Tumor cell counting device based on micro-fluidic chip and use method

Abstract

The invention discloses a tumor cell counting device based on a micro-fluidic chip and a use method. The tumor cell counting device comprises the micro-fluidic chip, an external pump thrust device, a mixing part, a micro-fluidic part, a confluence part and an external collection device. The external pump thrust device is used for controlling the reaction liquid to flow among the functional areas of the micro-fluidic chip; the mixing part is arranged in the micro-fluidic chip; the micro-flow part is arranged on one side of the mixing part and is communicated with the mixing part; the confluence part is arranged on one side, far away from the mixing part, of the micro-flow part and is communicated with the micro-flow part. Fluid is input into the first inlet and the second inlet of the micro-fluidic chip through an external pump thrust device, the fluid enters the mixing part and is mixed by utilizing potential energy of the fluid, cells in the mixed fluid mixed solution are marked and enter the micro-flow part, and the micro-flow part captures target cells in the fluid and finally flows into the confluence part.

Description

Technical Field

[0001] The present invention relates to the technical field of microfluidic chips, and in particular to a tumor cell counting device based on a microfluidic chip and a use method thereof. Background Art

[0002] The MCF-7 cell line grows adherently and exhibits epithelial cell morphology under a microscope. MCF-7 cells grow relatively slowly, usually requiring 6-12 days to reach a growth density of 80%. MCF-7 cells are estrogen receptor (ER) and progesterone receptor (PR) positive and belong to the luminal A type molecular subtype. They grow hormone-dependent, but a small amount of hormone in the culture medium can meet their growth needs. MCF-7 cells retain multiple characteristics of differentiated mammary epithelium, such as the ability to process estradiol through cytoplasmic estrogen receptors and form bulge structures, and express the WNT7B oncogene. MCF-7 cells have extensive aneuploidy and heterogeneity in gene expression, and different subclones differ in gene expression, receptor expression, and signaling pathways. Summary of the invention

[0003] The present invention overcomes the deficiencies of the prior art and provides a tumor cell counting device based on a microfluidic chip and a use method thereof.

[0004] To achieve the above object, the technical solution adopted by the present invention is: a tumor cell counting device based on a microfluidic chip and a method of use, comprising:

[0005] Microfluidic chips,

[0006] An external pump thrust device is used to control the flow of the reaction liquid between the functional areas of the microfluidic chip;

[0007] A mixing unit, disposed in the microfluidic chip;

[0008] A first inlet, disposed on the upper end surface of the microfluidic chip and connected to the mixing portion;

[0009] A second inlet is provided on the upper end surface of the microfluidic chip and is connected to the mixing part;

[0010] A microfluidic section, disposed on one side of the mixing section and communicated with the mixing section;

[0011] a confluence portion, disposed on a side of the microfluidic portion away from the mixing portion and communicated with the microfluidic portion; and

[0012] An external collection device, arranged above the microfluidic chip, for collecting flow and quantity of the microfluidic chip;

[0013] In a preferred embodiment of the present invention, the mixing section comprises:

[0014] A mixing chamber is disposed in the microfluidic chip and has a circular cross section;

[0015] A discharge port, which is arranged at the middle of the lower side of the first mixing chamber, is communicated with the first mixing chamber, and has a diameter smaller than the cross-sectional diameter of the first mixing chamber;

[0016] A first feed channel is tangentially arranged along the outer wall of the first mixing chamber and is connected to the first mixing chamber and the first inlet, respectively; and

[0017] A second feed channel is arranged tangentially along the outer wall of the first mixing chamber, in a direction perpendicular to the first feed channel, and is connected to the first mixing chamber and the second inlet, respectively, and one end of the second feed channel is arranged close to the first feed channel;

[0018] In a preferred embodiment of the present invention, the microfluidic section comprises:

[0019] A plurality of microfluidic channels are arranged side by side in the microfluidic chip, each of which has a diameter of 50.0-60.0 μm. A plurality of branch channels are arranged at intervals in the microfluidic channels, and the single-side diameter of the branch channels is the same as the diameter of the microfluidic channel.

[0020] A plurality of capture claws are respectively arranged in the plurality of forks, and the side surfaces thereof are arranged in a streamlined shape. A plurality of capture claws are respectively arranged with a notch on the side facing the discharge port, and the notch has a diameter of 20-30 μm.

[0021] A plurality of flow-dividing channels are respectively arranged between the discharge port and the plurality of micro-fluidic channels, and are respectively connected with the discharge port and the plurality of micro-fluidic channels. The diameters of the plurality of flow-dividing channels are respectively the same as the diameters of the plurality of micro-fluidic channels.

[0022] In a preferred embodiment of the present invention, the microfluidic section comprises:

[0023] A plurality of screening ports are respectively arranged in the plurality of capturing claws, respectively connected to the plurality of recesses, and penetrate the plurality of capturing claws, and the diameter of the plurality of screening ports is 12-18 μm;

[0024] In a preferred embodiment of the present invention, the confluence portion comprises:

[0025] A plurality of converging channels are respectively arranged on a side of the plurality of microfluidic channels away from the discharge port and are respectively connected to the plurality of microfluidic channels;

[0026] A flow collecting port, which is arranged on the upper end surface of the microfluidic chip and is connected with a plurality of the flow collecting channels;

[0027] In a preferred embodiment of the present invention, the recesses of the plurality of capture claws are respectively coated with ER antibodies;

[0028] In a preferred embodiment of the present invention, the external acquisition device includes:

[0029] Data processors;

[0030] a flow sensor, disposed at the discharge port and electrically connected to the data processor; and

[0031] An image collector is disposed above the microfluidic channels and connected to a data processor;

[0032] In a preferred embodiment of the present invention, it includes:

[0033] A collecting tank is arranged on the upper end surface of the microfluidic chip and is connected to the collecting port;

[0034] In a preferred embodiment of the present invention, the microfluidic chip material is PDMS;

[0035] In a preferred embodiment of the present invention, a method for using the microfluidic chip-based tumor cell counting device comprises:

[0036] Step 1: First, a cell flow containing MCF-cells and a fluorescent agent are respectively input into the first inlet and the second inlet through an external inference device, and mixed in the microfluidic chip to obtain a mixed solution;

[0037] Step 2: The mixed solution is split and introduced into microfluidic channels with different narrow and wide alternations, so that the cells are focused on the center line of the fluid and captured by the capture claws;

[0038] Step 3: Collect images of the microfluidic channel and count based on the images;

[0039] Step 4: Obtain the actual tumor content based on the capture rate and counts;

[0040] The present invention solves the defects existing in the background technology and has the following beneficial effects:

[0041] (1) The present invention includes a microfluidic chip, an external pump thrust device, a mixing part, a first inlet, a second inlet, a microfluidic part, a confluence part, and an external collection device. Fluids are respectively input into the first inlet and the second inlet of the microfluidic chip through the external pump thrust device, and the fluids enter the mixing part and are mixed using their own potential energy. The cells in the mixed fluid mixture are labeled and enter the microfluidic part, and the microfluidic part captures the target cells in the fluid, and finally flows into the confluence part. Through the simple channel and chamber design, MCF-7 cells are labeled and captured. Through the setting of the flow collector, the flow of several fluids entering the microfluidic channel can be measured in real time, and the number of captured cells can be observed through the external collection device. According to the capture rate and count, the real tumor content can be obtained. It is easy to use and has strong practicality. The number and concentration of cells can be obtained in a short time, providing reliable basic values ​​for subsequent medical research.

[0042] (2) The present invention includes a mixing chamber, a discharge port, a first feed channel, and a second feed channel. The fluid enters the first mixing chamber through the first feed channel and the second feed channel respectively, and the right-angle impact principle is used to make the fluid fully mixed by itself, and then the fluid rotates downward to enter the discharge port. Compared with the prior art, the present invention combines the potential energy of the fluid with the right-angle impact principle to effectively improve the mixing effect, so that the MCF-7 cells can be fully labeled.

[0043] (3) The present invention includes a plurality of microfluidic channels, a plurality of capture claws, and a plurality of diversion channels. The mixed fluid is diverted into a plurality of microfluidic channels through a plurality of diversion channels and continues to flow toward the confluence portion. When the fluid enters the microfluidic channel, the fluid velocity increases due to the decrease in channel width. At this time, the decrease in channel cross-sectional area will lead to an increase in flow velocity under the same flow rate. At the same time, the narrow channel will force the cells in the fluid to gather toward the center line of the channel. At this time, because the shear force in the narrow channel is relatively large, the cells at the edge of the fluid are more easily pushed toward the center line. When the fluid enters the fork, the flow rate will slow down, but at this time the cells have already concentrated near the center line of the channel. A capture claw is provided at the central position of the fork for capturing a single cell. Since the cells have already concentrated near the center line, they are more easily captured by the capture claw. The design of the channel with alternating narrow and wide paths through which the fluid flows makes the fluid accelerate in the narrow channel and focus the cells to the center line; decelerate in the wide channel and capture the cells in the capture claw. This process can be repeated many times to further improve the efficiency of cell capture.

[0044] (4) The present invention includes a plurality of microfluidic channels, a plurality of recesses, and a plurality of screening ports. Through the provision of the plurality of screening ports, the remaining cells can continue to flow out through the screening ports after entering the recesses, thereby screening out the target cells. By providing the specific shapes and sizes of the main microfluidic channels, the plurality of recesses, and the plurality of screening ports and cooperating with ER antibodies, the capture rate of the capture claw is further improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative work.

[0046] Figure 1 is a partial perspective structural schematic diagram of a preferred embodiment of the present invention;

[0047] Figure 2 It is a partial perspective structural diagram of a microfluidic channel of a preferred embodiment of the present invention.

[0048] In the figure: 1. microfluidic chip; 2. first inlet; 3. first feed channel; 4. second inlet; 5. second feed channel; 6. mixing chamber; 7. outlet; 8. microfluidic channel; 801. fork; 802. capture claw; 803. recess; 804. screening port; 9. collecting port; 10. collecting tank. DETAILED DESCRIPTION

[0049] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0050] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein. Therefore, the protection scope of the present invention is not limited to the specific embodiments disclosed below.

[0051] In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the scope of protection of the present application. In addition, the terms "first", "second", etc. are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, features defined as "first", "second", etc. may explicitly or implicitly include one or more of the features. In the description of the invention, unless otherwise specified, "multiple" means two or more.

[0052] In the description of this application, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood by specific circumstances.

[0053] The present invention is applied to capture MCF-7 cells, the diameter of which is between 20-30 μm. The cell flow containing MCF-7 cells also contains peripheral blood mononuclear cells (such as lymphocytes, monocytes, etc.) and red blood cells, wherein the diameter of peripheral blood mononuclear cells is usually between 7-15 μm, which is significantly smaller than that of MCF-7 cells, and the diameter of red blood cells is about 7-8 μm, which is much smaller than that of MCF-7 cells.

[0054] like Figure 1 , Figure 2As shown, a tumor cell counting device based on a microfluidic chip includes: a microfluidic chip 1, an external pump thrust device, a mixing section, a first inlet 2, a second inlet 4, a microfluidic section, a confluence section, and an external collection device. The external pump thrust device is used to control the flow of the reaction liquid between the functional areas of the microfluidic chip 1. The mixing section is arranged in the microfluidic chip 1. The first inlet 2 is arranged on the upper end surface of the microfluidic chip 1 and is connected to the mixing section. The second inlet 4 is arranged on the upper end surface of the microfluidic chip 1 and is connected to the mixing section. The microfluidic section is arranged on one side of the mixing section and is connected to the mixing section. The confluence section is arranged on the side of the microfluidic section away from the mixing section and is connected to the microfluidic section. And an external collection device is arranged above the microfluidic chip 1, which is used to collect the flow and quantity of the microfluidic chip 1.

[0055] Fluid is input into the first inlet 2 and the second inlet 4 of the microfluidic chip 1 respectively through an external pump thrust device. The fluid enters the mixing part and is mixed by its own potential energy. The cells in the mixed fluid mixture are marked and enter the microfluidic part. The microfluidic part captures the target cells in the fluid and finally flows into the confluence part.

[0056] like Figure 1 As shown, the mixing part includes: a mixing chamber 6, a discharge port 7, a first feed channel 3, and a second feed channel 5. The mixing chamber 6 is arranged in the microfluidic chip 1, and its cross section is circular. The discharge port 7 is arranged in the middle of the lower side of the first mixing chamber 6, is connected to the first mixing chamber 6, and its diameter is smaller than the cross-sectional diameter of the first mixing chamber 6. The first feed channel 3 is arranged tangentially along the outer wall of the first mixing chamber 6, and is respectively connected to the first mixing chamber 6 and the first inlet 2. And the second feed channel 5 is arranged tangentially along the outer wall of the first mixing chamber 6, and its direction is perpendicular to the first feed channel 3, and is respectively connected to the first mixing chamber 6 and the second inlet 4, and one end thereof is arranged close to the first feed channel 3.

[0057] The fluid enters the first mixing chamber 6 through the first feed channel 3 and the second feed channel 5 respectively, and the fluid is fully mixed by itself by utilizing the right-angle impact principle, and then the fluid rotates downward to enter the discharge port 7.

[0058] like Figure 1 , Figure 2As shown, the microfluidic part includes: a plurality of microfluidic channels 8, a plurality of capture claws 802, and a plurality of shunt channels. The plurality of microfluidic channels 8 are arranged side by side in the microfluidic chip 1, and the single diameter thereof is 50.0-60.0 μm. A plurality of forks 801 are arranged at intervals in the plurality of microfluidic channels 8, and the single-side diameter of the plurality of forks 801 is the same as the diameter of the microfluidic channels 8. A plurality of capture claws 802 are arranged in the plurality of forks 801, and the side faces thereof are arranged in a streamlined shape. A plurality of capture claws 802 are respectively provided with notches 803 on the side facing the discharge port 7, and the diameter of the notches 803 is 20-30 μm. And a plurality of shunt channels are respectively arranged between the discharge port 7 and the plurality of microfluidic channels 8, and are respectively connected to the discharge port 7 and the plurality of microfluidic channels 8, and the diameters of the plurality of shunt channels are respectively the same as the diameters of the plurality of microfluidic channels 8.

[0059] The capture claw is a microfluidic chip between the two side channels of the fork, and the notch is arc-shaped. The mixed fluid is shunted into several microfluidic channels 8 through several shunt channels, and continues to flow to the confluence. When the fluid enters the microfluidic channel 8, the fluid velocity will increase due to the reduction in channel width. At this time, because the cross-sectional area of ​​the channel is reduced under the same flow rate, the flow rate will increase. At the same time, the narrow channel will force the cells in the fluid to gather toward the center line of the channel. At this time, because the shear force in the narrow channel is large, the cells at the edge of the fluid are more easily pushed to the center line. When the fluid enters the fork 801, the flow rate will slow down, but at this time the cells are already concentrated near the center line of the channel. A capture claw 802 is provided in the central position of the fork 801 for capturing a single cell. Since the cells are already concentrated near the center line, they are more easily captured by the capture claw 802. The channel design with alternating narrow and wide paths for the fluid flow accelerates the fluid in the narrow channel and focuses the cells to the center line. Slow down in the wide channel and capture cells in the capture claw 802. This process can be repeated many times to further improve the capture efficiency of cells.

[0060] like Figure 1 , Figure 2 As shown, the microfluidic portion includes: a plurality of screening ports 804, which are respectively disposed in the plurality of capturing claws 802, respectively connected to the plurality of recesses 803, and penetrate the plurality of capturing claws 802, and the diameter of the plurality of screening ports 804 is 12-18 μm.

[0061] By setting a plurality of screening ports 804 , the remaining cells can continue to flow out through the screening ports 804 after entering the recess 803 , thereby screening out the target cells and further improving the capture rate of the capture claw 802 .

[0062] like Figure 1As shown, the confluence part includes: a plurality of confluence channels and a collecting port 9. The plurality of confluence channels are respectively arranged on the side of the plurality of microfluidic channels 8 away from the discharge port 7, and are respectively connected with the plurality of microfluidic channels 8. The collecting port 9 is arranged on the upper end surface of the microfluidic chip 1, and is connected with the plurality of confluence channels.

[0063] The fluids flowing through the plurality of microfluidic channels 8 are respectively collected into the manifold 9 through the plurality of converging channels, so as to ensure that the fluids in the plurality of microfluidic channels 8 can flow smoothly.

[0064] The recesses 803 of several of the capture claws 802 are respectively coated with ER antibodies.

[0065] The surface of the capture claw 802 is coated with ER antibodies that specifically bind to MCF-7 cells. ER is highly expressed in MCF-7 cells. MCF-7 cells are an estrogen receptor-positive breast cancer cell line with a high expression of estrogen receptor (ER) in the cells. ER antibodies can specifically recognize and bind to these ERs, thereby capturing MCF-7 cells. ER antibodies have high specificity and can accurately recognize and bind to ERs without binding to other non-specific proteins. This specificity enables ER antibodies to effectively reduce interference with other non-target cells when capturing MCF-7 cells, thereby improving the accuracy and efficiency of capture.

[0066] The external acquisition device includes: a data processor, a flow sensor, and an image collector. The flow sensor is arranged at the discharge port 7 and is electrically connected to the data processor; and the image collector is arranged above the microfluidic channels 8 and is connected to the data processor.

[0067] By setting up the flow sensor, the fluid flow through the discharge port 7 can be measured in real time, and the data can be transmitted to the data processor. The number of captured cells can be observed through the image acquisition device and the data can be transmitted to the data processor. The data is processed and the density and content of the target cells in the fluid can be obtained based on the transmission data, the established volume data of several diversion channels, the capture rate and the count.

[0068] like Figure 1 As shown, it includes: a collecting tank 10, which is arranged on the upper end surface of the microfluidic chip 1 and is connected to the collecting port 9.

[0069] By providing the collecting groove 10 , the volume of the fluid flowing through the plurality of microfluidic channels 8 can be increased while preventing the fluid from flowing out of the microfluidic chip to cause experimental contamination.

[0070] The material of the microfluidic chip 1 is PDMS.

[0071] PDMS has good transparency, biocompatibility, non-toxicity and gas permeability, and is suitable for bio-based microfluidic applications, such as cell experiments, etc. However, it has a strong adsorption capacity for nonspecific molecules, which may lead to the absorption of organic solvents and proteins.

[0072] A method for using a tumor cell counting device based on a microfluidic chip, comprising:

[0073] Step 1: First, a cell flow containing MCF-7 cells and a fluorescent agent are respectively input into the first inlet 2 and the second inlet 4 through an external inference device, and mixed in the microfluidic chip 1 to obtain a mixed solution;

[0074] Step 2: The mixed solution is split and introduced into the microfluidic channels 8 with different narrow and wide alternations, so that the cells are focused on the center line of the fluid and captured by the capture claws 802;

[0075] Step 3: Collect the image of the microfluidic channel 8 and count according to the image;

[0076] Step 4: Based on the capture rate and counts, the actual tumor content is obtained.

[0077] The tumor cell counting device based on the microfluidic chip of the present invention is used to test the device multiple times in combination with a cell flow with a known number of MCF-7 cells, and the capture rate of the device is obtained by using the capture number and the number of MCF-7 cells.

[0078] In Example 1, the diameter of the plurality of microfluidic channels is 50 μm, the diameter of the plurality of recesses is 20 μm, and the diameter of the plurality of screening ports is 12 μm;

[0079] In Example 2, the diameter of the plurality of microfluidic channels is 55 μm, the diameter of the plurality of recesses is 20 μm, and the diameter of the plurality of screening ports is 12 μm;

[0080] In Example 3, the diameter of the plurality of microfluidic channels is 60 μm, the diameter of the plurality of recesses is 20 μm, and the diameter of the plurality of screening ports is 12 μm;

[0081] In Example 4, the diameter of the plurality of microfluidic channels is 50 μm, the diameter of the plurality of recesses is 25 μm, and the diameter of the plurality of screening ports is 12 μm;

[0082] In Example 5, the diameter of the plurality of microfluidic channels is 50 μm, the diameter of the plurality of recesses is 30 μm, and the diameter of the plurality of screening ports is 12 μm;

[0083] In Example 6, the diameter of the plurality of microfluidic channels is 50 μm, the diameter of the plurality of recesses is 20 μm, and the diameter of the plurality of screening ports is 15 μm;

[0084] In Example 7, the diameter of the plurality of microfluidic channels is 50 μm, the diameter of the plurality of recesses is 20 μm, and the diameter of the plurality of screening ports is 18 μm;

[0085] Table 1 MCF-7 cell capture rate change table

[0086]

[0087]

[0088] The above experimental table indicates that the MCF-7 cell capture rate is the highest when the diameters of the microfluidic channels are all 55 μm, the diameters of the recesses are all 25 μm, and the diameters of the screening ports are all 15 μm:

[0089] Microfluidic channel diameter / μm Notch diameter / μm Screening port diameter / μm Capture rate / % 55 25 15 95.0

[0090] When the present invention is used, the cell flow containing MCF-7 cells and the fluorescent agent are respectively input into the first inlet 2 and the second inlet 4 by the external pump thrust device, and the cell flow and the fluorescent agent enter the mixing chamber 6 through the first feed channel 3 and the second feed channel 5 respectively. The fluid is fully mixed by itself by the right-angle impact principle, and then the fluid rotates downward to enter the discharge port 7, and is diverted to several microfluidic channels 8 through several diversion channels. When the fluid enters the diversion channel, the fluid velocity will increase due to the reduction in channel width. At this time, because the cross-sectional area of ​​the channel is reduced at the same flow rate, the flow rate will increase. At the same time, the narrow channel will force the cells in the fluid to gather toward the center line of the channel. At this time, because the shear force in the narrow channel is large, the cells at the edge of the fluid are more easily pushed to the center line. When the fluid enters the fork 801, the flow rate will slow down, but at this time the cells are already concentrated near the center line of the channel. A capture claw 802 is provided at the central position of the fork 801 for capturing single cells. Since the cells are already concentrated near the center line, they are more easily captured by the capture claw 802. The design of the alternating narrow and wide channels through which the fluid flows makes the fluid accelerate in the narrow channel and focus the cells to the center line, decelerate in the wide channel, and capture the cells in the capture claw 802. Through the setting of the flow sensor, the fluid flow through the discharge port 7 can be measured in real time, and the data can be transmitted to the data processor, the number of captured cells can be observed through the image collector and the data can be transmitted to the data processor, and the data processing and the density and content of the target cells in the fluid can be obtained based on the transmitted data, the established volume data of several diversion channels, the capture rate and the count.

[0091] The above is based on the ideal embodiment of the present invention. Through the above description, relevant personnel can make various changes and modifications without departing from the technical concept of the present invention. The technical scope of the present invention is not limited to the content in the specification, and the technical scope must be determined according to the scope of the claims.

Claims

1. A tumor cell counting device based on a microfluidic chip, comprising: Microfluidic chip (1), An external pump thrust device is used to control the flow of the reaction liquid between the functional areas of the microfluidic chip (1); A mixing unit, arranged in the microfluidic chip (1); A first inlet (2) is arranged on the upper end surface of the microfluidic chip (1) and is connected to the mixing part; A second inlet (4) is arranged on the upper end surface of the microfluidic chip (1) and is connected to the mixing part; A microfluidic section, disposed on one side of the mixing section and communicated with the mixing section; A confluence portion, disposed on a side of the microfluidic portion away from the mixing portion and communicated with the microfluidic portion; as well as An external collection device is arranged above the microfluidic chip (1) and is used to collect flow and quantity of the microfluidic chip (1).

2. The tumor cell counting device based on a microfluidic chip according to claim 1, characterized in that: The mixing unit comprises: A mixing chamber (6) is arranged in the microfluidic chip (1) and has a circular cross section; a discharge port (7), which is arranged at the middle of the lower side of the first mixing chamber (6), is communicated with the first mixing chamber (6), and has a diameter smaller than the cross-sectional diameter of the first mixing chamber (6); a first feed channel (3), arranged tangentially along the outer wall of the first mixing chamber (6), and communicating with the first mixing chamber (6) and the first inlet (2) respectively; and The second feed channel (5) is arranged tangentially along the outer wall of the first mixing chamber (6) and in a direction perpendicular to the first feed channel (3). It is connected to the first mixing chamber (6) and the second inlet (4) respectively, and one end of the second feed channel (5) is arranged close to the first feed channel (3).

3. The tumor cell counting device based on a microfluidic chip according to claim 2, characterized in that: The microfluidic section comprises: A plurality of microfluidic channels (8) are arranged side by side in the microfluidic chip (1), each of which has a diameter of 50.0-60.0 μm, and a plurality of branch channels (801) are arranged at intervals in the plurality of microfluidic channels (8), and the diameter of a single side of the plurality of branch channels (801) is the same as the diameter of the microfluidic channel (8); A plurality of capturing claws (802) are respectively arranged in the plurality of forked channels (801), and the side surfaces thereof are arranged in a streamlined shape. A plurality of capturing claws (802) are respectively provided with notches (803) on the side facing the discharge port (7), and the diameter of the notches (803) is 20-30 μm; and A plurality of flow diversion channels are respectively arranged between the discharge port (7) and the plurality of microfluidic channels (8), and are respectively connected to the discharge port (7) and the plurality of microfluidic channels (8); and the diameters of the plurality of flow diversion channels are respectively the same as the diameters of the plurality of microfluidic channels (8).

4. The tumor cell counting device based on a microfluidic chip according to claim 4, characterized in that: The microfluidic section comprises: A plurality of screening ports (804) are respectively arranged in the plurality of capturing claws (802), are respectively connected to the plurality of recesses (803), and penetrate the plurality of capturing claws (802), and the diameter of the plurality of screening ports (804) is 12-18 μm.

5. The tumor cell counting device based on a microfluidic chip according to claim 3, characterized in that: The confluence portion comprises: A plurality of converging channels, respectively arranged on a side of the plurality of microfluidic channels (8) away from the discharge port (7), and respectively connected to the plurality of microfluidic channels (8); and A flow collecting port (9) is arranged on the upper end surface of the microfluidic chip (1) and is connected to a plurality of the flow collecting channels.

6. The tumor cell counting device based on a microfluidic chip according to claim 3, characterized in that: The recesses (803) of the plurality of capture claws (802) are coated with ER antibodies respectively.

7. The tumor cell counting device based on a microfluidic chip according to claim 3, characterized in that: The external acquisition device comprises: Data processors; a flow sensor, disposed at the discharge port (7) and electrically connected to the data processor; and An image collector is arranged above the plurality of microfluidic channels (8) and is connected to a data processor.

8. The tumor cell counting device based on a microfluidic chip according to claim 5, characterized in that: include: A collecting tank (10) is arranged on the upper end surface of the microfluidic chip (1) and is connected to the collecting port (9).

9. The tumor cell counting device based on a microfluidic chip according to claim 1, characterized in that: The material of the microfluidic chip (1) is PDMS.

10. A method for using a tumor cell counting device based on a microfluidic chip according to any one of claims 1 to 9, characterized in that: Step 1: First, a cell flow containing MCF-7 cells and a fluorescent agent are respectively input into the first inlet (2) and the second inlet (4) through an external inference device, and mixed in the microfluidic chip (1) to obtain a mixed solution; Step 2: The mixed solution is split and introduced into microfluidic channels (8) with different narrow and wide alternations, so that the cells are focused on the center line of the fluid and captured by the capture claws (802); Step 3: Collecting an image of the microfluidic channel (8) and counting based on the image; Step 4: Based on the capture rate and counts, the actual tumor content is obtained.

Images