A microfluidic chip rapid detection system

Abstract

The present invention relates to the technical field of cell detection, and specifically to a microfluidic chip rapid detection system, comprising a sample pretreatment and fluid control module, which is used to perform preliminary treatment on input sample cells and ensure that the samples flow accurately in the microfluidic chip; a sensor detection and signal processing module, which is used to perform real-time detection on the sample cells after pretreatment, and send the detected data signals to a circulating tumor cell capture module; the circulating tumor cell capture module is used to capture circulating tumor cells according to the data signals; the present invention integrates optical sensors with microfluidic channels to realize real-time detection of optical signals and fluorescence signals of sample cells, and can more accurately extract circulating tumor cell characteristics; by introducing magnetic field manipulation technology and multi-channel confined microstructure design, combined with fluid dynamic regulation of shear stress, directional capture of circulating tumor cells is achieved.

Description

Technical Field

[0001] The invention relates to the technical field of cell detection, in particular to a microfluidic chip rapid detection system. Background Art

[0002] The detection of circulating tumor cells is of great significance for the early diagnosis of cancer, efficacy evaluation, recurrence monitoring and prognosis judgment. However, the current CTC detection technology still faces multiple challenges.

[0003] In existing microfluidic systems, the optical detection signals for circulating tumor cells are relatively single and cannot fully utilize multidimensional signals for high-precision detection, resulting in misjudgment or omission of some cells;

[0004] When traditional capture technology is used to capture circulating tumor cells in a targeted manner, the target cells are easily lost or captured inaccurately, making it difficult to achieve efficient targeted capture.

[0005] The fluid control accuracy of microfluidic chips in the prior art is insufficient, and sample blockage may occur easily due to flow rate fluctuations or excessive flow rate may affect the detection and capture effects. Summary of the invention

[0006] In view of the above-mentioned problems, the present invention proposes a microfluidic chip rapid detection system, which can effectively solve the problem that the traditional detection method proposed in the above background technology has a single detection means and is difficult to simultaneously obtain multi-dimensional optical information of cells, which easily leads to damage or omission of circulating tumor cells during the detection process.

[0007] In order to solve the above technical problems, the present invention provides the following technical solutions: a microfluidic chip rapid detection system, comprising:

[0008] The sample pre-processing and fluid control module is used to perform preliminary processing on the input sample cells, and is characterized by comprising a sample dilution and separation unit and a fluid drive and control unit, specifically:

[0009] The sample blood is subjected to sample pretreatment, sample separation, and dilution treatment from its original state to form a sample fluid to be detected, and the sample fluid is made to flow into a microfluidic chip;

[0010] And, the sensing detection and signal processing module is used to detect the sample cells and send the detected data signals to the circulating tumor cell capture module, including an optical sensor detection unit, a signal data processing unit and a data transmission unit, specifically:

[0011] An optical sensor is integrated in the microfluidic channel, and a light source is used to illuminate the sample cells to generate discrete light signals and fluorescence signals. The circulating tumor cell features are extracted based on the generated discrete light signals and fluorescence signals, thereby detecting the circulating tumor cells, and sending the detection results to the circulating tumor cell capture module;

[0012] Also includes:

[0013] The circulating tumor cell capture module is used to capture circulating tumor cells according to the data signal, including a circulating tumor cell positioning unit, a circulating tumor cell capture unit and a circulating tumor cell confinement unit, specifically:

[0014] The screened circulating tumor cells are captured in a targeted manner based on magnetic field manipulation technology and multi-channel single-cell confined microstructure design, combined with a microfluidic system to achieve precise cell capture.

[0015] As a preferred solution of the microfluidic chip rapid detection system described in the present invention, the sample pretreatment is specifically as follows:

[0016] Set the input biological blood sample to , and including sample cells And impurities , then there is, The total number of sample cells is ;

[0017] The blood sample is filtered using a membrane to remove impurities.

[0018] in, represents the sample cells in the blood sample, Indicates impurities in blood samples, It indicates the proportion of sample cells in the blood sample after filtration, which is used to control the degree of cell filtration and satisfies the formula , Represents the proportion coefficient of sample cells;

[0019] Update the data of the filtered sample cells, then we have, , Indicates impurities after filtration, Represents a blood sample after impurities have been filtered out.

[0020] As a preferred solution of the microfluidic chip rapid detection system described in the present invention, the sample dilution is specifically implemented as follows:

[0021] After the sample cells are separated, the total number of sample cells is The volume of the sample solution is , the sample cells are at the bottom of the centrifuge tube, and the concentration of the sample cells is Then,

[0022] in, Represents the total number of sample cells, represents the volume of sample solution, Indicates the concentration of sample cells after separation;

[0023] By adding the volume of buffer solution, the initial sample volume Dilution is performed to reduce the viscosity of the sample cells and ensure smooth flow of the fluid in the microfluidic chip.

[0024] in, Represents the total number of sample cells, represents the volume of sample solution, represents the concentration of sample cells after adding buffer, represents the volume of buffer added;

[0025] By adding buffer, the viscosity of the sample cells can be adjusted, then,

[0026] in, represent the initial viscosity of the sample solution and the viscosity of the buffer solution, respectively. represents the volume of sample solution, represents the volume of buffer added, represents the viscosity of sample cells after adjustment;

[0027] The dilution of the sample cells is controlled by monitoring the concentration and viscosity of the sample cells after adding the buffer, as follows:

[0028] The objective function is set based on the sample cell concentration and viscosity after adding the buffer solution, and the objective function is set based on the volume of the buffer solution added. For optimization, we have

[0029] in, Indicates that the sample cells meet the minimum concentration requirement for microfluidic chip detection. represents the maximum viscosity at which sample cells can flow into the microfluidic channel, Represents the objective function. By limiting the result of the objective function, the volume of the added buffer is optimized. Then,

[0030] for The result satisfies the formula as well as , the buffer volume corresponding to the objective function at this time is the volume of buffer added.

[0031] As a preferred solution of the microfluidic chip rapid detection system of the present invention, the method of irradiating sample cells with a light source to generate discrete light signals and fluorescent signals is as follows:

[0032] Discrete optical signal acquisition,

[0033] in, represents the initial intensity of the incident light, represents the cell radius, represents the distance from the cell to the optical sensor, represents the influencing factor function, represents the scattering angle, represents the wavelength of light of the optical sensor, represents the refractive index of the cells in the sample cell, represents the acquired discrete optical signal;

[0034] Fluorescence signal acquisition,

[0035] in, represents the initial intensity of the incident light, represents the fluorescence quantum yield, represents the fluorescence absorbance, represents the concentration of fluorescent dye, represents the acquired fluorescence signal.

[0036] As a preferred solution of the microfluidic chip rapid detection system of the present invention, the detection of circulating tumor cells is specifically as follows:

[0037] According to the extracted circulating tumor cell characteristics, a circulating tumor cell scoring model is constructed, then,

[0038] in, represents the forward scattering intensity, represents the side scattering intensity, represents the relative fluorescence intensity, represents the bias term, Respectively represent the weight coefficient of each feature;

[0039] Screening of circulating tumor cells was performed based on the scoring results, as follows:

[0040] The score of the sample cells under the circulating tumor cell scoring model is , the score of normal cells in the circulating tumor cell scoring model is , the score of circulating tumor cells under the circulating tumor cell scoring model is , according to the score, circulating tumor cells are screened, then,

[0041] Normal cells and circulating tumor cells are screened according to the scoring results of sample cells under the circulating tumor cell scoring model;

[0042] If the score of the sample cell in the circulating tumor cell scoring model satisfies the formula , indicating that the sample cells corresponding to the current score are a collection of circulating tumor cells and normal cells, and the current sample cells need to be screened a second time.

[0043] As a preferred solution of the microfluidic chip rapid detection system of the present invention, the secondary screening is specifically as follows:

[0044] According to the characteristics of the constructed data, a dynamic screening mechanism is constructed.

[0045] Scoring mechanism based on behavioral data characteristics, ,

[0046] Scoring mechanism under spectral data,

[0047] in, represents the movement speed of sample cells in the microfluidic channel, represents the velocity change rate of the sample cell in the microfluidic channel, represents the acceleration of the sample cell in the microfluidic channel, represents the fluorescence response at different wavelengths, Indicates the upper limit of wavelength types, Indicates the type index of wavelength, Indicates the degree of influence of spectral characteristics at different wavelengths on the score. Respectively represent the influence of speed, acceleration and speed change rate on the score. Indicates the scoring result under the behavior data characteristics, Indicates the scoring results under spectral data;

[0048] Combining the scoring models under the two data characteristics to construct a comprehensive scoring model, we have:

[0049] in, They represent the weight coefficients of behavioral scoring and spectral scoring respectively, which are set by the implementers according to the actual application scenarios. Indicates the scoring result under the behavior data characteristics, Indicates the scoring results under spectral data;

[0050] Screening is performed based on behavioral scores, spectrum scores, and comprehensive score models, as follows:

[0051] Using normal cells to set behavioral scoring thresholds , spectral scoring standard threshold , and the comprehensive scoring criteria threshold ,

[0052] Screening of circulating tumor cells in sample cells was performed according to the set standard threshold.

[0053] As a preferred solution of the microfluidic chip rapid detection system of the present invention, the circulating tumor cell localization unit is specifically as follows:

[0054] Through magnetic labeling technology, magnetic particles are attached to the surface of circulating tumor cells. Under the action of , circulating tumor cells move along the magnetic field lines, then,

[0055] in, represents the magnetic moment of magnetic particles attached to the surface of circulating tumor cells, express The magnetic field strength at the moment, represents the initial magnetic field strength, Indicates the control coefficient, which is used to adjust the sensitivity of magnetic field changes. It is set by the implementer according to the actual application scenario. represents the real-time displacement difference of circulating tumor cells, Represents the magnetic field force, acting on magnetic particles, used to guide the movement of circulating tumor cells.

[0056] As a preferred solution of the microfluidic chip rapid detection system of the present invention, the directional capture of the screened circulating tumor cells is specifically as follows:

[0057] By controlling the flow rate in each channel, a differentiated fluid environment is generated, and the dynamic regulation of shear stress is achieved, then,

[0058] in, represents the viscosity of the fluid, represents the velocity difference of the flow velocity in the channel, represents the distance between fluid layers in the channel, represents dynamically regulated shear stress;

[0059] Monitoring the deformation of circulating tumor cells based on dynamically regulated shear stress combined with real-time imaging , according to the monitored deformation of circulating tumor cells, the directional capture of circulating tumor cells is achieved, then,

[0060] in, represents the shape of circulating tumor cells, represents the dynamically regulated shear stress, represents the elastic modulus of circulating tumor cells, represents the velocity coefficient of circulating tumor cells responding to shear stress, Indicates the time of shear stress action;

[0061] Different cell capture devices are designed at the end of each microfluidic channel. The deformation of circulating tumor cells in the channel is determined according to the shear stress in different channels. Different, by setting the standard threshold of deformation of circulating tumor cells under different shear stresses ,If the shape variable of circulating tumor cells reaches the standard shape variable threshold, the current circulating tumor cells are guided into a specific area for directional capture.

[0062] Beneficial effects of the present invention: The present invention integrates optical sensors with microfluidic channels to realize real-time detection of optical signals and fluorescence signals of sample cells, which can more accurately extract the characteristics of circulating tumor cells and avoid damage to cells during the detection process; by introducing magnetic field manipulation technology and multi-channel confined microstructure design, combined with dynamic adjustment of shear stress by fluid, the directional capture of circulating tumor cells is effectively achieved with high capture accuracy and no cell loss or escape; by introducing a comprehensive scoring model, multi-parameter analysis of detected cells is performed to achieve automatic screening; through real-time imaging technology and feedback mechanism, the shear stress and capture conditions are dynamically adjusted to further improve the capture efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. Among them:

[0064] Figure 1 The figure is a schematic diagram of the overall structure of a microfluidic chip rapid detection system of the present invention.

[0065] Figure 2 The present invention is a schematic diagram of the logic flow of circulating tumor cell screening in a microfluidic chip rapid detection system. DETAILED DESCRIPTION

[0066] In order to make the above-mentioned purposes, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the drawings of the specification. Obviously, the described embodiments are part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary persons in the art without creative work should fall within the scope of protection of the present invention.

[0067] 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, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0068] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0069] The present invention is described in detail with reference to schematic diagrams. When describing the embodiments of the present invention, for the sake of convenience, the cross-sectional diagrams showing the device structure will not be partially enlarged according to the general scale, and the schematic diagrams are only examples, which should not limit the scope of protection of the present invention. In addition, in actual production, the three-dimensional dimensions of length, width and depth should be included.

[0070] Meanwhile, in the description of the present invention, it should be noted that the terms "first, second or third" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0071] In the present invention, unless otherwise clearly specified and limited, the terms "install, connect, connect" should be understood in a broad sense, for example: it can be a fixed connection, a detachable connection or an integral connection; it can also be a mechanical connection, an electrical connection or a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0072] Example 1: Reference Figure 1-2 , as one embodiment of the present invention, provides a microfluidic chip rapid detection system, including a sample pretreatment and fluid control module, a sensor detection and signal processing module, and a circulating tumor cell capture module;

[0073] Specifically, the sample pre-processing and fluid control module is used to perform preliminary processing on the input sample cells and ensure that the sample flows accurately in the microfluidic chip;

[0074] The sensor detection and signal processing module is used to perform real-time detection on the sample cells after pretreatment, and send the detected data signals to the circulating tumor cell capture module;

[0075] The circulating tumor cell capture module is used to capture circulating tumor cells according to data signals.

[0076] It should be noted that the entire system inputs sample cells into the microfluidic chip through the collaboration of three modules. The sample pretreatment and fluid control module pretreatments the input sample cells and controls the flow of samples in the microfluidic chip. Then the sensor detection and signal processing module performs real-time detection of the pretreated sample cells and sends the detection results to the circulating tumor cell capture module, which captures circulating tumor cells based on the detected data signals. Through teamwork between modules, data transmission and operation steps are reduced, and the efficiency and accuracy of the entire system are improved.

[0077] Furthermore, the sample preprocessing and fluid control module performs data preprocessing on the sample cells input into the microfluidic chip and controls the flow rate of the sample cells in the microfluidic chip, including a sample dilution and separation unit and a fluid drive and control unit. The sample dilution and separation unit processes the blood from its original state through the steps of sample preprocessing, sample separation, and dilution processing to ensure that the sample cells can enter the microfluidic chip for subsequent capture and detection.

[0078] Specifically, the sample pretreatment is to initially remove impurities from the blood sample to ensure that the sample cells can enter the microfluidic chip, as follows:

[0079] Set the input biological blood sample to , and including sample cells And impurities , then there is, The total number of sample cells is ;

[0080] The blood sample is filtered using a membrane to remove impurities.

[0081] in, represents the sample cells in the blood sample, Indicates impurities in blood samples, It indicates the proportion of sample cells in the blood sample after filtration, which is used to control the degree of cell filtration and satisfies the formula , Represents the proportion coefficient of sample cells;

[0082] Update the data of the filtered sample cells, then we have, , Indicates impurities after filtration, Represents a blood sample after impurities have been filtered out.

[0083] The sample separation is to separate the sample cells by centrifugation technology, as follows:

[0084] According to the radius and density of the sample cells, the separation of sample cells and impurities is achieved, then,

[0085]

[0086] in, represents the radius of the sample cell, Represent the density of sample cells and the density of blood sample, respectively. represents the centrifugal speed, Indicates the viscosity of the blood sample medium, which is controlled by the characteristics of the blood sample. Indicates the sedimentation velocity. The sedimentation velocity of the sample cells is adjusted by the radius and density of the sample cells. Greater than the sedimentation rate of impurities , so that the sample cells are preferentially settled to the bottom of the centrifuge tube, and the impurities are located in the upper layer of the centrifuge tube.

[0087] The sample dilution is achieved by reducing the viscosity of the blood sample to ensure smooth flow of the fluid in the microfluidic channel, which is specifically implemented as follows:

[0088] After the sample cells are separated, the total number of sample cells is The volume of the sample solution is , the sample cells are at the bottom of the centrifuge tube, and the concentration of the sample cells is Then,

[0089] in, Represents the total number of sample cells, represents the volume of sample solution, Indicates the concentration of sample cells after separation;

[0090] By adding the volume of buffer solution, the initial sample volume Dilution is performed to reduce the viscosity of the sample cells and ensure smooth flow of the fluid in the microfluidic chip.

[0091] in, Represents the total number of sample cells, represents the volume of sample solution, represents the concentration of sample cells after adding buffer, represents the volume of buffer added;

[0092] By adding buffer, the viscosity of the sample cells can be adjusted, then,

[0093] in, represent the initial viscosity of the sample solution and the viscosity of the buffer solution, respectively. represents the volume of sample solution, represents the volume of buffer added, represents the viscosity of sample cells after adjustment;

[0094] The dilution of the sample cells is controlled by monitoring the concentration and viscosity of the sample cells after adding the buffer, as follows:

[0095] The objective function is set based on the sample cell concentration and viscosity after adding the buffer solution, and the objective function is set based on the volume of the buffer solution added. For optimization, we have

[0096] in, Indicates that the sample cells meet the minimum concentration requirement for microfluidic chip detection. represents the maximum viscosity at which sample cells can flow into the microfluidic channel, Represents the objective function. By limiting the result of the objective function, the volume of the added buffer is optimized. Then,

[0097] for The result satisfies the formula as well as When , the buffer volume corresponding to the objective function at this time is the volume of buffer added, which can ensure the subsequent capture of sample cells while ensuring that the sample can pass through the microfluidic chip smoothly to avoid fluid blockage.

[0098] The fluid drive and control unit realizes the flow of sample cells in the microfluidic chip by combining pressure drive with electric fluid regulation, and can accurately control and maintain a stable fluid environment. The specific implementation is as follows:

[0099] Generates stable pressure with a micro pump , provides flow rate for sample fluid and pushes sample fluid into microfluidic channel, then,

[0100]

[0101] in, Indicates that the micro pump is The pressure provided at all times, Indicates that the sample fluid is The flow rate of time, represents the density of the sample fluid;

[0102] Set the standard speed for sample fluid to enter the microfluidic channel stably By monitoring the real-time flow rate of the sample fluid and adjusting the pressure feedback, the flow of sample cells in the microfluidic chip can be ensured.

[0103] When the real-time velocity of the sample fluid is compared with the standard velocity, the formula is satisfied. When , it means that the current sample flow rate exceeds the standard speed, and the pressure provided by the micro pump is reduced to achieve a reduction in the sample fluid speed, thereby completing the precise control of the sample flow rate;

[0104] When the real-time velocity of the sample fluid is compared with the standard velocity, the formula is satisfied. When , it means that the current sample flow rate is lower than the standard speed, and the pressure provided by the micro pump is increased to increase the sample fluid speed, thereby completing the precise control of the sample flow rate;

[0105] When the real-time velocity of the sample fluid is compared with the standard velocity, the formula is satisfied. When , it means that the current sample flow rate is equal to the standard speed, maintaining the pressure provided by the current micropump to ensure that the sample fluid stably enters the microfluidic chip.

[0106] Furthermore, the sensor detection and signal processing module is used to realize real-time detection of sample cells input into the microfluidic channel, and send the detected data signals to the circulating tumor cell capture module for subsequent capture of circulating tumor cells. The sensor detection and signal processing module includes an optical sensor detection unit, a signal data processing unit and a data transmission unit. The optical sensor detection unit is responsible for real-time optical detection of sample cells and generating data, the signal data processing unit is responsible for processing the detected data and extracting relevant features of circulating tumor cells, and the data transmission unit sends the extracted target cell features to the circulating tumor cell capture module for subsequent capture of the target cells.

[0107] Specifically, the optical sensor detection unit integrates an optical sensor in a microfluidic channel to perform real-time detection of cells flowing through the channel. When circulating tumor cells pass through the optical detection area, the light emitted by the light source irradiates the cells and generates two signals at the same time, including a discrete light signal and a fluorescence signal. Then,

[0108] Discrete optical signal acquisition,

[0109] in, represents the initial intensity of the incident light, represents the cell radius, represents the distance from the cell to the optical sensor, represents the influencing factor function, represents the scattering angle, represents the wavelength of light of the optical sensor, represents the refractive index of the cells in the sample cell, represents the acquired discrete optical signal;

[0110] Fluorescence signal acquisition,

[0111] in, represents the initial intensity of the incident light, represents the fluorescence quantum yield, represents the fluorescence absorbance, represents the concentration of fluorescent dye, represents the acquired fluorescence signal.

[0112] The signal data processing unit processes the signal collected by the optical sensor so as to detect circulating tumor cells. The specific processing is as follows:

[0113] Based on the collection of discrete optical signals And fluorescent signal Complete the extraction of circulating tumor cell characteristics, including forward scatter intensity, side scatter intensity and fluorescence relative intensity, then we have:

[0114] Forward scatter intensity used to reflect cell volume ,

[0115]

[0116] Side scatter intensity, used to reflect the internal structure and complexity of cells ,

[0117]

[0118] Relative fluorescence intensity ,

[0119]

[0120] in, represents the collected forward scattering intensity, represents the collected side scatter intensity, represents the intensity of the collected fluorescence signal. represents the forward scatter conversion factor of the optical sensor, represents the side scatter conversion factor of the optical sensor, The conversion factor representing the fluorescence signal, the value of the conversion factor is determined by the implementer according to the actual correction result of the optical sensor in practical application, and will not be described in detail in the present invention;

[0121] According to the extracted circulating tumor cell characteristics, a circulating tumor cell scoring model is constructed, then,

[0122] in, represents the forward scattering intensity, represents the side scattering intensity, represents the relative fluorescence intensity, represents the bias term, They represent the weight coefficients of each feature, which are set by the implementers according to the actual application scenarios.

[0123] Furthermore, the detection of circulating tumor cells is conducive to constructing a circulating tumor cell scoring model to score the input sample cells, and screening circulating tumor cells according to the scoring results, as follows:

[0124] The sample cells input into the microfluidic channel are used as the input of the circulating tumor cell scoring model. A normal cell and a circulating tumor cell are prepared as a comparison group. The output results of the circulating tumor cell scoring model are used as the screening basis for circulating tumor cells. Then,

[0125] The score of the sample cells under the circulating tumor cell scoring model is , the score of normal cells in the circulating tumor cell scoring model is , the score of circulating tumor cells under the circulating tumor cell scoring model is , according to the score, circulating tumor cells are screened, then,

[0126] If the score of the sample cell in the circulating tumor cell scoring model satisfies the formula , indicating that the sample cell corresponding to the current score is a normal cell;

[0127] If the score of the sample cell in the circulating tumor cell scoring model satisfies the formula , indicating that the sample cells corresponding to the current score are circulating tumor cells;

[0128] If the score of the sample cell in the circulating tumor cell scoring model satisfies the formula , indicating that the sample cells corresponding to the current score are a collection of circulating tumor cells and normal cells, and the current sample cells need to be screened a second time.

[0129] Furthermore, the secondary screening is performed on sample cells in the form of aggregates composed of circulating tumor cells and normal cells by constructing a dynamic screening mechanism based on multi-parameter comprehensive scoring. The dynamic screening mechanism based on multi-parameter comprehensive scoring combines the dynamic behavior analysis of cells and multi-wavelength spectral analysis to improve the screening accuracy of circulating tumor cells. The specific screening is as follows:

[0130] Using real-time imaging technology to track the movement of sample cells in the microfluidic channel, record the movement trajectory, speed and acceleration, and construct behavioral data features, we have:

[0131] in, represents the movement speed of sample cells in the microfluidic channel, represents the velocity change rate of the sample cell in the microfluidic channel, represents the acceleration of the sample cell in the microfluidic channel;

[0132] And collect the fluorescence intensity and scattering signal intensity at different wavelengths, and construct the spectral data characteristics at the same time, then we have,

[0133]

[0134] Represents the scattered light intensity at different wavelengths, represents the fluorescence intensity at different wavelengths, represents the light absorption signal, Indicates that the wavelength is Fluorescence response under

[0135] According to the characteristics of the constructed data, a dynamic screening mechanism is constructed.

[0136] Scoring mechanism based on behavioral data characteristics,

[0137]

[0138] Scoring mechanism under spectral data,

[0139]

[0140] in, represents the movement speed of sample cells in the microfluidic channel, represents the velocity change rate of the sample cell in the microfluidic channel, represents the acceleration of the sample cell in the microfluidic channel, represents the fluorescence response at different wavelengths, Indicates the upper limit of wavelength types, Indicates the type index of wavelength, Indicates the degree of influence of spectral characteristics at different wavelengths on the score. Respectively represent the influence of speed, acceleration and speed change rate on the score. Indicates the scoring result under the behavior data characteristics, Represents the scoring results under spectral data.

[0141] The combined cell dynamic behavior analysis and multi-wavelength spectral analysis is to combine the scoring models under the two data characteristics to construct a comprehensive scoring model, then,

[0142] in, They represent the weight coefficients of behavioral scoring and spectral scoring respectively, which are set by the implementers according to the actual application scenarios. Indicates the scoring result under the behavior data characteristics, Represents the scoring results under spectral data.

[0143] The secondary screening of sample cells is based on behavioral scores, spectral scores and comprehensive scoring models, as follows: using normal cells to set behavioral scoring standard thresholds , spectral scoring standard threshold , and the comprehensive scoring criteria threshold ,

[0144] According to the set standard threshold, the sample cells are screened twice, then:

[0145] If the score of the sample cell under the comprehensive scoring model is compared with the standard threshold, the result satisfies the formula , the current sample cells are scored spectrally and behaviorally respectively. If the comparison results satisfy the formula and , it means that the current sample cells are circulating tumor cells;

[0146] If the score of the sample cell under the comprehensive scoring model is compared with the standard threshold, the result satisfies the formula , the current sample cells are scored spectrally and behaviorally respectively. If the comparison results satisfy the formula or When the current sample cells are input into the circulating tumor cell scoring model for re-scoring, if the screening of the current sample cells is performed to the comprehensive scoring model, and the scoring result of the comprehensive scoring model is consistent with the previous time, it means that the current sample cells are non-circulating tumor cells.

[0147] It should be noted that, for circulating tumor cells screened out from sample cells, the screened circulating tumor cells are transmitted to the circulating tumor cell capture module by using the data transmission unit, so that the latter can capture the circulating tumor cells.

[0148] Furthermore, the circulating tumor cell capture module is a high-precision directional capture of screened circulating tumor cells, based on magnetic field manipulation technology and multi-channel single-cell confinement microstructure design, combined with a microfluidic system to achieve precise cell capture, including a circulating tumor cell positioning unit, a circulating tumor cell capture unit and a circulating tumor cell confinement unit.

[0149] Specifically, the circulating tumor cell positioning unit uses an external magnetic field to precisely operate the screened circulating tumor cells, guide the circulating tumor cells to a designated capture area, and attach magnetic particles to the surface of the circulating tumor cells through magnetic labeling technology. Under the action of , the circulating tumor cells move along the magnetic field lines, then,

[0150] in, represents the magnetic moment of magnetic particles attached to the surface of circulating tumor cells, express The magnetic field strength at the moment, represents the initial magnetic field strength, Indicates the control coefficient, which is used to adjust the sensitivity of magnetic field changes. It is set by the implementer according to the actual application scenario. represents the real-time displacement difference of circulating tumor cells, Represents the magnetic field force, acting on magnetic particles, used to guide the movement of circulating tumor cells.

[0151] It should be noted that when circulating tumor cells deviate from the predetermined route of the magnetic field, the sensor collects the real-time displacement difference of the cells and uses the collected real-time displacement difference to generate a signal. , dynamically adjust the magnetic field force

[0152] small and directed, thereby completing the precise guidance of circulating tumor cells.

[0153] Furthermore, the circulating tumor cell capture unit utilizes different flow rates and fluid environments in each channel of the multi-microfluidic channels, and controls the movement trajectories of different cells in different channels by adjusting the flow rate of each channel, and ensures that the circulating tumor cells enter the set capture area, as follows:

[0154] By controlling the flow rate in each channel, a differentiated fluid environment is generated, and the dynamic regulation of shear stress is achieved, then,

[0155] in, represents the viscosity of the fluid, represents the velocity difference of the flow velocity in the channel, represents the distance between fluid layers in the channel, represents dynamically regulated shear stress;

[0156] Monitoring the deformation of circulating tumor cells based on dynamically regulated shear stress combined with real-time imaging , according to the monitored deformation of circulating tumor cells, the directional capture of circulating tumor cells is achieved, then,

[0157] in, represents the shape of circulating tumor cells, represents the dynamically regulated shear stress, represents the elastic modulus of circulating tumor cells, represents the velocity coefficient of circulating tumor cells responding to shear stress, Indicates the time of shear stress action;

[0158] Different cell capture devices are designed at the end of each microfluidic channel. The deformation of circulating tumor cells in the channel is determined according to the shear stress in different channels. Different, by setting the standard threshold of deformation of circulating tumor cells under different shear stresses , then there is,

[0159] If the deformation of circulating tumor cells exceeds the standard threshold of deformation, the flow rate is reduced through a feedback mechanism, thereby reducing the shear stress;

[0160] If the deformation of circulating tumor cells is lower than the standard threshold of deformation, the flow rate is increased through a feedback mechanism, thereby increasing the shear stress;

[0161] If the shape variable of circulating tumor cells reaches the standard shape variable threshold, the current circulating tumor cells are guided into a specific area for targeted capture.

[0162] Furthermore, the circulating tumor cell confinement unit is a confinement microstructure designed based on microfluidic technology. The confinement microstructure is a microporous structure set in the circulating tumor cell capture area to limit a movement space for the cells to be captured. The microporous structure is designed based on geometric constraints, as follows:

[0163] in, is the surface tension coefficient between circulating tumor cells and microstructures, is the radius of the microstructure, is the radius of circulating tumor cells to be captured, The size of the geometric constraint force can be dynamically adjusted according to the radius of circulating tumor cells, thereby achieving accurate capture and confinement of circulating tumor cells.

[0164] It should be noted that for the directed capture of circulating tumor cells, the screened target cells are first positioned and guided by the circulating tumor cell positioning unit, and then the target cells are directionally captured using the circulating tumor cell capture unit. While capturing, the capture area level is spatially restricted using the circulating tumor cell confinement unit to achieve accurate capture of the target cells.

[0165] Furthermore, if the functions are implemented in the form of software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of the present invention, or the part that contributes to the prior art, or the part of the technical solution, can be embodied in the form of a software product. The computer software product is stored in a storage medium, including several instructions for enabling a computer device (which can be a personal computer, server, or network device, etc.) to perform all or part of the steps of the methods described in various embodiments of the present invention. The aforementioned storage medium includes: various media that can store program codes, such as a USB flash drive, a mobile hard disk, a read-only memory (ROM), a random access memory (RAM), a magnetic disk or an optical disk.

[0166] The logic and / or steps represented in the flowchart or otherwise described herein, for example, can be considered as an ordered list of executable instructions for implementing logical functions, and can be embodied in any computer-readable medium for use by an instruction execution system, device or apparatus (such as a computer-based system, a system including a processor, or other system that can fetch instructions from an instruction execution system, device or apparatus and execute instructions), or in conjunction with such instruction execution systems, devices or apparatuses. For the purposes of this specification, "computer-readable medium" can be any device that can contain, store, communicate, propagate or transmit a program for use by an instruction execution system, device or apparatus, or in conjunction with such instruction execution systems, devices or apparatuses.

[0167] More specific examples of computer-readable media (a non-exhaustive list) include the following: an electrical connection with one or more wires (electronic device), a portable computer disk case (magnetic device), a random access memory (RAM), a read-only memory (ROM), an erasable and programmable read-only memory (EPROM or flash memory), an optical fiber device, and a portable compact disk read-only memory (CDROM). In addition, the computer-readable medium may even be a paper or other suitable medium on which the program is printed, since the program may be obtained electronically, for example, by optically scanning the paper or other medium, followed by editing, deciphering or, if necessary, processing in another suitable manner, and then stored in a computer memory.

[0168] Additionally, in order to provide a concise description of exemplary embodiments, all features of an actual embodiment may not be described (ie, those features that are not relevant to the best mode presently contemplated for carrying out the invention or those features that are not relevant to implementing the invention).

[0169] It will be appreciated that in the development of any actual implementation, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but will be a routine task of design, fabrication, and production for those of ordinary skill having the benefit of this disclosure without undue experimentation.

[0170] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A microfluidic chip rapid detection system, comprising: The sample pre-processing and fluid control module is used to perform preliminary processing on the input sample cells, and is characterized by comprising a sample dilution and separation unit and a fluid drive and control unit, specifically: The sample blood is subjected to sample pretreatment, sample separation, and dilution treatment from its original state to form a sample fluid to be detected, and the sample fluid is made to flow into a microfluidic chip; And, the sensing detection and signal processing module is used to detect the sample cells and send the detected data signals to the circulating tumor cell capture module, including an optical sensor detection unit, a signal data processing unit and a data transmission unit, specifically: An optical sensor is integrated in the microfluidic channel, and a light source is used to illuminate the sample cells to generate discrete light signals and fluorescence signals. The circulating tumor cell features are extracted based on the generated discrete light signals and fluorescence signals, thereby detecting the circulating tumor cells, and sending the detection results to the circulating tumor cell capture module; The method of using a light source to illuminate sample cells to generate discrete light signals and fluorescence signals is specifically as follows: Discrete optical signal acquisition, in, represents the initial intensity of the incident light, represents the cell radius, represents the distance from the cell to the optical sensor, represents the influencing factor function, represents the scattering angle, represents the wavelength of light of the optical sensor, represents the refractive index of the cells in the sample cell, represents the acquired discrete optical signal; Fluorescence signal acquisition, in, represents the initial intensity of the incident light, represents the fluorescence quantum yield, represents the fluorescence absorbance, represents the concentration of fluorescent dye, represents the acquired fluorescence signal; The detection of circulating tumor cells is specifically as follows: According to the extracted circulating tumor cell characteristics, a circulating tumor cell scoring model is constructed, then, in, represents the forward scattering intensity, represents the side scattering intensity, represents the relative fluorescence intensity, represents the bias term, Respectively represent the weight coefficient of each feature; Screening of circulating tumor cells was performed based on the scoring results, as follows: The score of the sample cells under the circulating tumor cell scoring model is , the score of normal cells in the circulating tumor cell scoring model is , the score of circulating tumor cells under the circulating tumor cell scoring model is , according to the score, circulating tumor cells are screened, then, Normal cells and circulating tumor cells are screened according to the scoring results of sample cells under the circulating tumor cell scoring model; If the score of the sample cell in the circulating tumor cell scoring model satisfies the formula , indicating that the sample cells corresponding to the current score are a collection of circulating tumor cells and normal cells, and the current sample cells need to be screened a second time; The secondary screening is as follows: According to the characteristics of the constructed data, a dynamic screening mechanism is constructed. Scoring mechanism based on behavioral data characteristics, ； Scoring mechanism under spectral data, in, represents the movement speed of sample cells in the microfluidic channel, represents the velocity change rate of the sample cell in the microfluidic channel, represents the acceleration of the sample cell in the microfluidic channel, represents the fluorescence response at different wavelengths, Indicates the upper limit of wavelength types, Indicates the type index of wavelength, Indicates the degree of influence of spectral characteristics at different wavelengths on the score. Respectively represent the influence of speed, acceleration and speed change rate on the score. Indicates the scoring result under the behavior data characteristics, Indicates the scoring results under spectral data; Combining the scoring models under the two data characteristics to construct a comprehensive scoring model, we have: in, They represent the weight coefficients of behavioral scoring and spectral scoring respectively, which are set by the implementers according to the actual application scenarios. Indicates the scoring result under the behavior data characteristics, Indicates the scoring results under spectral data; Screening is performed based on behavioral scores, spectrum scores, and comprehensive score models, as follows: Using normal cells to set behavioral scoring thresholds , spectral scoring standard threshold , and the comprehensive scoring criteria threshold , Screening of circulating tumor cells in sample cells according to a set standard threshold; Also includes: The circulating tumor cell capture module is used to capture circulating tumor cells according to the data signal, including a circulating tumor cell positioning unit, a circulating tumor cell capture unit and a circulating tumor cell confinement unit, specifically: The screened circulating tumor cells are captured in a targeted manner based on magnetic field manipulation technology and multi-channel single-cell confined microstructure design, combined with a microfluidic system to achieve precise cell capture.

2. A microfluidic chip rapid detection system as claimed in claim 1, characterized in that: The sample preprocessing is specifically as follows: Set the input biological blood sample to , and including sample cells And impurities , then there is, The total number of sample cells is ; The blood sample is filtered using a membrane to remove impurities. in, represents the sample cells in the blood sample, Indicates impurities in blood samples, It indicates the proportion of sample cells in the blood sample after filtration, which is used to control the degree of cell filtration and satisfies the formula , Represents the proportion coefficient of sample cells; Update the data of the filtered sample cells, then we have, , Indicates impurities after filtration, Represents a blood sample after impurities have been filtered out.

3. A microfluidic chip rapid detection system as claimed in claim 2, characterized in that: The sample dilution is specifically implemented as follows: After the sample cells are separated, the total number of sample cells is The volume of the sample solution is , the sample cells are at the bottom of the centrifuge tube, and the concentration of the sample cells is Then, in, Represents the total number of sample cells, represents the volume of sample solution, Indicates the concentration of sample cells after separation; By adding the volume of buffer solution, the initial sample volume Dilution is performed to reduce the viscosity of the sample cells and ensure smooth flow of the fluid in the microfluidic chip. in, Represents the total number of sample cells, represents the volume of sample solution, represents the concentration of sample cells after adding buffer, represents the volume of buffer added; By adding buffer, the viscosity of the sample cells can be adjusted, then, in, represent the initial viscosity of the sample solution and the viscosity of the buffer solution, respectively. represents the volume of sample solution, represents the volume of buffer added, represents the viscosity of sample cells after adjustment; The dilution of the sample cells is controlled by monitoring the concentration and viscosity of the sample cells after adding the buffer, as follows: The objective function is set based on the sample cell concentration and viscosity after adding the buffer solution, and the objective function is set based on the volume of the buffer solution added. For optimization, we have in, Indicates that the sample cells meet the minimum concentration requirement for microfluidic chip detection. represents the maximum viscosity at which sample cells can flow into the microfluidic channel, Represents the objective function. By limiting the result of the objective function, the volume of the added buffer is optimized. Then, for The result satisfies the formula as well as , the buffer volume corresponding to the objective function at this time is the volume of buffer added.

4. A microfluidic chip rapid detection system as claimed in claim 3, characterized in that: The circulating tumor cell localization unit is specifically as follows: Through magnetic labeling technology, magnetic particles are attached to the surface of circulating tumor cells. Under the action of , circulating tumor cells move along the magnetic field lines, then, in, represents the magnetic moment of magnetic particles attached to the surface of circulating tumor cells, express The magnetic field strength at the moment, represents the initial magnetic field strength, Indicates the control coefficient, which is used to adjust the sensitivity of magnetic field changes. It is set by the implementer according to the actual application scenario. represents the real-time displacement difference of circulating tumor cells, Represents the magnetic field force, acting on magnetic particles, used to guide the movement of circulating tumor cells.

5. A microfluidic chip rapid detection system as claimed in claim 4, characterized in that: The specific method of directional capture of the screened circulating tumor cells is as follows: By controlling the flow rate in each channel, a differentiated fluid environment is generated, and the dynamic regulation of shear stress is achieved, then, in, represents the viscosity of the fluid, represents the velocity difference of the flow velocity in the channel, represents the distance between fluid layers in the channel, represents dynamically regulated shear stress; Monitoring the deformation of circulating tumor cells based on dynamically regulated shear stress combined with real-time imaging , according to the monitored deformation of circulating tumor cells, the directional capture of circulating tumor cells is achieved, then, in, represents the shape of circulating tumor cells, represents the dynamically regulated shear stress, represents the elastic modulus of circulating tumor cells, represents the velocity coefficient of circulating tumor cells responding to shear stress, Indicates the time of shear stress action; Different cell capture devices are designed at the end of each microfluidic channel. The deformation of circulating tumor cells in the channel is determined according to the shear stress in different channels. Different, by setting the standard threshold of deformation of circulating tumor cells under different shear stresses ,If the shape variable of circulating tumor cells reaches the standard shape variable threshold, the current circulating tumor cells are guided into a specific area for directional capture.

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