Colon cancer metastasis microfluidic dynamic simulation device suitable for experiment teaching
By designing a microfluidic dynamic simulation device suitable for experimental teaching, and combining a micron-scale pore structure and a piezoelectric micropump, real-time observation and safe operation of the metastasis process of colon cancer cells were realized. This solved the problem of the complexity of dynamic observation and operation of existing devices, and improved the teaching effect and safety.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-26
- Publication Date
- 2026-04-03
AI Technical Summary
Existing colorectal cancer metastasis simulation experimental devices suffer from problems such as inability to dynamically observe the cancer cell migration process, complex operation, poor interactivity, high biosafety risks, and limited mechanism demonstration.
A microfluidic dynamic simulation device for colon cancer metastasis was designed, comprising a microfluidic driving device, a dynamic simulation module, and a teaching imaging system. It uses a capillary and ordinary blood vessel simulation plate with a micron-sized pore structure, combined with a piezoelectric micropump to simulate blood flow, and is equipped with an excitation light source and a microscope camera for real-time observation. It is connected to a biosafety collection bottle to ensure safety.
This method enables real-time observation of the metastasis process of colon cancer cells, reduces operational complexity, improves teaching effectiveness and interactivity, demonstrates richer mechanisms, and ensures biosafety.
Smart Images
Figure CN224082113U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of biomedical teaching experiments, specifically to a microfluidic dynamic simulation device for colon cancer metastasis suitable for experimental teaching. Background Technology
[0002] Colorectal cancer metastasis simulation experiments are a type of experimental method designed to study the mechanisms of colorectal cancer cell metastasis and test potential treatment strategies. In experimental courses for immunology, oncology, and other related majors in higher education institutions, these experiments are often used in conjunction with teaching experimental devices to allow students to observe the colorectal cancer metastasis process more intuitively.
[0003] However, existing teaching experimental devices have the following drawbacks: On the one hand, when using static models such as Transwell chambers or plastic anatomical models, the teaching effect is poor because static models can only show the endpoint of metastasis and cannot dynamically observe the real-time process of cancer cell migration. On the other hand, when using research-grade microfluidic equipment consisting of precision pump control, parameter adjustment, and professional imaging equipment, the high complexity of operation due to the large number of precision instruments makes it difficult for students to master the operation within a limited class time. In addition, there are a series of problems such as poor teaching interactivity, biosafety risks, and limited mechanism demonstration. Summary of the Invention
[0004] To address the shortcomings of existing technologies, the purpose of this invention is to provide a microfluidic dynamic simulation device for colorectal cancer metastasis that allows for real-time observation of the metastasis process and has low operational complexity, making it suitable for experimental teaching and thus improving teaching effectiveness.
[0005] To achieve the above objectives, this utility model provides the following technical solution: It includes a microfluidic driving device, a dynamic simulation module, and a teaching imaging system. The dynamic simulation module comprises, from top to bottom, an invasion layer, an intestinal simulation layer, and a vascular metastasis layer. The invasion layer contains lyophilized and stained colon cancer cells. The bottom of the invasion layer is composed of a capillary simulation plate simulating capillaries and having a micron-level pore structure. The intestinal simulation layer simulates intestinal tissue. The bottom of the intestinal simulation layer is composed of a common blood vessel simulation plate simulating ordinary blood vessels and having a micron-level pore structure. The vascular metastasis layer simulates the human body environment. The microfluidic driving device is connected to the input end within the vascular metastasis layer and simulates blood flow. The output end within the vascular metastasis layer is connected to a biosafety collection bottle. The teaching imaging system includes an excitation light source for exciting the stained markers, a long working distance objective lens for observing the dynamic simulation module, and a microscope camera for the dynamic simulation module.
[0006] By employing the above technical solution, during operation, pre-warmed culture medium is injected into the invasion layer, and after standing for a certain period of time, lyophilized and stained colon cancer cells are activated. Then, the colon cancer cells break through a capillary simulation plate simulating capillaries and enter the intestinal simulation layer simulating the intestine. Next, the colon cancer cells break through a common blood vessel simulation plate simulating ordinary blood vessels and enter the vascular metastasis layer simulating the human body environment. Finally, a microfluidic driving device simulates blood flow, allowing the colon cancer cells to metastasize within the human body environment. During this process, the stained markers are activated by an excitation light source, and real-time observation is performed using a long working distance objective lens and a microscope camera. The camera data can also be imported into a computer for analysis using specialized software. This simulation device has a simple structure, is easy to operate, and provides intuitive results, making it very suitable for experimental teaching and effectively improving teaching effectiveness. Furthermore, the microfluidic system formed by the micron-level well plate combined with the microfluidic driving device makes the simulation process more accurate and realistic. In addition, the output end within the vascular metastasis layer is connected to a biosafety collection bottle, making the operation safer.
[0007] The present invention is further configured such that: the ordinary blood vessel simulation plate is a standard plate simulating a complete blood vessel or a leakage plate simulating a leaking blood vessel.
[0008] By adopting the above technical solution, two different micron-sized porous structure plates simulate complete blood vessels and leaking blood vessels, making up for the lack of simulation of blood vessel leakage in traditional experiments and making the mechanism display richer.
[0009] The present invention is further configured such that: the vascular transfer layer has multiple branches, the branches being a liver simulation branch, a lung simulation branch, and a control branch, the liver simulation branch simulating liver tissue, the lung simulation branch simulating lung tissue, and the control branch being used for comparison with the liver simulation branch and the lung simulation branch.
[0010] By adopting the above technical solutions, the vascular transfer layer can simulate a single site or have simulated branches simulating different sites, making up for the lack of organ-oriented comparative simulation in traditional experiments and making the mechanism display richer.
[0011] The present invention is further configured such that the microfluidic drive device is a piezoelectric micropump with a single operating parameter.
[0012] By adopting the above technical solution, using a piezoelectric micropump as a microfluidic driving device, the number of high-precision devices can be reduced as much as possible while ensuring experimental accuracy, and the process does not require parameter adjustment, further reducing the difficulty of operation. Attached Figure Description
[0013] Figure 1 This is a perspective view of a specific embodiment of the present utility model;
[0014] Figure 2This is an exploded view of the dynamic simulation module. Detailed Implementation
[0015] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0016] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.
[0017] like Figure 1 — Figure 2 As shown, this utility model discloses a microfluidic dynamic simulation device for colorectal cancer metastasis suitable for experimental teaching, including a microfluidic driving device, a dynamic simulation module, and a teaching imaging system.
[0018] The dynamic simulation module comprises, from top to bottom, an invasion layer 1, an intestinal simulation layer 2, and a vascular metastasis layer 3.
[0019] The invasive layer 1 is used to place lyophilized and stained colon cancer cells (HCT116, CellTracker Red stained). The bottom of the invasive layer 1 consists of a capillary simulation plate 11 with a micron-sized pore structure, simulating capillaries. The capillary simulation plate 11 has a 20μm micron-sized pore structure and is made of Matrigel matrix gel.
[0020] The intestinal simulation layer 2 simulates intestinal tissue, containing collagen gel and epithelial cells. The bottom of the intestinal simulation layer 2 consists of a simulated blood vessel plate 21 with a micron-sized pore structure, mimicking ordinary blood vessels (arteries and veins). This plate can be a standard plate simulating a complete blood vessel or a leaking plate simulating a leaking vessel. The standard plate has an 8μm micron-sized pore structure, endothelial cells (HUVEC, CD31-FITC labeled), and Matrigel matrix. The leaking plate has a serrated crack structure (gradual width of 5-50μm) and is pre-filled with fibrin gel containing a PAI-1 inhibitor (with added phenol red pH indicator). When cancer cells penetrate the cracks, the area turns yellow (pH 6.0→5.5; cancer cells produce lactic acid from sugar alcohol, causing a local pH decrease), making leakage visually identifiable.
[0021] The vascular transfer layer 3 simulates the human body environment (37℃, pH 7.4, 5% CO2). Vascular transfer layer 3 can simulate a single site (liver or lung) or have multiple branches: a liver simulation branch 31, a lung simulation branch 32, and a control branch 33. Liver simulation branch 31 simulates liver tissue and contains hepatocytes, CCL20 chemokine, and DMEM culture medium containing 10% fetal bovine serum. Lung simulation branch 32 simulates lung tissue and contains lung epithelial cells, CXCL8 chemokine, and DMEM culture medium containing 10% fetal bovine serum. Control branch 33 is used for comparison with liver and lung simulation branches 31 and 32, and contains DMEM culture medium containing 10% fetal bovine serum.
[0022] The microfluidic drive device, a piezoelectric micropump 4, is connected to the input end within the vascular transfer layer 3 and simulates blood flow. The output end within the vascular transfer layer 3 is connected to a biosafety collection bottle (34 is the output pipeline). All fluid pipelines are fully enclosed, and the connection between the input and output ends uses a Luer-lock interface. To meet biosafety laboratory requirements, the biosafety collection bottle employs a sealing strategy of dual mechanical seals + liquid seal / negative pressure system + real-time pressure monitoring to prevent leakage of hazardous biological materials.
[0023] The teaching imaging system includes an excitation light source for exciting staining labels and a long working distance objective lens and a microscope camera for capturing dynamic simulation modules. The excitation light source is a dual-channel LED light source 5 (470nm excitation FITC, 555nm excitation CellTracker Red). The long working distance objective lens 6 is selected as 20×, NA0.4, and the USB microscope camera 7 is selected as 5 megapixels, 30fps. The long working distance objective lens 6 is used in conjunction with the microscope eyepiece, and can also be used with the USB microscope camera 7 for electronic imaging (the dual-channel LED light source 5, long working distance objective lens 6 and USB microscope camera 7 in the attached figure are for illustration only). The data is transmitted to the computer via a data cable.
[0024] The experimental procedure and data analysis are briefly described below.
[0025] During the experiment, 1 mL of pre-warmed culture medium (DMEM medium containing 37°C, 5% CO2, and 10% fetal bovine serum) was first injected into the invasion layer 1. The medium was left to stand and wait for the lyophilized colon cancer cells to activate. Simultaneously, a piezoelectric micropump 4 (fixed at 1 Hz / 10 μL / min) was started, and observation was performed using a microscope. Next, the red cancer cells breached the capillary simulation plate 11; then, the ordinary blood vessel simulation plate 21 changed color, and the penetration time was recorded. Finally, the colon cancer cells metastasized to various branches of the vascular metastasis layer 3, and the color changes were recorded. During this process, the image information could be imported into a computer for analysis (an open-source Python program generated in real-time: red-green overlay image (cancer cell-endothelial cell interaction), migration velocity curve (μm / min), and extravasation rate (%)).
[0026] When analyzing the obtained data, the software automatically compares the extravasation rate differences between the standard plate and the leakage plate; it exports red-green fluorescence overlay video, marks the hot spots of cancer cell-endothelial cell adhesion, compares the differences in metastasis of different branches, and discusses the influence of the tumor microenvironment.
[0027] In addition, for safety reasons, the contact surfaces of the dynamic simulation module are covered with a tearable PDMS film (replaced every semester), or an antibacterial coating can be used. During experiments, daily cleaning, efficient disinfection, contamination treatment, safety protection, proper storage, and regular testing are required to meet biosafety laboratory standards. All liquid contact parts are pre-sterilized and sealed (gamma irradiated).
[0028] In terms of the structure of the dynamic simulation module, the invasion layer 1, the intestinal simulation layer 2 and the vascular transfer layer 3 are all fixed by snap-fit and have openings at the top. The invasion layer 1 is provided with a cover plate 8 that closes the opening. The capillary simulation plate 11 at the bottom of the invasion layer 1 closes the opening of the intestinal simulation layer 2, and the ordinary vascular simulation plate 21 at the bottom of the intestinal simulation layer 2 closes the vascular transfer layer 3.
[0029] In summary, this simulation device clearly and intuitively demonstrates the entire process of colon cancer cell metastasis to organs by simulating the metastasis process of colon cancer cells. The combination of micron-level well plates and microfluidic drive devices makes the simulation process more accurate and realistic. The optional vascular simulation plates and comparable simulated branches of metastatic sites enrich the demonstration of the mechanism. The device is simple in structure, easy to use, and highly interactive, allowing students to master the techniques within a limited class time while effectively stimulating their interest in exploration. Biosafety is a primary concern, ensuring the safe conduct of the experiment.
Claims
1. A microfluidic dynamic simulation device for colon cancer metastasis suitable for experimental teaching, characterized in that: The device comprises a microfluidic driving device, a dynamic simulation module and a teaching imaging system, the dynamic simulation module comprises from top to bottom an invasion layer, an intestinal simulation layer and a vascular metastasis layer, the invasion layer is used for placing freeze-dried and dyed colon cancer cells, the bottom of the invasion layer is composed of a capillary simulation plate simulating capillary vessels and having a micrometer-level pore structure, the intestinal simulation layer simulates intestinal tissues, the bottom of the intestinal simulation layer is composed of a common vessel simulation plate simulating common vessels and having a micrometer-level pore structure, the vascular metastasis layer simulates human body environment, the microfluidic driving device is connected to an input end in the vascular metastasis layer and simulates blood flow, an output end in the vascular metastasis layer is connected to a biological safety collection bottle, the teaching imaging system comprises an excitation light source exciting dyed markers, a long working distance objective lens observing the dynamic simulation module and a microscopic camera shooting the dynamic simulation module. 2. The colon cancer metastasis microfluidic dynamic simulation device suitable for experimental teaching according to claim 1, characterized in that: The common vessel simulation plate is a standard plate simulating complete vessels or a leakage plate simulating leakage vessels. 3.The colon cancer metastasis microfluidic dynamic simulation device suitable for experimental teaching according to claim 1, characterized in that: The vascular metastasis layer is provided with multiple branches, the branches are respectively a liver simulation branch, a lung simulation branch and a control branch, the liver simulation branch simulates liver tissues, the lung simulation branch simulates lung tissues, and the control branch is used for comparison with the liver simulation branch and the lung simulation branch.
4. The colon cancer metastasis microfluidic dynamic simulation device suitable for experimental teaching according to claim 1, characterized in that: The microfluidic driving device is a piezoelectric micropump with a single working parameter.