CN211603212U - Cell in-vivo capturing system to be detected - Google Patents

Abstract

The utility model discloses an in-vivo cell capturing system to be detected, which comprises an immune micro-nano bubble supply device, an acoustic resonance immune guide body in-vivo conveying device, an ultrasonic emission device and an acoustic resonance immune guide body; the immune micro-nano bubble supply device provides immune micro-nano bubbles coupled with antibodies, and the immune micro-nano bubbles are injected into a body and are specifically combined with cells to be detected to form a bubble complex; the in-vivo conveying device of the guide body conveys the acoustic resonance immune guide body to an in-vivo area to be detected; the ultrasonic transmitting device transmits ultrasonic waves to excite the acoustic resonance immune guide body in the body to generate a local strong field; the surface of the acoustic resonance immune guide body is coupled with an antibody capable of identifying the cell to be detected, the antibody is specifically combined with the cell to be detected when being conveyed into the body to capture the cell to be detected, and the local strong field is utilized to adsorb the bubble complex to capture the cell to be detected. The utility model discloses can improve the quantity of catching of internal trace cell that awaits measuring, promote the accuracy of follow-up detection.

Description

Cell in-vivo capturing system to be detected

Technical Field

The utility model belongs to the technical field of cell in-vivo detection, especially, relate to a cell system of catching in vivo that awaits measuring.

Background

In recent years, despite advances in cancer therapy, tumor metastasis remains a final challenge in human struggle with cancer therapy and is the leading culprit in cancer-related deaths. Circulating Tumor Cells (CTCs) are important mediators of tumorigenesis and metastasis, and during tumorigenesis and development, CTCs are released into blood circulation by tumors or metastasis foci at primary sites, so that the CTCs are colonized on tissues or organs far away from the body to form new metastasis foci. CTC as a safe and reliable biological marker has the advantages of no wound, easy acquisition, high repeatability and the like, is rapidly applied and developed in the field of malignant tumors, and related researches cover multiple fields of early diagnosis, treatment, prognosis evaluation, pathogenesis and the like of tumors. However, although the blood of tumor patients may contain hundreds of CTCs, a blood sample of 5-10 ml (accounting for only 0.1% of the total blood volume) in the traditional detection mode usually contains only a small amount of CTCs, and the tumor cells in this order are not enough for comprehensive detection of biological behaviors such as tumor molecular biological heterogeneity and drug resistance mutation. The low magnitude of CTC numbers can lead to false negative results in tumor blood samples, particularly in early tumor detection or tumor recurrence detection, leading to misleading clinical decision making, leading to irreparable loss of tumor patient treatment and survival prognosis.

Currently, CTC detection technologies can be largely divided into three major categories based on the physical and biological properties of CTCs, namely: a nucleic acid expression-based CTC detection format, a cell physical property-based CTC detection format, and a cell surface antibody-based CTC detection format.

The detection of CTC through nucleic acid expression mainly detects free nucleic acid in serum of a tumor patient according to the fact that CTC has DNA or RNA expression which is different from blood component cells, however, nucleic acid carried by tumor apoptosis cells, exosomes and other nucleated cells in peripheral blood can obviously influence the detection result, and the technology has obvious defects. At present, CTC has larger volume and density compared with blood component cells, the charge distribution on the surface of the cells is also different from that of other cells, and in addition, part of CTC can also show different cell activities and other characteristics, and the specific differences are gradually known and applied to the detection and screening of the cells. With the development of a large number of related researches on tumor cell membrane protein and related functional verification, a theoretical basis and a new choice are provided for the detection and sorting of CTC, so that the sensitivity and specificity of the CTC detection technology based on cell surface antibodies are obviously improved. The most widely used technology at present relies primarily on CTC cell surface specific antibodies.

Epithelial cell adhesion molecules (EpCAM) are widely used in cell surface antibody-based CTC detection because they are expressed in almost all cells of epithelial origin and not in blood component cells. Epithelial tumor cells are enriched and combined with specific anti-EpCAM antibody coupled immunomagnetic beads, and are screened by a specific magnetic field, so that the epithelial tumor cells are already used for detecting the CTC of malignant tumors such as lung cancer, breast cancer, colorectal cancer and the like. Two representative CTC detection technologies developed based on this principle are currently available, namely the commercial CellSearch system produced by Veridex, a product approved by the Food and Drug Administration (FDA) and the Chinese Food and Drug Administration (CFDA), and the only global sampling needle produced by gilipi, germany, which captures CTCs in vivo and obtains CFDA approval.

Although the CellSearch system is a more standard CTC detection method in the prior art and has already been applied to partial clinical trials, the disadvantage of this system is mainly its low sensitivity (only about 1 CTC cell per ml of blood is detected), and a single detection usually requires a large blood sample, and an efficient detection of CTCs can be achieved only in peripheral blood of patients with distant metastases from tumors, see: andree, K.C., van Dalum, G., Terstappen, L.W. Challenges in circulating tumor cell detection by the cell search system, MolOncol 2016; 10: 395-.

Compared with the CellSearch system, the CellColctor sampling needle related research proves that the cell quantity acquisition and sensitivity in CTC detection are more advantageous. However, the CellCollector sampling needle has limited ability to capture CTCs in vivo, has poor attraction and capture ability for CTC cells flowing in blood vessels, can only capture CTC cells flowing through the surface of the sampling needle, and cannot capture CTC cells far away from the sampling needle, see: vermesh, O.O., Aalipor, A.A., Ge, T.J., Saenz, Y.Guo, Y.Y., Alam, I.S., Park, S.M., Adelson, C.N., Mitsutake, Y.Y., Vilches-Moure, J.J., et al, An intravasular magnetic wire for the high-through put regenerative recovery of circulating cells in v.Nat Biomed Eng2018, 2: 696-.

SUMMERY OF THE UTILITY MODEL

In view of the deficiencies in the prior art, the utility model provides a cell system is caught in vivo to await measuring for improve the capture quantity of internal trace cell (for example circulating tumor cell CTC) that awaits measuring, promote the accuracy of follow-up detection.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

an in-vivo capturing system for cells to be detected comprises an immune micro-nano bubble supply device, an acoustic resonance immune guide body in-vivo conveying device, an ultrasonic emitting device and an acoustic resonance immune guide body; wherein the content of the first and second substances,

the immune micro-nano bubble supply device is used for providing immune micro-nano bubbles, antibodies capable of identifying cells to be detected are coupled to the surfaces of the immune micro-nano bubbles, and the immune micro-nano bubbles are injected into an in-vivo region to be detected and are specifically combined with the cells to be detected to form an immune micro-nano bubble complex;

the acoustic resonance immune guide body in-vivo conveying device is used for conveying the acoustic resonance immune guide body from the outside to the in-vivo area to be detected and from the in-vivo area to be detected to the outside;

the ultrasonic transmitting device is used for transmitting ultrasonic waves outside the body to excite the acoustic resonance immune guide body in the body to generate a local strong field;

the surface of the acoustic resonance immune guide body is coupled with an antibody capable of identifying cells to be detected, and the antibody is used for being specifically combined with the cells to be detected to directly capture the cells to be detected when the antibodies are conveyed to an area to be detected in vivo, and the immune micro-nano bubble complex is adsorbed by the local strong field to capture the cells to be detected.

The immune micro-nano bubble supply device is a micro-fluidic cavity device and comprises a primary cavity, a secondary cavity and a tertiary cavity which are sequentially connected; the primary cavity channel comprises a gas cavity channel, a liquid lipid cavity channel, a bubble forming cavity channel and an antibody conveying cavity channel; the secondary cavity channel is formed by coupling and assembling an antibody and micro-nano bubbles; the third-level cavity channel is used for screening and obtaining immune micro-nano bubbles with the surfaces coupled with antibodies capable of identifying cells to be detected.

Wherein the diameter of the immune micro-nano bubble is 1-10 μm.

Wherein the material of the acoustic resonance immune guide body is a flexible material; the transverse wave speed of the acoustic resonance immune guide body is smaller than the longitudinal wave speed of the body fluid of the region to be detected in the body.

Wherein, the material of the acoustic resonance immune guide body is a biomedical high polymer material.

Wherein the acoustic resonance immune guide body is a hollow or solid linear guide wire; the cross section of the acoustic resonance immune guide body is triangular, circular or rectangular.

Wherein, the surface of the acoustic resonance immune guide body is coated with a hydrophilic or hydrophobic coating, and an antibody which can identify the cells to be detected is coupled and assembled on the coating.

The circumference diameter of the acoustic resonance immune guide body is 50-500 mu m, and the working frequency of the local strong field generating mode is 1-10 MHz.

The acoustic resonance immune guide body internal conveying device is a puncture outer sheath tube, a pre-installed puncture needle is used for puncturing the internal region to be detected through the puncture outer sheath tube, and the acoustic resonance immune guide body is conveyed to the internal region to be detected after internal puncture through the puncture outer sheath tube.

The ultrasonic transmitting device comprises a signal generator, a power amplifier and an ultrasonic transducer, wherein a signal generated by the signal generator is amplified by the power amplifier and then excites the ultrasonic transducer to transmit ultrasonic waves.

The in-vivo cell capturing system for the cells to be detected further comprises a microfluidic cell sorting detection device, and the microfluidic cell sorting detection device is used for detecting the cells to be detected captured by the acoustic resonance immune guide body.

The cell to be detected is a circulating tumor cell, the antibody capable of identifying the cell to be detected is an antibody capable of identifying a tumor specific antigen, and the region to be detected in vivo is a blood vessel.

The embodiment of the utility model provides a cell that awaits measuring is at body capture system, send into the internal region of waiting to detect through the immune micro-nano bubble and the acoustic resonance immune guide body that have the antibody that can discern the cell that awaits measuring with the surface coupling, the acoustic resonance immune guide body can take place the specificity with the cell that awaits measuring on the one hand and combine direct capture cell that awaits measuring, on the other hand is under ultrasonic excitation, utilize the immune micro-nano bubble after "secondary sound source" absorption enrichment and the cell that awaits measuring that the acoustic resonance immune guide body produced local sound field, further catch the cell that awaits measuring of more quantity, the capture quantity of internal micro cell that awaits measuring (for example circulation tumor cell CTC) has been improved, promote the accuracy of.

Taking the capture of CTC as an example, compared with the CellSearch system in the prior art, the utility model can continuously collect and capture CTC in the process of continuous blood circulation in vivo, thereby continuously increasing the local concentration of CTC cells and capturing more CTC cells. Compare with the CellSearch system among the prior art, CellCollector sampling needle compares, the utility model discloses not only can catch the CTC cell of flowing through acoustic resonance immunity guide body surface, can also catch the CTC cell in the certain distance around the resonance immunity guide body through the adsorption affinity of local strong field, can catch more quantity of CTC cell.

In addition, the acoustic manipulation mainly utilizes the functions of capturing, arranging, moving, rotating, releasing and the like of the particles under the action of acoustic radiation force, drag force or shearing force caused by acoustic flow and the like applied to the particles in an acoustic field. Because supersound has been widely used in clinical imaging detection to it has been verified that low-intensity supersound can not cause obvious damage to the organism, therefore the particle is controlled to the supersound is a non-contact, noninvasive, convenient method of controlling, catches CTC in-process through the immune micro-nano bubble of sound manipulation can not cause cell damage and ensured the biological activity of CTC, promptly, the utility model provides a cell in vivo that awaits measuring catches system and working method thereof is safe and reliable more.

Drawings

FIG. 1 is a schematic structural diagram of an in vivo capture system for a test cell according to an embodiment of the present invention;

fig. 2 is a diagram of the embodiment of the present invention in which an antibody capable of identifying a cell to be detected is coupled to the immune micro-nano bubble;

fig. 3 is a diagram of an immune micro-nano bubble complex combined by immune micro-nano bubbles and cells to be detected in the embodiment of the present invention;

fig. 4 is a schematic structural diagram of an immune micro-nano bubble supply device in an embodiment of the present invention;

figure 5 is an exemplary illustration of the structure of an acoustic resonance immune guide in vivo delivery device and its process of delivering an acoustic resonance immune guide in an embodiment of the present invention;

fig. 6 is a schematic structural diagram of an ultrasonic transmitter in an embodiment of the present invention;

fig. 7 is a schematic structural view of an acoustic resonance immune guide according to an embodiment of the present invention;

fig. 8 is a schematic structural diagram of a microfluidic cell sorting and detecting device according to an embodiment of the present invention;

FIG. 9 is a flowchart illustrating an in vivo cell trapping method according to an embodiment of the present invention;

fig. 10 to 12 are force distribution diagrams of immune micro-nano bubbles with different sizes and diameters around an acoustic resonance guide body with a diameter of 300 microns and a circular cross section.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention clearer, the following detailed description of the embodiments of the present invention will be described with reference to the accompanying drawings. Examples of these preferred embodiments are illustrated in the accompanying drawings. The embodiments of the invention shown in the drawings and described in accordance with the same are merely exemplary and the invention is not limited to these embodiments.

It should also be noted that, in order to avoid obscuring the invention with unnecessary details, only the structures and/or process steps that are closely related to the solution according to the invention are shown in the drawings, while other details that are not relevant to the invention are omitted.

Fig. 1 is a schematic structural diagram of an in vivo capture system for a test cell according to an embodiment of the present invention, as shown in fig. 1, the in vivo capture system for a test cell includes: the device comprises an immune micro-nano bubble supply device 10, an acoustic resonance immune guide body in-vivo conveying device 20, an ultrasonic emission device 30 and an acoustic resonance guide body 40. Preferably, the capture system of this embodiment further comprises a microfluidic cell sorting detection device 50.

The immune micro-nano bubble supply device 10 is used for supplying immune micro-nano bubbles. Referring to fig. 2 and 3, an antibody 2 capable of identifying a cell to be detected is coupled to the surface of the immune micro-nano bubble 1, and the immune micro-nano bubble 1 is injected into a region to be detected in vivo and specifically binds with the cell to be detected 3 to form an immune micro-nano bubble complex. In a specific embodiment, the test cell 3 is, for example, a Circulating Tumor Cell (CTC), the antibody 2 capable of recognizing the test cell is an antibody recognizing a tumor-specific antigen, and the in vivo test region is a blood vessel.

Specifically, the immune micro-nano bubble supply device 10 is a microfluidic channel device, and as shown in fig. 4, the microfluidic channel device includes a primary channel 11, a secondary channel 12, and a tertiary channel 13, which are connected in sequence.

Referring to fig. 4, the primary channel 11 includes a gas channel 11a, a liquid lipid channel 11b, a bubble-forming channel 11c, and an antibody delivery channel 11 d. The gas cavity 11a is used for supplying inert gas G, such as perfluoropropane, for forming the immune micro-nano bubble inner core; the liquid lipid channel 11b is used for supplying a liquid lipid material L, such as phospholipid, for forming an immune micro-nano bubble shell; the bubble forming cavity channel 11c is connected with the output ends of the gas cavity channel 11a and the liquid lipid cavity channel 11b, and the bubble forming cavity channel 11c is a cavity channel for forming the immune micro-nano bubble 1 by combining inert gas G and a liquid lipid material L; the antibody delivery channel 11d is used to supply an antibody 2 capable of recognizing a test cell. The secondary cavity channel 12 is connected with the output ends of the foam forming cavity channel 11c and the antibody conveying cavity channel 11d, the secondary cavity channel 12 is a cavity channel formed by coupling and assembling the antibody 2 and the immune micro-nano bubbles 1, an ultrasonic device 14 is arranged outside the cavity channel at the tail end of the secondary cavity channel 12, and the immune micro-nano bubbles assembled with the antibody are sorted by emitting ultrasonic waves 15. The three-stage cavity channel 13 is connected with the output end of the second-stage cavity channel 12, the three-stage cavity channel 13 is a cavity channel for screening and obtaining immune micro-nano bubbles of which the surfaces are coupled with antibodies capable of identifying cells to be detected, and the three-stage cavity channel 13 is used for screening and removing the immune micro-nano bubbles 1 with uniform particle sizes. Further, the microfluidic channel device further comprises a drainage channel 16, and the drainage channel 16 is used for discharging antibody which cannot be coupled and assembled, micro-nano bubbles, immune micro-nano bubbles coupled with the antibody and the like, wherein the particle size of the immune micro-nano bubbles is not in accordance with the requirement.

Compared with the traditional micro-nano bubble production technology, through the utility model discloses a microbubble that flow control chamber says that device produced can realize that the particle diameter is even, single to greatly optimize the internal efficiency of catching of micro-nano bubble.

The embodiment of the utility model provides an immune micro-nano bubble is the immune micro-nano bubble of lipid, and it is including coupling have the lipid shell and the inert gas inner core of the antibody that can discern the cell that awaits measuring, and its diameter is preferred in the scope of 1 mu m ~10 mu m.

In a preferred scheme, the immune micro-nano bubbles can be chemically or biologically modified to increase the adhesion efficiency and targeting property.

The acoustic resonance immune guide in-vivo conveying device 20 is used for conveying the acoustic resonance immune guide 40 from the outside to the in-vivo area to be tested, or from the in-vivo area to be tested to the outside.

In this embodiment, referring to fig. 5, the acoustic resonance immune guide in vivo delivery device 20 is specifically a puncture sheath 4 equipped with a puncture needle 5. The puncture sheath 4 comprises a puncture needle port 4a, a guide body port 4b and a puncture port 4 c. The process of conveying the acoustic resonance immune guide body 40 into the body by the acoustic resonance immune guide body in-vivo conveying device 20 specifically comprises the following steps: first, the puncture needle 5 is inserted into the puncture sheath 4 from the puncture needle port 4a, the puncture port 4c is punctured in order to align with a target region (for example, a blood vessel), the puncture needle 5 is withdrawn after the puncture is successful, and at this time, the acoustic resonance immune guide 40 is inserted into the puncture sheath 4 from the guide body port 4b and is sent to the punctured target region through the puncture port 4 c.

The ultrasonic emission device 30 is used for emitting ultrasonic wave outside the body to excite the acoustic resonance immune guidance body 40 in the body to generate a local strong field.

In this embodiment, referring to fig. 6, the ultrasonic transmitting device 30 includes a signal generator 31, a power amplifier 32, and an ultrasonic transducer 33, and after a signal generated by the signal generator 31 is amplified by the power amplifier 32, the ultrasonic transducer 33 is excited to transmit an ultrasonic wave.

Specifically, the ultrasonic transducer 33 may be one of a single-element ultrasonic transducer, a phased array ultrasonic transducer, a linear array ultrasonic transducer, and a convex array ultrasonic transducer; the resonant frequency of the acoustic resonance immune guide 40 determines, among other things, the drive frequency at which the ultrasound is emitted, and thus the center frequency of the ultrasound transducer 33. The signal generated by the signal generator 31 may be a continuous sinusoidal signal or a pulsed sinusoidal signal. In one embodiment, the signal generator 31 may be a programmable signal generator (AFG 3021, Tectronix), the power amplifier may be a 50dB linear power amplifier (325 LA, ENI), and the signal generator 31 generates a sinusoidal continuous signal that drives the ultrasound transducer 33 to generate ultrasound waves via the power amplifier 32.

Referring to fig. 7, an antibody 41 capable of recognizing a test cell is coupled to the surface of the acoustic resonance immune guide 40, and an antibody 41 capable of recognizing a test cell is coupled to at least a portion fed to the target region; further, in this embodiment, the surface of the acoustic resonance immune guide 40 is first coated with a hydrophilic or hydrophobic coating 44, and an antibody 41 capable of recognizing a cell to be detected is coupled and assembled on the coating 44. The acoustic resonance immune guide body 40 is also provided with an insertion mark point 42 and an exit mark point 43. The acoustic resonance immune guide body 40 is used for being specifically combined with cells to be detected to directly capture the cells to be detected when being conveyed to an in-vivo region to be detected, and adsorbing the immune micro-nano bubble complex by using the local strong field to capture the cells to be detected.

Referring to fig. 5 and 7, when the acoustic resonance immune guide 40 is placed into the puncture sheath 4 from the guide port 4b, the placement index point 42 of the acoustic resonance immune guide 40 is brought flush with the guide port 4 b. After the acoustic resonance immune guide 40 finishes capturing the cells to be tested, the acoustic resonance immune guide 40 is slowly withdrawn from the guide port 4b to the withdrawal marking point 43 which is flush with the guide port 4b, and then the whole puncture sheath tube 4 is withdrawn from the target area.

The material of the acoustic resonance immune guide body is a flexible material and is a biomedical high polymer material, such as polylactic acid-glycolic acid copolymer, polyvinyl chloride, polyethylene, polytetrafluoroethylene, polyurethane and the like.

The transverse wave velocity of the acoustic resonance immune guide body is smaller than the longitudinal wave velocity of body fluid (such as blood in blood vessels) of a region to be detected in the body. The acoustic resonance guide body is a hollow or solid linear guide wire. The cross section of the acoustic resonance guide body is triangular, circular or rectangular.

In a preferable scheme, the circumference diameter of the acoustic resonance immune guide body is 50-500 μm, and the working frequency of the local strong field generating mode is 1-10 MHz.

In a preferred embodiment, the material of the acoustic resonance immune guide is poly (lactic-co-glycolic acid), PLGA, with longitudinal wave speed of 2114m/s and transverse wave speed of 532 m/s.

In a preferred embodiment, the acoustic resonance immune guide may be chemically or biologically modified to increase its adhesion efficiency and targeting. For example, the surface of the acoustic resonance immune guide is coated with a hydrophilic or hydrophobic coating, as desired.

In the embodiment of the present invention, the capturing system further includes a micro-fluidic cell sorting and detecting device 50, the micro-fluidic cell sorting and detecting device 50 is used for counting and detecting the cells to be detected captured by the acoustic resonance guiding body 40.

Specifically, the utility model provides a detection device is selected separately to micro-fluidic cell 50 is based on the micro-fluidic cell of supersound. As shown in fig. 8, the microfluidic cell sorting detection apparatus 50 includes: a cell to be detected fluorescent staining pool 51, a single cell flow channel 52, an ultrasonic device 53 and a fluorescent microscope imaging system 54. The cell to be detected fluorescent staining cell 51 is used for carrying out fluorescent staining on the cell to be detected captured by the in vivo capturing system, thereby realizing the visibility of the cell to be detected. The single cell flow channel 52 is connected with the cell fluorescent staining pool 51 to be detected, the ultrasonic device 53 is arranged outside the channel of the single cell flow channel 52, and an ultrasonic focusing wave emitted by the ultrasonic device 53 forms a sound field, so that a cell group in the cell to be detected subjected to fluorescent staining flows through the single cell flow channel 52 and realizes the regular arrangement of single cells, thereby being beneficial to the detection of a downstream fluorescent microscope imaging system 54. The fluorescence microscope imaging system 54 is disposed at the end of the single-cell flow channel 52, and is used for identifying, counting and detecting the cells to be detected which flow through the single-cell flow channel 52 and are labeled by fluorescence.

Referring to fig. 9, the method for operating the system for capturing test cells in vivo as described above is described, or a method for capturing test cells in vivo is provided, which includes the steps of:

s10, controlling the immune micro-nano bubble supply device to prepare immune micro-nano bubbles with the surfaces coupled with antibodies capable of identifying cells to be detected, and injecting the immune lipid micro-nano bubbles into an in-vivo region to be detected. In the step, after the immune micro-nano bubbles are injected into a region to be detected in a human body, the immune micro-nano bubbles and cells to be detected are specifically combined to form an immune micro-nano bubble complex.

S20, providing the acoustic resonance immune guide body and determining the working frequency of the acoustic resonance immune guide body. Specifically, according to the diameter and material parameters of the acoustic resonance immune guide body, the ultrasonic working frequency of the local strong field mode generated on the surface of the acoustic resonance immune guide body is theoretically predicted and experimentally measured. In practice, the acoustic resonance immune guide body can be placed in water, and the resonant frequency can be obtained by measuring the transmission spectrum to determine the working frequency. In a specific embodiment, the PLGA acoustic resonance guide has a diameter of 300 microns and is theoretically predicted to produce a local high field mode at an operating frequency of 1.5 MHz.

S30, controlling the acoustic resonance immune guide body internal conveying device to convey the acoustic resonance immune guide body with the surface coupled with the antibody capable of identifying the cell to be detected to the internal area to be detected. In the step, after the acoustic resonance immune guide body is conveyed to a region to be detected in a body, the acoustic resonance immune guide body and cells to be detected are subjected to specific binding to directly capture the cells to be detected.

S40, controlling an ultrasonic emission device to emit ultrasonic waves, exciting the acoustic resonance immune guide body to generate a local strong field, and enabling the acoustic resonance immune guide body to adsorb the immune micro-nano bubble complex so as to capture cells to be detected.

In the step, firstly, the micro-nano bubbles to be immunized and the acoustic resonance immune guide body are sent to the region to be detected in the body for a certain time, and then the ultrasonic transmitting device is controlled to transmit ultrasonic waves, wherein the frequency range of the ultrasonic waves transmitted by the ultrasonic transmitting device is determined according to the working frequency of the acoustic resonance guide body determined in the step S20, so that the requirement of resonance is met.

S50, controlling the acoustic resonance immune guide body in-vivo conveying device to convey the acoustic resonance guide body after the cells to be detected are captured to the outside of the body.

And S60, controlling the microfluidic cell sorting detection device to count and detect the cells to be detected captured by the acoustic resonance guide body.

The embodiment of the utility model provides a cell system and method of catching in vivo that awaits measuring has already verified its feasibility through numerical simulation.

Fig. 10 to 12 are force distribution diagrams of immune micro-nano bubbles with different sizes and diameters around a PLGA acoustic resonance guide body with a diameter of 300 microns and a circular cross section. Wherein, the arrows indicate the force direction, the color indicates the force intensity, the diameter of the immune micro-nano bubble in fig. 10 is 2 micrometers, the diameter of the immune micro-nano bubble in fig. 11 is 4 micrometers, and the diameter of the immune micro-nano bubble in fig. 12 is 6.6 micrometers. As can be seen from fig. 10 to 12, under the action of the external sound field, the local sound field generated by the acoustic resonance guide body through resonance can make the immune micro-nano bubbles subjected to the action of attraction force around the guide body and gather around the guide body, but the particle size of the immune micro-nano bubbles can influence the spatial distribution of the attraction force, so that the bubbles are attracted or repelled. Therefore, the immune micro-nano bubbles with proper particle size are required to be prepared by a microfluidic method, so that the microbubbles are captured and attracted as much as possible, and the diameter of the immune micro-nano bubbles is preferably in the range of 1-10 microns.

To sum up, the embodiment of the utility model provides a cell that awaits measuring is at body capture system and method, send into the internal region of waiting to detect through the immune micro-nano bubble and the acoustic resonance immune guide body that have the antibody that can discern the cell that awaits measuring with the surface coupling, the acoustic resonance immune guide body can take place the specificity with the cell that awaits measuring and combine directly to catch the cell that awaits measuring on the one hand, on the other hand is under ultrasonic excitation, utilize the immune guide silk of acoustic resonance as the immune micro-nano bubble of "secondary sound source" adsorption enrichment after combining with the cell that awaits measuring of producing local sound field, further catch the cell that awaits measuring of more quantity, the capture quantity of internal trace cell that awaits measuring (for example circulating tumor cell CTC) has been.

The foregoing is directed to embodiments of the present application and it is noted that numerous modifications and adaptations may be made by those skilled in the art without departing from the principles of the present application and are intended to be within the scope of the present application.

Claims (12)

1. An in-vivo cell capturing system to be detected is characterized by comprising an immune micro-nano bubble supply device, an acoustic resonance immune guide body in-vivo conveying device, an ultrasonic emitting device and an acoustic resonance immune guide body; wherein the content of the first and second substances,

the immune micro-nano bubble supply device is used for providing immune micro-nano bubbles, antibodies capable of identifying cells to be detected are coupled to the surfaces of the immune micro-nano bubbles, and the immune micro-nano bubbles are injected into an in-vivo region to be detected and are specifically combined with the cells to be detected to form an immune micro-nano bubble complex;

the acoustic resonance immune guide body in-vivo conveying device is used for conveying the acoustic resonance immune guide body from the outside to the in-vivo area to be detected and from the in-vivo area to be detected to the outside;

the ultrasonic transmitting device is used for transmitting ultrasonic waves outside the body to excite the acoustic resonance immune guide body in the body to generate a local strong field;

the surface of the acoustic resonance immune guide body is coupled with an antibody capable of identifying cells to be detected, and the antibody is used for being specifically combined with the cells to be detected to directly capture the cells to be detected when the antibodies are conveyed to an area to be detected in vivo, and the immune micro-nano bubble complex is adsorbed by the local strong field to capture the cells to be detected.

2. The in vivo capture system for cells to be tested according to claim 1, wherein the immune micro-nano bubble supply device is a microfluidic channel device comprising a primary channel, a secondary channel and a tertiary channel which are connected in sequence; the primary cavity channel comprises a gas cavity channel, a liquid cavity channel, a bubble forming cavity channel and an antibody conveying cavity channel; the secondary cavity channel is formed by coupling and assembling an antibody and micro-nano bubbles; the third-level cavity channel is used for screening and obtaining immune micro-nano bubbles with the surfaces coupled with antibodies capable of identifying cells to be detected.

3. The in vivo capture system for cells to be tested according to claim 2, wherein the diameter of the immune micro-nano bubbles is 1 μm to 10 μm.

4. The system for capturing cells to be tested according to claim 1, wherein the acoustic resonance immune guidance body is a guidance body made of a flexible material, and the transverse wave velocity of the acoustic resonance immune guidance body is smaller than the longitudinal wave velocity of the body fluid in the region to be tested in vivo.

5. The system for capturing cells to be detected according to claim 4, wherein the acoustic resonance immune guide is a guide made of biomedical polymer material.

6. The system for capturing test cells in vivo according to claim 4, wherein the acoustic resonance immune guide body is a hollow or solid linear guide wire, and the cross section of the acoustic resonance immune guide body is triangular, circular or rectangular.

7. The system for capturing test cells in vivo according to any one of claims 4 to 6, wherein the surface of the acoustic resonance immune guide is coated with a hydrophilic or hydrophobic coating, and an antibody capable of recognizing the test cells is coupled and assembled on the coating.

8. The system for capturing cells to be tested according to claim 4, wherein the circumference diameter of the acoustic resonance immune guide body is 50 μm to 500 μm, and the working frequency of the local strong field generation mode is 1MHz to 10 MHz.

9. The system for capturing test cells in vivo according to claim 1, wherein the acoustic resonance immune guide body in vivo delivery device is a puncture sheath, a pre-installed puncture needle is used to puncture the in vivo test region through the puncture sheath, and the acoustic resonance immune guide body is delivered to the in vivo test region after internal puncture through the puncture sheath.

10. The system for in vivo capturing of cells to be tested according to claim 1, wherein the ultrasonic emitting device comprises a signal generator, a power amplifier and an ultrasonic transducer, and a signal generated by the signal generator is amplified by the power amplifier and then excites the ultrasonic transducer to emit ultrasonic waves.

11. The system for in vivo capture of test cells according to claim 1, wherein the system for in vivo capture of test cells further comprises a microfluidic cell sorting detection device for detecting test cells captured by the acoustic resonance immune guide.

12. The system for in vivo capturing of test cells according to claim 1, wherein the test cells are circulating tumor cells, the antibody capable of recognizing the test cells is an antibody recognizing a tumor-specific antigen, and the in vivo test region is a blood vessel.

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