CN112718030A - Blood separation device based on acoustic fluid mechanics - Google Patents
Blood separation device based on acoustic fluid mechanics Download PDFInfo
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Abstract
The application relates to the technical field of biomedicine, and provides a blood separation device based on acoustic fluid mechanics. The blood separation device comprises a microfluidic chip, a microfluidic pump and a sound wave driving system, wherein the microfluidic pump is connected with a microchannel of the microfluidic chip through a hose, micropores arranged in an array manner are arranged on the bottom surface of the microchannel, and a piezoelectric transducer of the sound wave driving system is fixed on a glass substrate of the microfluidic chip. The micro fluid enters the micro flow channel under the action of the micro flow pump, particles in the micro fluid are separated according to different sound flow field forces of the particles with different particle sizes in a sound flow field, the particles with relatively large particle sizes are captured by the micropores by controlling the sound flow capturing force and the external flow field thrust force of the particles, and the particles with relatively small particle sizes move along with the flow field. The blood separation device not only has no damage to particles in blood and strong controllability, but also can simply and efficiently separate target bacteria from blood.
Description
Technical Field
The application relates to the technical field of biomedicine, in particular to a blood separation device based on acoustic fluid mechanics.
Background
Bacterial infection often causes various diseases, and the disease rate is high and the critical illness state is often caused, so that timely diagnosis and treatment are needed. At present, most of the research on the source of bacterial diseases requires the separation of specific bacteria from a mixture of cells and bacteria (such as blood) and then the analysis of the specific bacteria, so that the bacteria separation technology has great significance in the fields of clinical medicine, bioengineering and the like.
The traditional bacteria separation technology is to culture and separate in an agar culture medium, which consumes a lot of time and requires high technical level of operators. At present, a centrifugal technology is often adopted to separate a sample, however, the sample is often broken due to overlarge centrifugal force in the operation process, so that cells are inactivated; during separation, the equipment is required to be frequently started and closed to observe the sample, and the progress of sample separation is known, so that the operation burden of the equipment is increased, and the service life of the equipment is shortened; in addition, the centrifugal technology needs to manually take the test tube for observation, so that the operation is complicated, the risk of sample pollution is increased, and the separation effect of the sample is influenced; furthermore, the separation accuracy of the centrifugal technique is low.
In addition, there is a separation method based on the optical tweezers technique, but a large amount of heat is generated by long-time laser irradiation, thereby damaging the activity of the sample; the method based on the magnetic field technique requires the object to be operated to have magnetism; and the two methods have complex systems and high cost, so the two methods have low practicability.
In clinical blood samples, blood has high viscosity and complex components, but analysis results are urgently needed, so how to simply and efficiently separate target bacteria from blood on the premise of not damaging the structure of a sample is a technical problem to be solved at present.
Disclosure of Invention
The application aims at providing a blood separator based on acoustic fluid mechanics, blood separator not only is not damaged, the controllability is strong to the particle in the blood, and can separate out target bacterium from blood simply high-efficiently.
In order to achieve the above object, the present application provides a blood separation device based on acoustic hydrodynamics, comprising: the device comprises a micro-fluidic chip, a micro-fluidic pump and a sound wave driving system.
The micro-fluidic chip comprises a micro-channel and a glass substrate, wherein the micro-channel is bonded on the glass substrate.
The micro-channel comprises an inlet end, a micro-channel chamber and an outlet end, and the bottom surface of the micro-channel chamber is provided with micropores arranged in an array manner.
The micro-flow pump is connected with the inlet end and is used for controlling the flow rate of micro-fluid in the micro-flow channel and forming flow field thrust.
The sound wave driving system comprises a piezoelectric transducer, wherein the piezoelectric transducer is fixed on the surface of the glass substrate and used for driving the micro-holes arranged in the array to vibrate to generate sound flow capturing force.
And the microfluid enters the micro-channel through the inlet end under the action of the micro-fluid pump, and micro-particles with large particle sizes in the microfluid are captured by the micro-pores arranged in an array.
Preferably, the micro flow channel chamber is rectangular, and has a length of 10mm, a width of 1mm and a depth of 0.2 mm.
Preferably, the diameter of the micropores is 100 μm, and the depth is 80 μm; the distance between the adjacent micropores is 100 μm.
Preferably, the edges and corners of the micro-channel chamber are rounded.
Preferably, the piezoelectric transducer is fixed on the surface of the glass substrate through epoxy resin and is tightly attached to the micro flow channel.
Preferably, the micro flow channel is made of polydimethylsiloxane and is integrally formed by using a mold.
Preferably, the inlet end comprises an inlet channel, one end of the inlet channel is connected to one end of the micro flow channel chamber, the other end of the inlet channel is provided with a round hole-shaped inlet pipe interface, and the inlet pipe interface is connected with the micro flow pump; the outlet end comprises an outlet channel, one end of the outlet channel is connected to the other end of the micro-channel cavity, and a round-hole-shaped outlet pipe interface is formed at the other end of the outlet channel.
Preferably, the width of the inlet channel and the width of the outlet channel are set to be 200 μm, and the depth of the inlet channel and the outlet channel is consistent with the depth of the micro-channel chamber; the diameters of the inlet pipe connector and the outlet pipe connector are both 1 mm.
Preferably, the sound wave driving system further comprises a signal generator connected to the piezoelectric transducer for generating a signal of a set waveform.
Preferably, the sound wave driving system further comprises a voltage amplifier connected between the signal generator and the piezoelectric transducer for amplifying the signal generated by the signal generator.
The particles with relatively large particle sizes are captured by controlling the size of the acoustic flow capturing force and the external flow field thrust force applied to the particles, and the particles with relatively small particle sizes move along with the flow field. The blood separation device not only has no damage to particles in blood and strong controllability, but also can simply and efficiently separate target bacteria from blood.
Drawings
In order to more clearly explain the technical solution of the present application, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious to those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
Fig. 1 is an application scenario diagram provided in an embodiment of the present application;
FIG. 2 is a schematic structural diagram of a blood separation device based on acoustic hydrodynamics according to an embodiment of the present application;
FIG. 3 is a schematic structural view of a micro flow channel according to an embodiment of the present invention;
FIG. 4 is a side view of an acoustic-hydrodynamic based blood separation device provided by an embodiment of the present application;
fig. 5 is a schematic separation diagram of a blood separation device based on acoustic hydrodynamics according to an embodiment of the present application.
In the figure: 1-microfluidic chip, 11-microchannel, 12-glass substrate, 111-inlet end, 1111-inlet channel, 1112-inlet tube interface, 112-microchannel chamber, 1121-micropore, 113-outlet end, 1131-outlet channel, 1132-outlet tube interface, 2-microfluidic pump, 3-sound wave driving system, 31-piezoelectric transducer, 32-voltage amplifier and 33-signal generator.
Detailed Description
The technical solutions in the embodiments of the present application will be fully and clearly described below with reference to the drawings in the embodiments of the present application. It is to be understood that the embodiments described are only a few embodiments of the present application and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
Referring to fig. 1, an application scenario diagram provided in the embodiment of the present application is shown. The embodiment of the application provides a blood separator based on acoustic fluid mechanics, includes: the device comprises a micro-fluidic chip 1, a micro-fluidic pump 2 and a sound wave driving system 3. In the actual use process, the microfluidic chip 1 is arranged on a microscope objective table, the microfluidic pump 2 is connected with the microfluidic chip 1 through a hose, and the piezoelectric transducer 31 of the sound wave driving system 3 is fixed on the microfluidic chip 1. The micro-flow pump 2 injects micro-fluid into the micro-fluidic chip 1 through a hose, particles in the micro-fluid are separated under the action of flow field thrust and acoustic flow capture force generated by the acoustic wave driving system 3, and the specific separation condition can be observed through a microscope.
Specifically, referring to fig. 2, the microfluidic chip 1 includes a microchannel 11 and a glass substrate 12, and the microchannel 11 is bonded to the glass substrate 12.
Referring to fig. 3, in the embodiment of the present application, the microchannel 11 includes an inlet end 111, a microchannel chamber 112, and an outlet end 113. The micro flow channel chamber 112 is rectangular, and cylindrical micropores 1121 are formed in an array arrangement on a bottom surface thereof. The present embodiment employs a transparent closed microchannel chamber 112 to facilitate observation of the movement state and separation of particles at any time.
Preferably, the inlet end 111 comprises an inlet channel 1111, one end of the inlet channel 1111 is connected to one end of the microchannel chamber 112, the other end of the inlet channel 1111 is configured with a round hole-shaped inlet pipe interface 1112, and the inlet pipe interface 1112 is connected with the microfluidic pump 2 for injecting the blood sample; the outlet end 113 includes an outlet channel 1131, one end of the outlet channel 1131 is connected to the other end of the micro channel chamber 112, and the other end of the outlet channel 1131 is configured with an outlet pipe interface 1132 in a circular hole shape, for sending the separated bacteria particles to a collection vessel.
In the embodiment of the present application, the microfluidic pump 2 is connected to the inlet end 111, and is configured to control a flow rate of the microfluidic in the microchannel 11 and form a flow field thrust.
Referring to fig. 1, fig. 2 and fig. 3, in the embodiment of the present application, the acoustic driving system 3 includes a piezoelectric transducer 31, a voltage amplifier 32 and a signal generator 33, which are connected in sequence, where the piezoelectric transducer 31 is fixed on the surface of the glass substrate 12 by epoxy resin, and is tightly attached to the micro channel 11, and is configured to drive the micropores 1121 arranged in an array to vibrate and generate an acoustic current capturing force; the signal generator 33 is used for generating a signal with a set waveform; the voltage amplifier 32 is used for amplifying the signal generated by the signal generator 33.
In the embodiment of the present application, the micro fluid enters the micro channel 11 through the inlet end 111 under the action of the micro flow pump 2, and under the action of the acoustic flow field force, the micro pores 1121 arranged in an array captures the particles with relatively large particle size in the micro fluid, and the particles with relatively small particle size flow out of the blood separation device through the outlet end 113 along with the micro fluid and enter the collection vessel.
In the embodiment of the present application, the design principle of the microfluidic chip 1 is specifically as follows: first, the piezoelectric transducer 31 converts an electric signal into a mechanical signal and generates vibration, driven by the signal generator 33 and the voltage amplifier 32. Since the piezoelectric transducer 31, the glass substrate 12 and the micro channel 11 are bonded together, the vibration signal can be finally transmitted to the micro holes 1121 arranged in the array on the bottom surface of the micro channel chamber 112. Referring to fig. 4, due to viscous dissipation, a pressure difference is generated around the micro-pores 1121, and two micro-acoustic flows with opposite directions are formed inside and outside the micro-pores 1121, which are referred to as an in-pore acoustic flow and an out-pore acoustic flow, and the micro-fluidic chip 1 has a capture and separation function by using the two acoustic flows.
By utilizing the motion characteristics of the acoustic flow outside the pores, particles located near micropores 1121 can exhibit a Stokes drag force Fd(generated by the acoustic streaming field, the principal force) and acoustic radiation force Frad(generated by the acoustic field, secondary forces) are trapped in micropores 1121; the particles can then be firmly held in the micropores 1121 by the acoustic flow within the pores.
Wherein:
Fd＝6πηR(<v2>-u)；
specifically, R represents a particle radius, UradWhich is indicative of the force potential of the acoustic radiation,
representing gradient operators, p1And upsilon1Respectively representing first-order sound pressure and sound velocity, upsilon2Representing the second order sound velocity, u representing the particle motion velocity, η representing the volume change viscosity coefficient, ρmDenotes the density, k, of the mediummDenotes the compressibility factor, p, of the mediumpDenotes the density, k, of the particlespRepresenting the compressibility factor of the particles.
Referring to fig. 5, when particles of different sizes move in the microchannel 11 at the same flow rate, the moving particles are mainly subjected to two kinds of forces, the capturing force of the acoustic flow and the pushing force exerted by the flow field. Wherein the capture force of the sound flow is mainly expressed as Stokes drag force FdAnd acoustic radiation force FradAnd, as can be seen from the above calculation formula, the stokes drag force FdAnd acoustic radiation force FradAll of them are closely related to the radius of the particles, i.e. the particles with large radius are subjected to larger forces. Thus, when the trapping force of the acoustic flow acting on the particles is greater than the thrust exerted by the flow field, the particles are trapped; on the contrary, the particles move forward along with the flow field, thereby realizing the separation operation of the particles with different sizes.
Preferably, in the present embodiment, in consideration of the size and manufacturing accuracy of the experimental sample, the rectangular micro flow channel chamber 112 has a length of 10mm, a width of 1mm, and a depth of 0.2 mm; meanwhile, the micro-holes 1121 arranged in an array designed on the inner bottom surface are cylindrical, the diameter of the micro-holes 1121 is 100 μm, the depth of the micro-holes 1121 is 80 μm, and the distance between the adjacent micro-holes 1121 is 100 μm.
Preferably, in the embodiment of the present invention, the corners and edges of the micro flow channel chamber 112 are rounded, mainly to avoid the influence of the sound flow generated by the right-angle structure on the subsequent experimental analysis.
Preferably, in the embodiment of the present invention, the micro flow channel 11 is made of polydimethylsiloxane, and is integrally formed by using a mold. Polydimethylsiloxane is one of ideal materials for manufacturing the microfluidic chip 1, and has the characteristics of high thermal stability, strong biocompatibility, good air permeability and light transmittance, and easiness in observation under a microscope.
Preferably, in this embodiment, the width of the inlet channel 1111 and the outlet channel 1131 is set to 200 μm, and the depth is consistent with the depth of the micro flow channel chamber 112, so as to facilitate the smooth movement of the fluid; the diameters of the inlet pipe interface 1112 and the outlet pipe interface 1132 are both 1 mm.
In clinical tests, blood samples are mainly blood cells and bacteria, wherein the blood cells are mainly red blood cells and white blood cells, the particle size of the red blood cells is 6-10 mu m, the particle size of the white blood cells is 7-20 mu m, and the particle size of most bacteria is less than 5 mu m.
Based on the above-mentioned principle,the separation principle of the blood separation device in the embodiment of the application is as follows: the micro-flow pump 2 injects the blood sample into the micro-flow channel 11 through the inlet pipe interface 1112 at a certain speed, and at this time, various components (mainly blood cells and bacteria) in the blood move towards the outlet direction under the thrust of the external flow field; when the sound field is turned on, the micropores 1121 arranged in the array in the microfluidic chip 1 vibrate along with the sound field, and eddy current is generated around the micropores, and at this time, blood cells and bacteria flowing through the vicinity of the micropores 1121 are subjected to a drag force F mainly comprising stokes besides the flow field thrust forcedAnd acoustic radiation force FradCapture force of the acoustic flow. Under the same flow velocity, the blood cells and the bacteria receive the same flow field thrust, but the particle size of the blood cells is larger than that of the bacteria, so the acoustic flow capture force received by the blood cells is larger than that received by the bacteria. By controlling the vibration intensity of the piezoelectric transducer 31, the acoustic flow capturing force acting on the blood cells is greater than the flow field thrust, and the flow field thrust is greater than the acoustic flow capturing force acting on the bacteria particles, so that the blood cells with relatively large particle size are captured and fixed in the micropores 1121, and the bacteria particles with relatively small particle size move forward along with the flow field and enter the collection vessel through the outlet pipe interface 1132, thereby realizing the operation of separating the bacteria particles from the blood.
Further, on the basis of separating the target bacterial particles, the bacterial particles can be further separated by the above principle, such as separating gram-positive bacteria in the bacterial particles. Since the size difference of gram-positive bacteria is not significant, gram-negative bacteria can be separated by using the blood separation apparatus described in the examples of the present application by increasing the size of gram-negative bacteria by binding them with microspheres (10 μm) modified with the GN6 aptamer before the separation operation. Among them, the GN6 aptamer was specific in that it only bound to gram-negative bacteria and not to gram-positive bacteria.
In the embodiments of the present application, the target bacteria are not limited to any kind of bacterial particles, and the bacteria can be separated by the blood separation apparatus according to the embodiments of the present application as long as the particle size of the bacterial particles is significantly different from the particle size of other particles in the blood.
Separation techniques based on acoustic fluid mechanics have shown great potential in bacterial separation applications, with several advantages over other commonly used bacterial separation approaches.
The method has the following advantages: at present, sound wave generation and driving equipment is mature, the device is relatively simple, the operation is easy, the control of sound wave frequency and amplitude is easy to realize, and long-time training is not needed for operators.
The advantages are as follows: the acoustic wave driving belongs to a non-contact operation mode, namely, the acoustic wave driving is not in direct contact with fluid, meanwhile, no special requirements are required on the properties of an operation object, such as geometry, electromagnetism, optics, chemistry and the like, the property of a medium cannot be changed in the driving process, and good biocompatibility is shown.
The advantages are three: the acoustic wave drive is used as a high-efficiency, high-energy, clean (no pollution to fluid) and low-cost means, a special acoustic structure is designed according to the inherent nonlinear characteristic of the acoustic wave drive, the complex control on microfluid and particles can be realized, and a new way is provided for realizing the movement of the particles in biological media with high viscosity and high ionic strength.
According to the embodiment of the application, the particles with different particle sizes are separated based on different acoustic flow field forces borne by the particles in an acoustic flow place, and the particles with relatively large particle sizes are captured by controlling the acoustic flow capturing force borne by the particles and the external flow field thrust, and the particles with relatively small particle sizes move along with the flow field. In addition, the separation of the blood cells and the bacteria in the blood can be realized, and the separation of gram-positive bacteria and gram-negative bacteria in the bacteria can be further realized. Meanwhile, the experimental device is simple, the cost is low, and the technical requirements on operators are not high.
According to the technical scheme, the blood separating device based on acoustic fluid mechanics is provided, the blood separating device is not only harmless to particles in blood, strong in controllability, but also capable of simply and efficiently separating target bacteria from blood.
The present application has been described in detail with reference to specific embodiments and illustrative examples, but the description is not intended to limit the application. Those skilled in the art will appreciate that various equivalent substitutions, modifications or improvements may be made to the presently disclosed embodiments and implementations thereof without departing from the spirit and scope of the present disclosure, and these fall within the scope of the present disclosure. The protection scope of this application is subject to the appended claims.
Claims (10)
1. A blood separation device based on acoustic hydrodynamics, comprising: the system comprises a micro-fluidic chip, a micro-fluidic pump and a sound wave driving system;
the micro-fluidic chip comprises a micro-channel and a glass substrate, and the micro-channel is bonded on the glass substrate; the micro-channel comprises an inlet end, a micro-channel chamber and an outlet end, and the bottom surface of the micro-channel chamber is provided with micropores arranged in an array manner;
the micro-flow pump is connected with the inlet end and is used for controlling the flow rate of micro-fluid in the micro-flow channel and forming flow field thrust; the sound wave driving system comprises a piezoelectric transducer, and the piezoelectric transducer is fixed on the surface of the glass substrate and is used for driving the micro-pores arranged in the array to vibrate to generate sound flow capturing force;
and the microfluid enters the micro-channel through the inlet end under the action of the micro-fluid pump, and micro-particles with large particle sizes in the microfluid are captured by the micro-pores arranged in an array.
2. The sonohydrodynamics-based blood separation device of claim 1 wherein the microchannel chamber is rectangular with a length of 10mm, a width of 1mm and a depth of 0.2 mm.
3. The acoustic-hydrodynamic based blood separation device of claim 1, wherein the micropores have a diameter of 100 μm and a depth of 80 μm; the distance between the adjacent micropores is 100 μm.
4. The device of claim 1, wherein the corners and edges of the microchannel chamber are rounded.
5. The device of claim 1, wherein the piezoelectric transducer is fixed on the surface of the glass substrate by epoxy resin and is tightly attached to the microchannel.
6. The device of claim 1, wherein the microchannel is made of polydimethylsiloxane and is integrally molded with a mold.
7. The sonohydrodynamics-based blood separation device of claim 1, wherein the inlet port comprises an inlet channel, one end of the inlet channel is connected to one end of the microchannel chamber, the other end of the inlet channel is configured with a round-hole-shaped inlet pipe interface, and the inlet pipe interface is connected with the microfluidic pump; the outlet end comprises an outlet channel, one end of the outlet channel is connected to the other end of the micro-channel cavity, and a round-hole-shaped outlet pipe interface is formed at the other end of the outlet channel.
8. The acoustiohydrodynamic based blood separation device of claim 7, wherein the width of the inlet channel and the outlet channel is set to 200 μm and the depth is the same as the depth of the microchannel chamber; the diameters of the inlet pipe connector and the outlet pipe connector are both 1 mm.
9. The acoustic-hydrodynamics-based blood separation device of claim 1 wherein the acoustic drive system further comprises a signal generator connected to the piezoelectric transducer for generating a signal of a set waveform.
10. The acoustic-hydrodynamics-based blood separation device of claim 9, wherein the acoustic drive system further comprises a voltage amplifier connected between the signal generator and the piezoelectric transducer for amplifying the signal generated by the signal generator.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113477283A (en) * | 2021-06-18 | 2021-10-08 | 电子科技大学长三角研究院(湖州) | Method for driving fluid to move by non-plasma metal photoinduced ultrasound and capturing device |
| CN114100707A (en) * | 2021-10-21 | 2022-03-01 | 天津大学 | Device for capturing micro-nano particles |
| CN114522649A (en) * | 2022-02-15 | 2022-05-24 | 浙江大学 | Acoustic particle capturing and track control method based on magnetofluid reconstruction |
-
2021
- 2021-01-25 CN CN202110095866.5A patent/CN112718030A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113477283A (en) * | 2021-06-18 | 2021-10-08 | 电子科技大学长三角研究院(湖州) | Method for driving fluid to move by non-plasma metal photoinduced ultrasound and capturing device |
| CN114100707A (en) * | 2021-10-21 | 2022-03-01 | 天津大学 | Device for capturing micro-nano particles |
| CN114522649A (en) * | 2022-02-15 | 2022-05-24 | 浙江大学 | Acoustic particle capturing and track control method based on magnetofluid reconstruction |
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