, allow for the presence of other factors that are not necessarily explicitly described.

[0042] It will be understood that the meanings of “on,” “over,” and “above” in the present invention should be interpreted in the broadest manner, so that “on” not only means “directly on” something, but also includes the meaning of being “on” something with intervening features or layers therebetween, and “on” or “over” not only means “on” or “above” something, but also includes the meaning of being “on” or “above” something with no intervening features or layers therebetween.

[0043] Additionally, spatially relative terms such as "under," "beneath," "lower," "above," "upper," and the like may be used herein for descriptive convenience to describe the relationship of one element or feature to another element or features, as shown in the accompanying drawings. Spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the accompanying drawings. The device may be oriented in other ways, and the spatially relative descriptors used herein may be similarly interpreted accordingly.

[0044] like Figures 1 to 3 , Figure 5 and Figure 6As shown, an embodiment of the present invention provides a microfluidic device for tumor organoid culture and drug experiments and a method of use, including a main body 1 and two rotating shafts 105, the two rotating shafts 105 are respectively rotatably connected to the top of the main body 1, two movable slide grooves 101 are provided at one end of the top of the main body 1 away from the rotating shaft 105, and two limiting slide grooves 102 are provided at one end of the top of the main body 1 close to the movable slide groove 101, and a rotating hole connecting block 103 is fixedly connected at a place on the top of the main body 1 between the two rotating shafts 105, and a local screw rod 104 is rotatably connected inside the rotating hole connecting block 103, and a thread is provided on the outer wall of the local screw rod 104 close to the movable slide groove 101.

[0045] like Figures 1 to 3 , Figure 5 and Figure 8 As shown, it includes a placement mechanism 2, which is arranged on the top of two rotating shaft columns 105. The placement mechanism 2 includes two L-shaped frames 201, and the two L-shaped frames 201 are respectively fixedly connected to the top of the two rotating shaft columns 105. The end between the two L-shaped frames 201 away from the rotating shaft column 105 is fixedly connected to a first placement frame 202, and the end between the two L-shaped frames 201 close to the rotating shaft column 105 is fixedly connected to a second placement frame 203, and the sides of the two L-shaped frames 201 close to each other are fixedly connected to a fixed frame 1 204, and the sides of the two fixed frames 1 204 close to the first placement frame 202 are fixedly connected to a first insertion strip 205, and the side of the second placement frame 203 close to the first placement frame 202 is fixedly connected to two fixed frames 208, and the side of the two fixed frames 208 close to the second placement frame 203 is fixedly connected to a second insertion strip 20 7. A plurality of connecting tubes 206 are fixedly connected between the first insertion strip 205 and the second insertion strip 207. The first insertion strip 205 and the second insertion strip 207 are fixed between the two L-shaped frames 201 through the fixing frame 1 204 and the fixing frame 2 208, respectively. When the first microfluidic chip 502 and the second microfluidic chip 505 are installed on the first placement frame 202 and the second placement frame 203, the first microfluidic chip 502 can be directly pushed to slide in the direction of the first insertion strip 205, and the second microfluidic chip 505 can be pushed to slide to one side of the second insertion strip 207, so that the relative connecting ports 504 on the first microfluidic chip 502 and the second microfluidic chip 505 are respectively connected to the first insertion strip 205 and the second insertion strip 207, so that the first microfluidic chip 502 and the second microfluidic chip 505 are connected through the connecting tubes 206.

[0046] like Figures 1 to 3 , Figure 5 and Figure 8As shown, the two L-shaped frames 201 are symmetrically arranged, and the placement mechanism 2 also includes two strip sliding holes 209, and the two strip sliding holes 209 are respectively opened at the bottom of the two L-shaped frames 201, and the ends of the two L-shaped frames 201 away from the rotating shaft column 105 are fixedly connected with the extension connecting strip 210, and the bottoms of the two extension connecting strips 210 are fixedly connected with the limited position abutting strip 211, and the ends of the bottoms of the two L-shaped frames 201 away from the rotating shaft column 105 are fixedly connected with the rotating wheel support frame 212, and the sides of the two rotating wheel support frames 212 close to each other are rotatably connected with pulleys. When the L-shaped frame 201 is rotated by the rotating shaft column 105 to make the L-shaped frame 201 horizontal, the load block 404 and the vertical movable bar 403 gradually approach the limited position abutting strip 211. When the L-shaped frame 201 is continuously rotated, the load block 404 and the vertical movable bar 403 gradually move closer to the limited position abutting strip 211. After the rotation gradually becomes horizontal, the vertical movable bar 403 will squeeze the support rod 311 and make the vertical movable bar 403 stick to the limit support bar 211. When the vertical movable bar 403 gradually sticks to the limit support bar 211, the two ends of the limit movable bar 405 are respectively connected with the load block 404 and the inside of the limit slide 102. When the vertical movable bar 403 sticks to the limit support bar 211, the tripod 402 can stop sliding in the movable slide 101 through the limit of the limit movable bar 405, and the local screw rod 104 cannot rotate. Therefore, during the process of adjusting the angle of the L-shaped frame 201, when the vertical movable bar 403 sticks to the limit support bar 211 and the local screw rod 104 cannot rotate, the placement mechanism 2 and the microfluidic mechanism 5 will be in a horizontal posture;

[0047] like Figures 1 to 3 , Figure 5As shown, the microfluidic mechanism 5 includes a microfluidic mechanism 5, which is arranged between two L-shaped frames 201. The microfluidic mechanism 5 includes two mounting bases 501, which are respectively slidably connected to the inside of the first placement frame 202 and the second placement frame 203. The top of the mounting base 501 inside the first placement frame 202 is attached to the first microfluidic chip 502, and the top of the mounting base 501 inside the second placement frame 203 is attached to the second microfluidic chip 505. The tops of the first microfluidic chip 502 and the second microfluidic chip 505 are both attached to the cover glass 503. The first microfluidic chip 502 and the second microfluidic chip 505 are both provided with a relative connection port 504 on the side where the first microfluidic chip 502 and the second microfluidic chip 505 are close to each other. The microfluidic mechanism 5 also includes a discharge port 506, which is opened at one end of the bottom of the second microfluidic chip 505 away from the relative connection port 504, and a waste liquid collection box 6 is slidably connected to one end of the two L-shaped frames 201 away from the first placement frame 202. The liquid collection box 6 is arranged at the bottom of the second placement rack 203, and the waste liquid collection box 6 is arranged corresponding to the discharge port 506. The buffer liquid and the culture medium or the drug are respectively injected into the two syringes 301, and the two syringes 301 are respectively connected to the two external pumps through the two external pump connection ends 302. After the microfluidic mechanism 5 is installed on the first placement rack 202 and the second placement rack 203, and the first microfluidic chip 502 and the second microfluidic chip 505 are connected through the first insertion strip 205 and the connecting tube 206 and the second insertion strip 207, the injection tubes at the output ends of the two syringes 301 are respectively connected to the two liquid inlet connection ports of the first microfluidic chip 502 away from the relative connection port 504. After the two external pumps are set with parameters in advance according to the experimental requirements to accurately adjust the flow rate, the thrust of the two propulsion sealing columns 303 in the syringe 301 is respectively controlled to control the buffer liquid and the culture medium or the drug to be injected into the first microfluidic chip 502 for experiments;

[0048] When the first microfluidic chip 502 and the second microfluidic chip 505 are respectively mounted on the first placement rack 202 and the second placement rack 203, the mounting base 501 and the cover glass 503 are respectively adhered to the bottom and top of the first microfluidic chip 502, and the cover glass 503 is arranged on the top of the first microfluidic chip 502 and the cover glass 503, so as to facilitate the observation of the state of cells inside the first microfluidic chip 502 and the cover glass 503 through a microscope.

[0049] When in use, the first insertion strip 205 and the second insertion strip 207 are fixed between the two L-shaped frames 201 through the fixing frame 1 204 and the fixing frame 2 208 respectively, and when the first microfluidic chip 502 and the second microfluidic chip 505 are installed on the first placement frame 202 and the second placement frame 203, the first microfluidic chip 502 can be directly pushed to slide in the direction of the first insertion strip 205, and the second microfluidic chip 505 can be pushed to slide to one side of the second insertion strip 207, so that the first microfluidic chip 502 and the second microfluidic chip 505 can be directly pushed to slide in the direction of the first insertion strip 205, and the second microfluidic chip 505 can be pushed to slide to one side of the second insertion strip 207, so that the first microfluidic chip 502 and the second microfluidic chip 505 can be directly pushed to slide in the direction of the first insertion strip 205, and the second microfluidic chip 505 can be pushed to slide to the one side of the second insertion strip 207, so that the first microfluidic chip 502 and the second microfluidic chip 505 can be directly pushed to slide in the direction of the first insertion strip 205, ... first microfluidic chip The relative connection ports 504 on the microfluidic chip 505 are respectively connected to the first insertion strip 205 and the second insertion strip 207, so that the first microfluidic chip 502 and the second microfluidic chip 505 are connected through the connecting tube 206, which reduces the phenomenon of the connection between the first microfluidic chip 502 and the second microfluidic chip 505 being loosened due to twisting, stretching or squeezing during long-term use, thereby improving the stability of the connection between the first microfluidic chip 502 and the second microfluidic chip 505;

[0050] like Figure 1 , Figure 2 , Figure 5 and Figure 7As shown, the inner wall at the bottom of the L-shaped frame 201 is provided with an injection mechanism 3, and the injection mechanism 3 includes a syringe 301, and the syringe 301 is fixedly connected to the end of the inner wall at the bottom of the L-shaped frame 201 away from the rotating shaft column 105, and the outer wall of the syringe 301 is fixedly connected to the external pump connection end 302, and the external pump connection end 302 is connected to the external pump, and the syringe 301 is slidably connected to a propulsion sealing column 303, and a side of the propulsion sealing column 303 close to the rotating shaft column 105 is fixedly connected to a through rod 304, and the through rod 304 slides through the side of the syringe 301 away from the propulsion sealing column 303, and the end of the outer wall of the through rod 304 away from the propulsion sealing column 303 is fixedly connected to a socket strip 305, and the socket strips 305 are distributed at the bottom of the L-shaped frame 201, and the through rod 304 is fixedly connected to the end of the outer wall of the through rod 304 away from the propulsion sealing column 303. The connection between the penetration rod 304 and the socket strip 305 is slidably connected with the inside of the strip sliding hole 209, and the bottom of the L-shaped frame 201 is slidably connected with an insertion rod 306, which slides through the bottom of the L-shaped frame 201, and the insertion rod 306 is plugged into the socket on the socket strip 305. The top of the insertion rod 306 is fixedly connected with an upper strip plate 307, and the end of the bottom of the upper strip plate 307 away from the insertion rod 306 is fixedly connected with an inclined push frame 308, and the connection between the inclined push frame 308 and the upper strip plate 307 slides through the bottom of the L-shaped frame 201, and an elastic spring is fixedly connected between the bottom of the upper strip plate 307 and the bottom inner wall of the L-shaped frame 201, and the bottom of the inclined push frame 308 is an inclined surface, and the side of the limit stop strip 211 close to the syringe 301 is slidably connected with a stop rod 311, which stops The rod 311 slides through the limiting push bar 211, and the side of the push bar 311 away from the limiting push bar 211 is fixedly connected with a deflection plate 310, and a tension spring is fixedly connected between the deflection plate 310 and the limiting push bar 211, and the side of the deflection plate 310 away from the push bar 311 is fixedly connected with an extension rod 309, and the side of the extension rod 309 away from the deflection plate 310 is fixedly connected with a push rod 312, and the outer wall of the push rod 312 is against the inclined surface at the bottom of the inclined push frame 308. There are two injection mechanisms 3, and the two injection mechanisms 3 are respectively arranged corresponding to the two L-shaped frames 201. When the vertical movable bar 403 contacts the push bar 311, the vertical movable bar 403 can squeeze the push bar 311, so that the tension spring between the deflection plate 310 and the limiting push bar 211 is pulled. The push rod 312 is extended and the extension rod 309 is pushed to move backward. The push rod 312 is against the inclined surface of the inclined push frame 308, which pushes the round rod on the top of the inclined push frame 308 to slide through the bottom of the L-shaped frame 201, driving the upper strip 307 and the insertion rod 306 to slide upward synchronously. When the vertical movable bar 403 contacts the limiting abutting bar 211, the insertion rod 306 is just separated from the socket on the socket bar 305, so that the socket bar 305 is separated from the limit of the insertion rod 306. When the L-shaped frame 201 is not in a horizontal posture, the insertion rod 306 is inserted into the socket bar 305 to limit it. When the external pump applies a thrust to the propulsion sealing column 303 inside the syringe 301, the propulsion sealing column 303 cannot be pushed to move inside the syringe 301.

[0051] like Figure 1 , Figure 2 , Figures 5 to 7 As shown, an adjustment mechanism 4 is provided on the top of the main body 1, and the adjustment mechanism 4 includes a screw sleeve 401, the screw sleeve 401 is threadedly connected to the outer wall of the local screw 104, and the outer wall of the screw sleeve 401 is fixedly connected with two tripods 402, and the two tripods 402 are respectively slidably connected to the inside of the two movable slide grooves 101, and the two extended connecting bars 210 are rotatably connected to the inside of the vertical movable bars 403, and the bottoms of the two vertical movable bars 403 are fixedly connected to the weight blocks 404, and the sides of the two weight blocks 404 close to each other are rotatably connected to the limited movable bars 405, and the ends of the two limited movable bars 405 away from the weight blocks 404 are respectively movably connected to the inside of the limited slide grooves 102, so that the device follows the inclination of the desktop through the bottom of the main body 1 fitting with the desktop, and the vertical movable bars 403 are connected through The load of the overload block 404 causes the vertical movable bar 403 to tilt inside the extended connecting bar 210, so that the vertical movable bar 403 and the load block 404 are perpendicular to the ground. The local screw rod 104 is manually controlled to rotate inside the rotary hole connecting block 103, and the screw rod sleeve 401 is threadedly connected to the local screw rod 104, which will drive the two tripods 402 to move forward respectively inside the two movable slide grooves 101. At the same time, the pulleys on the two rotating wheel support frames 212 are respectively against the inclined surfaces of the two tripods 402, which will gradually push the two L-shaped frames 201 as a whole to rotate at one end of the top of the main body 1 away from the movable slide groove 101 through the two rotating shaft columns 105, so that the angle between the L-shaped frame 201 and the main body 1 can be increased, so that the placement mechanism 2 that follows the inclination of the desktop is adjusted to a horizontal posture.

[0052] A method for using a microfluidic device for tumor organoid culture and drug experiments, comprising the following steps:

[0053] S1: injecting the buffer liquid and the culture medium or the drug into the two syringes 301 respectively, and connecting the two syringes 301 to the two external pumps through the two external pump connection ends 302 respectively;

[0054] S2: After the microfluidic mechanism 5 is installed on the first placement rack 202 and the second placement rack 203, and the first microfluidic chip 502 and the second microfluidic chip 505 are connected through the first insertion strip 205, the connecting tube 206 and the second insertion strip 207, the injection tubes at the output ends of the two syringes 301 are respectively connected to the two liquid inlet connection ports on the side of the first microfluidic chip 502 away from the opposite connection port 504;

[0055] S3: After setting parameters of two external pumps in advance according to experimental requirements to accurately adjust the flow rate, respectively control the thrust of the two propulsion sealing columns 303 inside the syringe 301 to control the injection of the buffer liquid and the culture medium or the drug into the first microfluidic chip 502 to perform experiments;

[0056] S4: When the first microfluidic chip 502 and the second microfluidic chip 505 are respectively mounted on the first placement rack 202 and the second placement rack 203, the mounting base 501 and the cover glass 503 are respectively adhered to the bottom and the top of the first microfluidic chip 502. At the same time, the cover glass 503 is arranged on the top of the first microfluidic chip 502 and the cover glass 503, and the state of the cells inside the first microfluidic chip 502 and the cover glass 503 can be observed through a microscope.

[0057] The working principle of the technical solution provided by the present invention is as follows:

[0058] When in use, after the device is placed on a desktop that is not in a horizontal state, the bottom of the main body 1 is fitted with the desktop to make the device tilt with the desktop, and the vertical movable bar 403 is tilted inside the extended connecting bar 210 through the weight of the load-bearing block 404, so that the vertical movable bar 403 and the load-bearing block 404 are vertical to the ground, and the local screw rod 104 is manually controlled to rotate inside the rotary hole connecting block 103, and the screw rod sleeve 401 is threadedly connected to the local screw rod 104, which will drive the two tripods 402 to move forward respectively inside the two movable slide grooves 101, and at the same time, the pulleys on the two rotating wheel support frames 212 are respectively against the inclined surfaces of the two tripods 402, which will gradually push the two L-shaped frames 201 as a whole to move away from the movable slide grooves 101 at the top of the main body 1 through the two rotating shaft columns 105. One end is rotated, so that the angle between the L-shaped frame 201 and the main body 1 is increased, and the placement mechanism 2 that follows the tilt of the desktop is adjusted to a horizontal posture, thereby avoiding the microfluidic mechanism 5 on the first placement frame 202 and the second placement frame 203 from being tilted synchronously due to the angle deviation of the desktop. When the desktop is tilted, the vertical movable bar 403 will rotate inside the extended connecting bar 210 through the gravity applied by the load-bearing block 404, and the center of gravity of the vertical movable bar 403 is always in a vertical downward posture, so that the deviation angle between the vertical movable bar 403 and the limit stop bar 211 is generated, which makes it convenient for the user to observe whether the desktop is tilted, and then adjust the posture of the microfluidic mechanism 5 inside the placement mechanism 2 to make it horizontal according to the deviation angle between the vertical movable bar 403 and the limit stop bar 211;

[0059] When the L-shaped frame 201 is rotated by the rotating shaft column 105 to make the L-shaped frame 201 horizontal, the load block 404 and the vertical movable bar 403 gradually approach the limit stop bar 211. When the L-shaped frame 201 is gradually horizontal through the continuous rotation of the L-shaped frame 201, the vertical movable bar 403 will squeeze the stop bar 311 and make the vertical movable bar 403 and the limit stop bar 211 stick together. When the vertical movable bar 403 gradually sticks to the limit stop bar 211, the two ends of the limit moveable bar 405 are respectively connected to the load block 404. 04 is movably connected with the inside of the limit slide groove 102. When the vertical movable bar 403 is in contact with the limit stop bar 211, the tripod 402 can stop sliding in the movable slide groove 101 through the limit of the limit movable bar 405, and the local screw rod 104 cannot rotate. Therefore, during the process of adjusting the angle of the L-shaped frame 201, when the vertical movable bar 403 is in contact with the limit stop bar 211 and the local screw rod 104 cannot rotate, the placement mechanism 2 and the microfluidic mechanism 5 will be in a horizontal posture;

[0060] When the vertical movable bar 403 approaches the limit push bar 211, the two ends of the limit movable bar 405 are respectively connected with the load block 404 and the limit slide groove 102. When the vertical movable bar 403 contacts the push rod 311, the vertical movable bar 403 can squeeze the push rod 311, so that the tension spring between the offset plate 310 and the limit push bar 211 is stretched, and the extension rod 309 is pushed to move backward. The push rod 312 is against the inclined surface of the inclined push frame 308, which will push the round rod at the top of the inclined push frame 308 to slide through the bottom of the L-shaped frame 201, driving the upper strip 307 and the insertion plate 307 to slide. The rod 306 slides upward synchronously, and when the vertical movable bar 403 contacts the limiting abutment bar 211, the insertion rod 306 is separated from the socket on the socket bar 305, so that the socket bar 305 is free from the limitation of the insertion rod 306. When the L-shaped frame 201 is not in a horizontal posture, the insertion rod 306 is inserted inside the socket bar 305 to limit it. When an external pump applies a thrust to the propulsion sealing column 303 inside the syringe 301, the propulsion sealing column 303 cannot be pushed to move inside the syringe 301, so that the L-shaped frame 201 of the device and the microfluidic mechanism 5 can be operated and used only when they are in a horizontal posture.

[0061] The present invention covers any substitution, modification, equivalent method and scheme made on the essence and scope of the present invention. In order to make the public have a thorough understanding of the present invention, specific details are described in detail in the following preferred embodiments of the present invention, but those skilled in the art can fully understand the present invention without the description of these details. In addition, in order to avoid unnecessary confusion about the essence of the present invention, well-known methods, processes, procedures, components and circuits are not described in detail.

[0062] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A microfluidic device for tumor organoid culture and drug experiments, characterized in that: It comprises a main body and two rotating shaft columns, wherein the two rotating shaft columns are respectively rotatably connected to the top of the main body, and is characterized in that it also comprises: A placing mechanism, wherein the placing mechanism is arranged on the top of two rotating shaft columns, and the placing mechanism includes two L-shaped frames, the two L-shaped frames are respectively fixedly connected to the top of the two rotating shaft columns, a first placing frame is fixedly connected to one end of the two L-shaped frames away from the rotating shaft column, a second placing frame is fixedly connected to one end of the two L-shaped frames close to the rotating shaft column, a fixing frame 1 is fixedly connected to one side of the two L-shaped frames close to each other, a first insertion strip is fixedly connected to one side of the two fixing frames 1 close to the first placing frame, two fixing frames 2 are fixedly connected to one side of the second placing frame close to the first placing frame, a second insertion strip is fixedly connected to one side of the two fixing frames 2 close to the second placing frame, and a plurality of connecting tubes are fixedly connected between the first insertion strip and the second insertion strip; A microfluidic mechanism, the microfluidic mechanism is arranged between two L-shaped frames, the microfluidic mechanism includes two mounting base plates, the two mounting base plates are respectively slidably connected inside a first placement frame and a second placement frame, a first microfluidic chip is attached to the top of the mounting base plate inside the first placement frame, a second microfluidic chip is attached to the top of the mounting base plate inside the second placement frame, a cover glass is attached to the top of the first microfluidic chip and the second microfluidic chip, and a relative connection port is opened on the side where the first microfluidic chip and the second microfluidic chip are close to each other; The two L-shaped frames are symmetrically arranged, and the placement mechanism also includes two strip sliding holes, which are respectively opened at the bottom of the two L-shaped frames, and the ends of the two L-shaped frames away from the rotating shaft column are fixedly connected to the extension connecting strips, and the bottoms of the two extension connecting strips are fixedly connected to the limited position abutment strips, and the ends of the bottoms of the two L-shaped frames away from the rotating shaft column are fixedly connected to the rotating wheel support frames, and the sides of the two rotating wheel support frames close to each other are rotatably connected to the pulleys; Two movable slide grooves are arranged at one end of the top of the main body away from the rotating shaft column, and two limiting slide grooves are arranged at one end of the top of the main body close to the movable slide groove. A rotating hole connecting block is fixedly connected to a place on the top of the main body between the two rotating shaft columns, and a local screw rod is rotatably connected inside the rotating hole connecting block, and a thread is arranged on the end of the outer wall of the local screw rod close to the movable slide groove; An adjustment mechanism is provided on the top of the main body, and the adjustment mechanism includes a screw sleeve, which is threadedly connected to the outer wall of a local screw, and the outer wall of the screw sleeve is fixedly connected to two tripods, and the two tripods are respectively slidably connected to the inside of two movable slide grooves, and the two extended connecting bars are rotatably connected to the inside of vertical movable bars, and the bottoms of the two vertical movable bars are fixedly connected to weight blocks, and the sides of the two weight blocks close to each other are rotatably connected to the limiting movable bar, and the ends of the two limiting movable bars away from the weight blocks are respectively movably connected to the inside of the limiting slide groove.

2. The microfluidic device for tumor organoid culture and drug experiment according to claim 1, characterized in that: The bottom inner wall of the L-shaped frame is provided with an injection mechanism, and the injection mechanism includes a syringe. The syringe is fixedly connected to one end of the bottom inner wall of the L-shaped frame away from the rotating shaft column. The outer wall of the syringe is fixedly connected to an external pump connection end, and the external pump connection end is connected to an external pump. The syringe is slidably connected to a propulsion sealing column.

3. The microfluidic device for tumor organoid culture and drug experiment according to claim 2, characterized in that: The side of the advancement sealing column close to the rotating shaft column is fixedly connected with a through rod, and the through rod slides through the side of the syringe away from the advancement sealing column, and the end of the outer wall of the through rod away from the advancement sealing column is fixedly connected with a socket strip, and the socket strip is distributed at the bottom of the L-shaped frame, and the connection between the through rod and the socket strip is slidably connected with the inside of the strip sliding hole, and the bottom of the L-shaped frame is slidably connected with an insertion rod, and the insertion rod slides through the bottom of the L-shaped frame, and the insertion rod is plugged into the socket on the socket strip, and the top of the insertion rod is fixedly connected with an upper strip plate, and the bottom of the upper strip plate is fixedly connected with an inclined push frame at one end away from the insertion rod, and the connection between the inclined push frame and the upper strip plate slides through the bottom of the L-shaped frame.

4. The microfluidic device for tumor organoid culture and drug experiment according to claim 3, characterized in that: An elastic spring is fixedly connected between the bottom of the upper strip and the bottom inner wall of the L-shaped frame, the bottom of the inclined push frame is an inclined surface, the side of the limit push bar close to the syringe is slidably connected with a push rod, the push rod slides through the limit push bar, and the side of the push rod away from the limit push bar is fixedly connected with an offset plate.

5. The microfluidic device for tumor organoid culture and drug experiment according to claim 4, characterized in that: A tension spring is fixedly connected between the offset plate and the limiting abutment bar, an extension rod is fixedly connected to the side of the offset plate away from the abutment rod, a push rod is fixedly connected to the side of the extension rod away from the offset plate, the outer wall of the push rod abuts against the inclined surface at the bottom of the inclined push frame, and there are two injection mechanisms, which are respectively arranged corresponding to the two L-shaped frames.

6. The microfluidic device for tumor organoid culture and drug experiment according to claim 5, characterized in that: The microfluidic mechanism also includes a discharge port, which is opened at one end of the bottom of the second microfluidic chip away from the relative connection port, and a waste liquid collection box is slidably connected to one end of the two L-shaped frames away from the first placement rack. The waste liquid collection box is arranged at the bottom of the second placement rack, and the waste liquid collection box is arranged corresponding to the discharge port.

7. A method for using a microfluidic device for tumor organoid culture and drug experiment, using the microfluidic device for tumor organoid culture and drug experiment according to claim 6, characterized in that: The following steps are involved: S1: injecting the buffer liquid and the culture medium or the drug into two syringes respectively, and connecting the two syringes to two external pumps through two external pump connection ends respectively; S2: After the microfluidic mechanism is installed on the first placement rack and the second placement rack, and the first microfluidic chip and the second microfluidic chip are connected through the first insertion strip and the connecting tube and the second insertion strip, the injection tubes at the output ends of the two syringes are respectively connected to the two liquid inlet connection ports on the side of the first microfluidic chip away from the opposite connection port; S3: After setting parameters of two external pumps in advance according to experimental requirements to accurately adjust the flow rate, respectively control the thrust of two propulsion sealing columns inside the syringe to control the injection of buffer liquid and culture medium or drug into the first microfluidic chip to perform experiments; S4: When the first microfluidic chip and the second microfluidic chip are respectively mounted on the first placement rack and the second placement rack, the mounting base plate and the cover glass are respectively adhered to the bottom and top of the first microfluidic chip, and at the same time, the cover glass is arranged on the top of the first microfluidic chip and the cover glass, and the state of the cells inside the first microfluidic chip and the cover glass is observed through a microscope.

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