TWM583456U - Microfluidic chip with bead retention structure and microfluidic channel structure - Google Patents

Abstract

This creation is a bead-loaded microfluidic structure, including: a microfluid sample inlet for microfluid samples to enter; a resistance increasing section connected to the microfluid sample inlet; a detection section having a first end and a second end Wherein the first end is connected to the resistance increasing section and is used to test or process the microfluidic sample, and the beads are arranged in the detection section; and the bead system retention structure coupled to the second end of the detection section . The particle size of the beads is larger than the pore diameter of the bead structure, so that the bead system remains in the detection section.

Description

Micro-flow channel wafer with bead system retention structure and micro-flow channel structure

This creation is about a microfluidic wafer and microfluidic structure that increases the capture rate of biological material, especially a microfluidic wafer and microfluidic structure with a bead system retention structure, which keeps the beads stationary at the detection Section.

The high mortality rate caused by cancer in recent years has been a serious threat to human life. Studies have found that in the early stages of tumor development, most of them are organ-limited diseases, but in the later stages, they are almost always transmitted to the remote organs through the bloodstream to form metastases. Such remote metastases are often the main cause of patient death. Cells that detach from the primary site of the tumor and further enter the blood circulation system are called circulating tumor cells (CTC). CTC is considered to be one of the important causes of tumor metastasis. The number of particles and surface molecules The expression of markers has an important index effect on the post-healing judgment and efficacy evaluation of tumor patients.

Depending on the tumor itself and the response to chemotherapy and drug treatment, the number of CTCs will change dynamically in real time. Therefore, this feature can be used for early diagnosis in vitro, rapid evaluation of drugs, and reference for personalized treatment. However, in the blood of cancer patients, CTC is a rare cell, and there is only one CTC every 10 ⁹ blood cells, making it difficult to detect and isolate CTC technology. Therefore, a centralized collection method must be used to effectively detect and isolate CTCs.

One example of the current centralized collection method is the use of cell surface biomarkers with high specificity and sensitivity to CTC, such as epithelial cell adhesion molecules (EpCAM). Nagrath et al. (Nature 2007, 450: 1235-9) developed anti-EpCAM antibody-coated microfluidic wafers for CTC detection and collection. However, the shortcomings of the above techniques are low detection rate and low collection purity. This is due to the non-specific binding of blood cells to anti-EpCAM antibodies and the fluid design structure.

Despite advances in the technology for detecting and isolating CTCs, more specific and effective methods are needed to detect, purify, and release CTCs and other biological substances for further cultivation and characterization.

The reason for this is that the applicant, in view of the lack of known technology, has carefully experimented and researched, and has persisted in a spirit of perseverance, and finally created the case "microchannel chip and microchannel structure with bead system retention structure" In order to improve the lack of the conventional techniques mentioned above.

This creation is a new type of microfluidic system, which includes a microfluidic wafer and beads located in the microfluidic wafer and capable of grasping circulating tumor cells to separate circulating tumor cells from blood cells. The created microfluidic structure and microfluidic wafer especially include a resistance increasing section and a bead system retention structure, so that the bead system is left in the detection main area, which is convenient for users to observe the microfluidic structure and the microfluidic chip. Adsorption of beads.

The principle of the created microfluidic system is to use the table of circulating tumor cells The characteristics of the surface antigen and the antibodies implanted on the surface of the bead are used for grasping. The bead structure results in the largest contact area per unit volume, followed by the fluid resistance and the curved structure of the microchannel structure, which cause disturbances and cause circulating tumors. Cells rotate or roll and increase the chance of contact with the beads to enhance the grasping effect, and the special design of the microchannel structure reduces the non-specific binding of blood cells to anti-EpCAM antibodies.

One aspect of this creation is to provide a micro-flow channel wafer carrying a bead having a particle size, including: a substrate; a body having a first surface and a second surface, wherein the second surface is closely adhered Covering the substrate; and a microfluidic channel structure embedded in the second surface, so that the microfluidic channel structure forms a microfluidic channel between the body and the substrate, wherein the microfluidic channel structure includes: a blood The sample inlet extends from the first surface to the second surface and has a diameter for a blood sample to enter; an expansion section connected to the blood sample inlet and has a first width; a resistance increasing section, Connected to the expansion section, having a second width; a detection section connected to the resistance increasing section, and the bead is arranged in the detection section; and a slow-flow section, connected to the detection section The measurement section is connected with a first depth, wherein the particle diameter is larger than the first depth to prevent the beads from entering the slow-flow section, and the second width is smaller than the first width and the diameter to prevent the The blood sample flows back to the expanded section, which makes the bead system In the detection zone.

Another aspect of this creation is to provide a microchannel structure carrying a bead with a particle size, including a structural body, for allowing a microfluid sample to flow through the microchannel structure for inspection or processing. The structure body includes: a microfluid sample inlet with a first aperture for the microfluid sample to enter; a resistance increasing zone Segment, which is connected to the microfluid sample inlet and has a second aperture; a detection segment having a first end and a second end, wherein the first end is connected to the resistance increasing segment and is used for Inspecting or processing the microfluidic sample, wherein the bead is disposed in the detection section; and a bead system retention structure coupled to the second end for retaining the bead in the detection section .

In order to make the above and other objects, features, and advantages of this creation more comprehensible, the preferred embodiments are described below in conjunction with the accompanying drawings for a detailed description.

10‧‧‧Micro channel chip

100‧‧‧ substrate

200‧‧‧ Ontology

210‧‧‧first surface

220‧‧‧ second surface

300‧‧‧Micro channel structure

310‧‧‧ Blood sample entrance

320‧‧‧ Expansion Section

321‧‧‧ the first end

322‧‧‧ second end

330‧‧‧Resistance section

340‧‧‧ Detection section

341‧‧‧ the first end

342‧‧‧Detect main area

343‧‧‧second end

344‧‧‧Bead system retention structure

350‧‧‧ Slow stream section

360‧‧‧ blood sample export

40‧‧‧ beads

50‧‧‧Micro channel structure

500‧‧‧ structure ontology

510‧‧‧Microfluid sample inlet

520‧‧‧Increase resistance section

530‧‧‧ Detection section

532‧‧‧bead system retention structure

540‧‧‧ Slow section

550‧‧‧Microfluidic sample outlet

60‧‧‧ Beads

Figure 1 is a schematic plan view of the creative microfluidic wafer; Figure 2 is a schematic diagram of the vertical section of AA 'in Figure 1; Figure 3 (A) is the detection of the creative microfluidic wafer. Schematic diagram of setting up beads in section; Figure 3 (B) is a schematic diagram of setting up beads in the detection section of another embodiment of the creative microfluidic chip; Figure 4 is a diagram of the second end of the detection section The effect of different widths on the cell recovery rate; Figure 5 is a schematic diagram of the depth of the slow-flow section in the microchannel wafer smaller than the particle size of the beads; Figure 6 is a schematic diagram of the microchannel structure in another embodiment of the creation ; Figure 7 is a result chart of the recovery efficiency and detection limit of the microfluid-free system; Figure 8 is a result chart of the recovery efficiency and detection limit of the creative microfluidic wafer.

The following describes the embodiments of the "microchannel wafer and microchannel structure with a bead system retention structure" in this case. Please refer to the drawings, but the actual configuration and the method adopted do not necessarily conform to the description. Those skilled in the art can make various changes and modifications without departing from the actual spirit and scope of this case.

The created micro-fluidic wafer is loaded with beads. The beads are particularly large beads with a particle size of 100 to 200 μm. The bead surface coating includes (1) the release or removal of non-specific blood cells and other blood components ( (E.g., a protein) releasable composition; (2) a biologically active composition that captures a biological substance; or (3) a linking molecule linked to a releasable composition and a biologically active composition. When the microfluidic sample flows through the bead, the bead can capture the biological material in the microfluidic sample that can react with the reactive material on the surface of the bead, and can release the captured biological material for further research and detection. The material of the beads is transparent plastic or transparent resin. The microfluidic sample may be a body fluid or a bacterial fluid, and the body fluid includes blood, cerebrospinal fluid, various digestive fluids, semen, saliva, sweat, urine, vaginal fluid, or a solution containing biological substances. Biological substances include CTC, CTC circulating stem cells (such as tumor stem cells, liver stem cells, and bone marrow stem cells), fetal cells, bacteria, viruses, epithelial cells, endothelial cells, or other biological substances. Therefore, depending on the object to be grasped, the substance coated on the bead surface is also different.

An example of this creation is the separation of circulating tumor cells from the blood. The microfluidic chip has a plurality of transparent beads inside. When the beads capture the circulating tumor cells, they will be separated from the blood and positioned in the detection section. The remaining normal blood cells will flow out of the outlet and flow into the waste liquid storage tank. In order to capture and isolate circulating tumor cells in the blood, the substance coated on the surface of the beads is preferably an antibody against epithelial cell adhesion molecules (Epithelial Cell Adhesion Molecule, EpCAM).

Please refer to Figs. 1, 2, 3 (A) and 3 (B), which are a schematic plan view of the microfluidic wafer and a vertical cross-sectional view extending along A-A 'for the creation. The created microfluidic wafer 10 includes a substrate 100, a body 200 and a microfluidic structure 300. The body 200 has a first surface 210 and a second surface 220 opposite to the first surface 210. The microchannel structure 300 is embedded in the second surface 220 of the body 200, and the second surface 220 is closely covered on the substrate 100. The microfluidic channel structure 300 is formed between the body 200 and the substrate 100.

The created microfluidic channel structure 300 includes a blood sample inlet 310, an expansion section 320, a resistance increasing section 330, a detection section 340, a slow flow section 350, and a blood sample outlet 360 in order from the entrance to the exit.

The original blood sample inlet 310 extends from the first surface 210 to the second surface 220 of the body 200 for blood samples to enter the flow channel. The blood sample inlet 310 may be a circular hole or a polygonal hole, preferably a circular hole. The diameter of the blood sample inlet 310 of this creation is between 0.8 and 1.2 mm, and it can accommodate a syringe with an 18 to 21 G needle (about 0.7 to 0.9 mm).

The creative expansion section 320 has a first end 321 and a second end 322. The first end 321 is connected to the blood sample inlet 310, and the second end 322 is connected to the resistance increasing section 330. The aperture of the expansion section 320 may be circular or polygonal, preferably square. The creative extension section 320 has a width between 0.8 and 1.5 mm and a depth of 1 mm.

One end of the creative resistance increasing section 330 is connected to the second end 322 of the expansion section 320, and the other end is connected to the detection section 340. The hole diameter of the resistance increasing section 330 may be circular or polygonal, preferably square, and the width of the resistance increasing section 330 will be smaller than the width of the expansion section 320 and the blood sample inlet 310, and larger than the particle diameter of the beads, resulting in When the beads are passed, the fluid resistance can be enhanced, and the function of preventing the liquid from flowing back due to the needle insertion and withdrawal can be prevented. The width of the created resistance increasing section 330 is between 250 μm and the depth is 1 mm. Since the width of the expansion section 320 is greater than the width of the resistance increasing section 330, the structure of the second end 322 of the expansion section 320 can be gradually reduced from the width of the expansion section 320 to the width of the resistance increasing section 330, that is, the second The width of the end 322 is gradually reduced from 0.8 to 1.5 mm to 250 μm.

The creation detection section 340 includes a first end 341, a detection main area 342, and a second end 343. The first end 341 is connected to the resistance increasing section 330, and the second end 343 is connected to the slow flow section 350. The main measurement region 342 is located between the first end 341 and the second end 343, and a bead 40 (as shown in Figs. 3 (A) and 3 (B)) capable of adsorbing circulating tumor cells in the blood is provided. The aperture of the detection section 340 may be circular or polygonal, preferably square. In the embodiment of the present invention, the aperture of the detection section 340 is square. In order for the bead 40 to be disposed in the detection main area 342 in a single layer, the detection section 340 is a region that limits the depth of the flow channel, and the depth of the detection section 340 is the particle diameter of the bead 40 plus 20 to 50 μm. Therefore, the depth of the detection section 340 is between 120 and 250 μm. The width of the first end 341 and the detection main area 342 of the detection section 340 is sufficient for the bead 40 to pass through, so the width is between 250 μm and 1.5 mm. The width of the first end 341 of the detection section 340 may be the same as the width of the resistance increasing section 330, or may gradually increase from the width of the resistance increasing section 330 to the first The width of one end 341. The detection main area 342 can hold approximately 20 to 30 beads 40. The width of the second end 343 of the detection section 340 affects the arrangement of the beads 40 at the second end 343, and the arrangement of the beads 40 at the second end 343 of the detection section 340 affects the flow direction of the liquid. According to the experimental results in Figure 4, when the bead is 200 μm and the width of the second end 343 of the detection section 340 is 250 μm (that is, it can only accommodate one bead 40), the cell recovery rate is the best. And as the width of the second end 343 increases, the cell recovery rate decreases. Therefore, the width of the second end 343 of the detection section 340 may be a width that can accommodate one bead. The width of the second end 343 of the detection section 340 of the present creation is between 150 and 250 μm. Since the width of the detection main region 342 is greater than the width of the second end 343, the structure of the second end 343 can be gradually changed from the width of the detection main region 342 (as shown in FIG. 3 (A)) or in a stepwise manner (As shown in FIG. 3 (B)) is reduced to the width of the second end 343.

In order to make the bead 40 stay in the detection main area 342 and not move with the flow of the liquid, the microchannel structure 300 of the present creation includes a bead system retaining structure 344. The bead system retention structure 344 is coupled to the second end 343 of the detection section 340, and the pore diameter of the bead system retention structure 344 is smaller than the particle diameter of the bead 40, so that the bead 40 cannot pass through the bead system retention structure 344 and enter the buffer. The flow section 350 causes the beads 40 to remain in the detection main area 342. In addition, the resistance-increasing section 330 can prevent the blood sample from flowing back from the detection section 340 to the expansion section 320 due to the needle insertion, so that the bead 40 will not move with the needle insertion, and The bead 40 is stably stayed in the detection main area 342 to facilitate observation of the state of the bead 40 adsorbing the biological substance. The bead system retention structure 344 of this creation may be the second end 343 of the detection section 340, so in this case, the pore diameter of the second end 343 of the detection section 340 is smaller than the particle diameter of the bead 40 (such as the third (B).

In another embodiment, in order to keep the bead 40 in the detection main area 342, the bead system retention structure 344 may also be a slow flow section 350. Therefore, in this case, the depth of the slow flow section 350 is less than The particle diameter of the beads 40 (as shown in FIG. 5) prevents the beads 40 from entering the slow-flow section 350. Therefore, the bead system retention structure 344 is coupled to the second end 343 of the detection section 340, which means that the bead system retention structure 344 is the second end 343 of the detection section 340, or is connected to the second end of the detection section 340. End 343.

One end of the slow-flow section 350 of this creation is connected to the second end 343 of the detection section 340 and the other end is connected to the blood sample outlet 360. The aperture of the slow-flow section 350 may be circular or polygonal, preferably square. In the embodiment of the present invention, the width of the slow-flow section 350 is between 150 and 250 μm, and the depth is between 50 and 100 μm.

One end of the creative blood sample outlet 360 is connected to the slow-flow section 350, and the other end extends from the second surface 220 to the first surface 210 of the body 200. Blood cells not captured by the bead 40 will flow to the waste liquid recovery area (not shown) through the blood sample outlet 360. The blood sample outlet 360 may be a round hole or a square hole, preferably a round hole, and its diameter is between 0.8 and 1.2 mm.

The following table 1 is a preferred embodiment of the particle diameter of the bead 40 and the pore diameter of each section in the microchannel structure 300.

The material of the substrate 100 can be acrylic (polymethylmethacrylate, PMMA), polyethylene terephthalate (PET), polycarbonate (PC), or polydimethylsiloxane ( polydimethylsilicon (PDMS), silicone, rubber, plastic or glass. The material of the body 200 may be polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC), polydimethylsilicon (PDMS) ), Silicone, rubber, or plastic. When selecting materials for the substrate 100 and the body 200, it is necessary to consider the substrate Material characteristics between 100 and body 200. In the embodiment of the present invention, the substrate 100 is glass, and the body 200 is polydimethylsiloxane.

The present invention further provides another embodiment of the microfluidic channel structure 50, as shown in FIG. 6. The microchannel structure 50 contains beads 60 and has a structure body 500. The structure body 500 includes a microfluid sample inlet 510, a resistance increasing section 520, a detection section 530, a slow flow section 540, and The fluid sample outlet 550, wherein the bead 60 is located in the detection section 530. A bead retention structure 532 is coupled between the detection section 530 and the slow flow section 540 to retain the bead 60 in the detection section 530. After the microfluidic sample enters from the microfluidic sample inlet 510, it can directly enter the detection section 530 through the resistance increasing section 520, and the beads 60 in the detection section 530 capture the biological material in the microfluidic sample to After the microfluidic sample is inspected or processed, it enters the slow flow section 540, and finally flows out of the microfluidic channel structure 50 from the microfluidic sample outlet 550.

The method of preparing the micro-fluidic wafer of this creation is to first use a 3D printer to print a master mold. The master mold is a light-curing resin, which is rinsed with 95% alcohol, and then cured by UV light for 2 minutes. Bake for 10 minutes. The food-grade material PDMS liquid was poured into the master mold in proportion to the curing process for 50 minutes and 80 degrees, and then bonded to the glass substrate with an oxygen plasma machine.

Experimental example

Study on the Circulating Tumor Cell Beads in Cultured Circulating Tumor Cells

1. Recovery efficiency and detection limit of large beads (200 microns in diameter) in microfluid-free systems

Put 10, 1,000, and 100,000 circulating tumor cells and beads and 1 mL of physiological saline buffer solution (simulating blood environment) into a centrifuge tube, and fully circulate the tumor cells and beads in physiological saline buffer solution. After mixing, observe the grasping efficiency of the beads. According to Figure 7, the experimental results show that only 1.5% of the cells (about 1500) were captured by the beads in the experimental group with 100,000 circulating tumor cells, but the experiments with 10 and 1000 circulating tumor cells In the group, the beads did not capture any circulating tumor cells, which means that the blood environment of less than 1000 circulating tumor cells, the beads could not capture any circulating tumor cells.

2.Recycling efficiency and detection limit of large beads (200 microns in diameter) in the microchannel wafer

10, 50, 100, 500, and 1000 circulating tumor cells were mixed with 1 mL of physiological saline buffer solution, and the mixed liquid sample was passed through the beads in the microchannel wafer of this creation, and the beads were observed Body grabbing efficiency. According to Figure 8, the experimental results show that using the microfluidic wafer of this creation, a liquid sample containing more than 50 circulating tumor cells can be grasped, compared to the results obtained without a microfluidic system (requiring 100,000 cells to (Captured, as shown in Figure 8), the detection limit is significantly reduced by 2000 times, and the recycling efficiency of the micro-channel wafer using this creation is on average higher than 5%, which is about 3 times higher than that of the microfluid-free system. .

When an average of about 50 circulating tumor cells in the blood of a human body per 10 mL, it means that the person has a high risk of cancer. Therefore, this experiment proves that only large beads (200 micrometers in diameter) cannot distinguish the risk of human cancer. However, the large beads combined with the microchannel chip created by this method can effectively and accurately grasp the blood. Circulating tumor cells can more quickly determine whether they have cancer.

After the blood samples of cancer patients were actually stained, the microfluidic wafer separation results (see the attached drawings 1 to 3) of the original creation, in which green is circulating tumor cells and red is white blood cells. Figure 1 shows that the circulating tumor cells (green dots) are captured on the transparent beads, Figure 2 is that the white blood cells (red dots) are incorrectly captured, and Figure 3 is obtained by combining Figures 1 and 2 Location of all captured cells. This experiment demonstrates the results exhibited by patients with stage II cancer in the microfluidic chip of this creation. The results show that about 13 circulating tumor cells were captured, and only 3 white blood cells were captured (because of 1mL of blood in the human body) The number of white blood cells is between about 10 ⁶ and 10 ^7. According to strict definition, it is estimated by 10 ⁶ white blood cells, that is, only 3 white blood cells are captured by one million white blood cells, which is far lower than the current FDA proof. Cellsearch instrument's catch rate (10 ⁶ white blood cells caught about 3000 ~ 4000)). In addition, the results of this experiment took only 30 minutes from the blood sample acquisition to the imaging results display, compared with the previous pre-processing and circulating tumor cell separation to read the imaging results in 6-9 hours, which is much shorter in time. Therefore, using the microfluidic chip of this creation can effectively capture a small amount of circulating tumor cells in the blood, has a very low error rate, and the result can be obtained in only 30 minutes, so the microfluidic chip of this creation It can be used as a quick-screen biochip for preliminary detection of cancer.

Other embodiments

1. A microchannel structure carrying a bead having a particle size, comprising a structure body for passing a microfluid sample through the microchannel structure for inspection or processing, wherein the structure body includes : A microfluidic sample inlet with a first An aperture for the microfluidic sample to enter; a resistance increasing section connected to the microfluidic sample inlet and having a second aperture; a detection section having a first end and a second end, wherein the The first end is connected to the resistance increasing section and is used for testing or processing the microfluidic sample, wherein the bead is disposed in the detection section; and a bead system retention structure is coupled to the second end, Used to tether the bead in the detection section.

2. The microchannel wafer according to embodiment 1, wherein the particle diameter is 100 to 200 μm, the diameter is 0.8 to 1.2 mm, the first width is 0.8 to 1.5 mm, the second width is 250 μm, and the The first depth is 50 to 100 μm.

3. The micro-channel wafer according to embodiment 2, wherein when the particle diameter is 100 μm, the first depth is 50 μm, and when the particle diameter is 200 μm, the first depth is 100 μm.

4. The micro-flow channel chip according to embodiment 2, wherein the depth of the expansion section and the resistance increasing section is 1 mm, the width of the detection section is 250 μm to 1.5 mm, and the depth of the detection section 20 to 50 μm is added to the particle diameter, so that the beads are retained in the detection section as a single layer, and the detection section accommodates 20 to 30 of the beads.

5. The microfluidic chip according to embodiment 1, wherein the expansion section includes a first end and a second end, the first end is connected to the blood sample inlet, and the second end is connected to the resistance increasing region The segments are connected, and the width of the second end is gradually reduced from the first width to the second width.

6. The micro-channel wafer according to embodiment 1, wherein the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate (polyethylene terephthalate, PET), polycarbonate (PC), polydimethylsilicon (PDMS), silicone, rubber, plastic, or glass, and the material of the body is polymethylmethacrylate (PMMA) ), Polyethylene terephthalate (PET), polycarbonate (PC), polydimethylsilicon (PDMS), silicone, rubber or plastic.

7. The micro-channel wafer according to embodiment 1, wherein the first surface is disposed opposite to the second surface.

8. A microchannel structure carrying a bead having a particle size, comprising a structure body for passing a microfluid sample through the microchannel structure for inspection or processing, wherein the structure body includes : A microfluidic sample inlet having a first aperture for the microfluidic sample to enter; a resistance increasing section connected to the microfluidic sample inlet and having a second aperture; a detection section having a first aperture One end and a second end, wherein the first end is connected to the resistance increasing section and is used for testing or processing the microfluidic sample, wherein the bead is arranged in the detection section; and a bead system is left The structure is coupled to the second end for retaining the bead in the detection section.

9. The microfluidic channel structure according to embodiment 8, wherein the second aperture is smaller than the first aperture to prevent the microfluidic sample from flowing back to the microfluidic sample inlet.

10. The micro-flow channel structure according to embodiment 8, wherein the structure body further comprises a slow-flow section connected to the second end of the detection section, and the slow-flow section has a third aperture, And the second end of the detection section has a fourth aperture.

11. The microchannel structure according to embodiment 10, wherein the bead system remains The structure is the second end of the detection section or the slow-flow section.

12. The microchannel structure according to embodiment 11, wherein when the bead system retention structure is the second end of the detection section, the third pore size is smaller than the particle diameter, and when the bead system retention structure is In the slow-flow section, the fourth pore diameter is smaller than the particle diameter to retain the beads in the detection section.

13. The micro-channel structure according to embodiment 12, wherein the first aperture is a circular hole having a diameter, and wherein: the particle diameter is 100 to 200 μm; the diameter is 0.8 to 1.2 mm; the second aperture The width of the fourth aperture is 250 μm and the depth is 1 mm; the width of the third aperture is 150 to 250 μm; the depth is the particle size plus 20 to 50 μm; and the width of the fourth aperture is 150 to 250 μm and the depth is 50 to 100 μm.

14. The microchannel structure according to embodiment 12, wherein a width of the third aperture is the same as a width of the fourth aperture.

15. The microchannel structure according to embodiment 8, wherein the microfluidic sample is a body fluid or a bacterial fluid.

In summary, the new model can indeed use a novel concept to match the microfluidic chip and large beads created by this invention, which can effectively capture a small amount of circulating tumor cells in the blood and reduce the rate of error. To determine the occurrence of cancer at an early stage. In addition, the microfluidic chip created by the creation of the resistance-increasing section and the bead system retention structure allows the beads to stay in the detection section statically, so that the user can observe the status of the beads adsorbing biological substances. Furthermore, the microfluidic chip only requires 20 to 30 large beads, which can greatly reduce the manufacturing cost. Therefore, those who are familiar with this technique can be modified by various techniques, but they are not inferior to those who want to protect the scope of patent application.

Claims (15)

A micro-flow channel wafer carrying a bead having a particle size, comprising: a substrate; a body having a first surface and a second surface, wherein the second surface is closely covered on the substrate; and A microchannel structure is embedded on the second surface, so that the microchannel structure forms a microchannel between the body and the substrate, wherein the microchannel structure includes a blood sample inlet from the first The surface extends to the second surface and has a diameter for a blood sample to enter; an expansion section connected to the blood sample inlet having a first width; a resistance increasing section connected to the expansion section, Has a second width; a detection section connected to the resistance increasing section and the bead is arranged in the detection section; and a slow flow section connected to the detection section and has a A first depth, wherein the particle diameter is greater than the first depth to prevent the beads from entering the slow-flow section, and the second width is smaller than the first width and the diameter to prevent the blood sample from flowing back to the expansion zone Segment, thereby leaving the bead system in the detection segment. The micro-channel wafer according to item 1 of the patent application range, wherein the particle diameter is 100 to 200 μm, the diameter is 0.8 to 1.2 mm, the first width is 0.8 to 1.5 mm, and the second width is 250 μm, and The first depth is 50 to 100 μm. The micro-channel wafer according to item 2 of the patent application, wherein when the particle diameter is 100 μm, the first depth is 50 μm, and when the particle diameter is 200 μm, the first depth is 100 μm. According to the micro channel chip described in the second patent application range, wherein the depth of the expansion section and the resistance increasing section is 1 mm, the width of the detection section is 250 μm to 1.5 mm, The depth is the particle size plus 20 to 50 μm, so that the beads are tied in the detection section in a single layer, and the detection section accommodates 20 to 30 of the beads. The microfluidic chip as described in the first patent application range, wherein the expansion section includes a first end and a second end, the first end is connected to the blood sample inlet, and the second end is connected to the resistance increase The segments are connected, and the width of the second end is gradually reduced from the first width to the second width. The micro-fluidic wafer as described in item 1 of the patent application scope, wherein the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), and polycarbonate (PC), polydimethylsilicon (PDMS), silicone, rubber, plastic or glass, and the material of the body is polymethylmethacrylate (PMMA), polyethylene terephthalate (polyethylene terephthalate (PET), polycarbonate (PC), polydimethylsilicon (PDMS), silicone, rubber or plastic. The micro-flow channel wafer according to item 1 of the patent application scope, wherein the first surface is disposed opposite to the second surface. A microchannel structure carrying a bead with a particle size includes a structure body for passing a microfluid sample through the microchannel structure to be inspected or processed. The structure body includes: a The microfluid sample inlet has a first aperture for the microfluid sample to enter; a resistance increasing section connected to the microfluid sample inlet and has a second aperture; a detection section having a first end And a second end, wherein the first end is connected to the resistance increasing section, and is used for testing or processing the microfluid sample, wherein the bead is arranged in the detection section; and a bead system retention structure, Coupled to the second end for retaining the bead in the detection section. The microfluidic channel structure described in item 8 of the patent application scope, wherein the second aperture is smaller than the first aperture to prevent the microfluidic sample from flowing back to the microfluidic sample inlet. The micro-channel structure according to item 8 of the patent application scope, wherein the structure body further includes a slow-flow section connected to the second end of the detection section, and the slow-flow section has a third aperture And the second end of the detection section has a fourth aperture. The micro-channel structure according to item 10 of the patent application scope, wherein the bead system retention structure is the second end of the detection section or the slow-flow section. The micro-channel structure according to item 11 of the scope of patent application, wherein when the bead system retention structure is the second end of the detection section, the third pore size is smaller than the particle diameter, and when the bead system retention structure is For the slow-flow section, the fourth pore diameter is smaller than the particle diameter to retain the beads in the detection section. The micro-channel structure according to item 12 of the application, wherein the first pore diameter is a circular hole with a diameter, and wherein: the particle diameter is 100 to 200 μm; the diameter is 0.8 to 1.2 mm; and the second The width of the aperture is 250 μm and the depth is 1 mm; the width of the third aperture is 150 to 250 μm, the depth is the particle size plus 20 to 50 μm; and the width of the fourth aperture is 150 to 250 μm and the depth is 50 to 100 μm. The micro-channel structure according to item 12 of the application, wherein the width of the third aperture is the same as the width of the fourth aperture. The microfluidic channel structure described in item 8 of the scope of the patent application, wherein the microfluidic sample is a body fluid or a bacterial fluid.