CN114100706A - Particle sorting method and system based on particle drift - Google Patents
Particle sorting method and system based on particle drift Download PDFInfo
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- CN114100706A CN114100706A CN202111212418.5A CN202111212418A CN114100706A CN 114100706 A CN114100706 A CN 114100706A CN 202111212418 A CN202111212418 A CN 202111212418A CN 114100706 A CN114100706 A CN 114100706A
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- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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Abstract
The invention discloses a particle sorting method and a system based on particle drift, which comprises the following steps: step 1: obtaining a calibration drift voltage of a target particle in the classified particles, comparing the calibration drift voltage with a preset drift voltage, if the calibration drift voltage is smaller than the preset drift voltage, executing the step 2, otherwise, executing the step 3; step 2: correcting the drift voltage until the calibrated drift voltage is greater than the preset drift voltage; and step 3: injecting the classified particles into the first microfluidic chip, and applying a drift voltage to the first microfluidic chip according to a preset drift voltage; and 4, step 4: and collecting the drifting particles after drifting, carrying out secondary sorting, and applying a preset drifting voltage to the first micro-fluidic chip to enable the particles needing to be separated, which are smaller than the target particle calibration drifting voltage in the classified particles, to be subjected to the external force of the preset drifting voltage and to be shifted in the first micro-fluidic chip, so as to realize the pretreatment of part of the particles needing to be separated.
Description
Technical Field
The invention belongs to the field of microfluidics, and particularly relates to a particle sorting method and system based on particle drift.
Background
The micro-fluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in the biological, chemical and medical analysis process into a micron-scale chip, and automatically completes the whole analysis process.
However, the existing inertial microfluidic technology utilizes the fluid inertia effect to induce the particles to migrate under the action of inertia force in the flow channel so as to realize accurate control, and has the advantages of simple flow channel structure, convenient operation, high control precision and the like.
Disclosure of Invention
The invention aims to provide a particle sorting method and system based on particle drift, which are used for solving the problems that the fluid inertia effect has strong dependence on the apparent size of particles and the precise control on particles with high concentration and similar sizes is difficult.
The purpose of the invention can be realized by the following technical scheme:
a particle sorting method based on particle drift comprises the following steps:
step 1: obtaining a calibration drift voltage of a target particle in the classified particles, comparing the calibration drift voltage with a preset drift voltage, if the calibration drift voltage is smaller than the preset drift voltage, executing the step 2, otherwise, executing the step 3;
step 2: correcting the drift voltage until the calibrated drift voltage is greater than the preset drift voltage;
and step 3: injecting the classified particles into the first microfluidic chip, and applying a drift voltage to the first microfluidic chip according to a preset drift voltage;
and 4, step 4: and collecting the drifting particles after drifting, and carrying out secondary sorting.
Further, the preset drift voltage is the maximum voltage borne by the first microfluidic chip.
Furthermore, the calibration drift voltage of the target particles is the voltage required by the target particles to generate the maximum drift under the action of the electric field.
Further, the step of injecting the classified particles into the first microfluidic chip and applying a drift voltage to the first microfluidic chip according to a preset drift voltage includes:
constructing a drift channel in the first microfluidic chip, wherein one end of the drift channel is a particle injection end, the other end of the drift channel is a collection end, and the collection end is connected with a collection pool;
and after the classified particles enter the drift channel in the first micro-fluidic chip through the injection end, applying a preset drift voltage to the first micro-fluidic chip, and after a preset time period, performing secondary separation on the drift particles in the collection pool.
Further, the obtaining of the calibration drift voltage of the target particles in the classified particles is specifically that the target particles are injected into the first microfluidic chip, the drift piezoelectric is loaded on the first microfluidic chip, whether the target particles exist in the collection pool is detected, until the target particles cannot be detected in the collection pool, the corresponding drift piezoelectric is the calibration drift voltage, and if the drift piezoelectric loaded on the first microfluidic chip reaches the maximum bearing voltage and the target particles are still detected in the collection pool, the maximum bearing voltage is the calibration drift voltage.
Further, the performing of the drift voltage correction specifically includes:
s1: extracting a preset drift voltage value and a calibrated drift voltage value;
s2: establishing a plane coordinate system and marking (a preset drift voltage value and a calibrated drift voltage value) as a first calculation point, and marking (a calibrated drift voltage value and a preset drift voltage value) as a second calculation point;
s3: calculating the distance between the first calculation point and the second calculation point, and marking as a correction number;
s4: and subtracting the correction number from the preset drift voltage value to finish the drift voltage correction.
Further, the secondary sorting comprises the following steps:
and injecting the solution containing the drifting particles in the collecting pool into the second microfluidic chip to complete secondary sorting.
Further, the drift channel is in a shape of a mosquito coil incense coil.
A particle sorting system based on particle drift, adapted to a particle sorting method based on particle drift as described above, comprising:
the calibration module is used for calibrating drift voltage of target particles in the classified particles;
the comparison and correction module is used for comparing the calibration drift voltage with a preset drift voltage, and correcting the drift voltage if the calibration drift voltage is smaller than the preset drift voltage until the calibration drift voltage is larger than the preset drift voltage;
the first micro-fluidic chip module is used for receiving the classified particles and performing electronic drift according to a preset drift voltage;
and the second microfluidic chip module is used for carrying out secondary sorting on the solution containing the drifting particles.
The device further comprises an upper computer, wherein the upper computer is in communication connection with the calibration module, the comparison and correction module, the first micro-fluidic chip module and the second micro-fluidic chip module, and is used for displaying the operation data of the calibration module, the comparison and correction module, the first micro-fluidic chip module and the second micro-fluidic chip.
Compared with the prior art, the invention has the beneficial effects that:
the method comprises the steps that a preset drift voltage is applied to a first micro-fluidic chip, so that particles needing to be separated and smaller than a target particle calibration drift voltage in classified particles are subjected to an external force F of the preset drift voltage, the particles deviate in the first micro-fluidic chip and impact in a passage in the first micro-fluidic chip, the particles needing to be separated are partially pretreated, and the secondary separation result is more accurate.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic block diagram of the present invention;
fig. 3 is a schematic view of a second microfluidic chip;
fig. 4 is a cross-sectional view of a second microfluidic chip.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Thus, the detailed description of the embodiments of the present invention provided in the following drawings is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention.
The traditional inertial microfluidic technology utilizes the fluid inertia effect to induce particles to move under the action of inertia force in a flow channel so as to realize accurate control, and has the advantages of simple flow channel structure, convenience in operation, high control precision and the like.
The particle drift means that if a particle with electric quantity q is acted by a constant uniform magnetic field B and other external forces F in a magnetic field, the particle moves in a direction perpendicular to the magnetic field B and the external forces F in addition to a spiral motion taking magnetic lines as axes, and the motion caused by the external forces is called drift, so that the particle can be shifted by adding an external force F outside the microfluidic chip, and the sorted particle can be pretreated.
Based on the above description, an embodiment of the present invention provides a particle sorting method based on particle drift as shown in fig. 1, including the following steps:
step 1: obtaining a calibration drift voltage of a target particle in the classified particles, comparing the calibration drift voltage with a preset drift voltage, if the calibration drift voltage is smaller than the preset drift voltage, executing the step 2, otherwise, executing the step 3;
step 2: correcting the drift voltage until the calibrated drift voltage is greater than the preset drift voltage;
and step 3: injecting the classified particles into the first microfluidic chip, and applying a drift voltage to the first microfluidic chip according to a preset drift voltage;
and 4, step 4: and collecting the drifting particles after drifting, and carrying out secondary sorting.
Applying a preset drift voltage to the first microfluidic chip to enable particles needing to be separated, which are smaller than the target particle calibration drift voltage, in the classified particles to be subjected to an external force F of the preset drift voltage, to deviate in the first microfluidic chip and impact in a passage in the first microfluidic chip, so that pretreatment on part of the particles needing to be separated is realized;
the invention is explained in detail below with reference to the drawings;
as shown in fig. 1, the first microfluidic chip is a microfluidic chip with a regular shape, such as a rectangle, a circle, a diamond, etc., in one embodiment, the first microfluidic chip is a rectangular hamburger structure, wherein the upper layer is an upper field voltage plate, the middle layer is a microfluidic chip, the lower layer is a lower field voltage plate, the upper field voltage plate and the lower field voltage plate are both connected with an external circuit to form a preset drift voltage, a drift channel is processed in the microfluidic chip, the cross section of the drift channel is a circle, and an uneven carbon nanotube coating is coated in the drift channel, on one hand, particles which are deflected can be captured, and on the other hand, the carbon nanotube coating does not affect the penetration of an external electric field, so the drift channel in this embodiment is coated with an uneven carbon nanotube coating, and the drift channel is a mosquito coil incense, on the one hand, the area of the first micro-fluidic chip can be reduced by the mosquito coil incense-shaped drift channel, on the other hand, the mosquito coil incense-shaped drift channel is provided with a plurality of arc-shaped curves, particles which are required to be separated and have the size far larger than the size of target particles can be enabled to be larger than the centrifugal force of a straight line due to the fact that the acceleration is too large at the arc-shaped curves, then the particles impact on the arc-shaped curves, pretreatment of the large-size particles is achieved, one end of the drift channel is a particle injection end, the other end of the drift channel is a collection end, the collection end is connected with a collection pool, wherein liquid used for decelerating and collecting floating particles is pre-filled in the collection pool, and supercooled water is selected as the liquid under the ordinary condition.
Referring to fig. 2, as shown in fig. 2, step 1: obtaining a calibration drift voltage of a target particle in the classified particles, comparing the calibration drift voltage with a preset drift voltage, if the calibration drift voltage is smaller than the preset drift voltage, executing a step 2, otherwise, executing a step 3, wherein the calibration drift voltage of the target particle is a voltage required by the target particle to generate maximum deviation under the action of an electric field, the maximum deviation is a distance corresponding to the target particle when the target particle impacts on a drift channel, the preset drift voltage is a maximum bearing voltage of a first micro-fluidic chip, injecting the target particle into the first micro-fluidic chip, loading the drift piezoelectric on the first micro-fluidic chip, detecting whether the target particle exists in a collection pool or not, when the target particle cannot be detected in the collection pool, the corresponding drift piezoelectric is the calibration drift voltage, and if the drift piezoelectric loaded on the first micro-fluidic chip reaches the maximum bearing voltage, when the target particles are still detected in the collection pool, the maximum bearing voltage is the calibration drift voltage;
step 2: and correcting the drift voltage until the calibrated drift voltage is larger than the preset drift voltage, specifically, S1: extracting a preset drift voltage value and a calibrated drift voltage value; s2: establishing a plane coordinate system and marking (a preset drift voltage value and a calibrated drift voltage value) as a first calculation point, and marking (a calibrated drift voltage value and a preset drift voltage value) as a second calculation point; s3: calculating the distance between the first calculation point and the second calculation point, and marking as a correction number; s4: subtracting the correction number from the preset drift voltage value to finish drift voltage correction, wherein a triangular graph of a first calculation point and a second calculation point is constructed, and the correction number can be quickly calculated through the Pythagorean theorem;
and step 3: and injecting the classified particles into the first micro-fluidic chip, applying a drift voltage to the first micro-fluidic chip according to a preset drift voltage, applying the preset drift voltage to the first micro-fluidic chip after the classified particles enter a drift channel in the first micro-fluidic chip through the injection end, and performing secondary separation on the drift particles in the collection pool after a preset time period.
Referring to fig. 3, as shown in fig. 3, the second microfluidic chip is composed of an upper substrate and a lower substrate; the upper layer substrate and the lower layer substrate are hermetically bonded together to form a second microfluidic chip; the upper substrate is provided with a liquid inlet hole, an inertia flow channel and a liquid outlet hole;
wherein, the liquid inlet hole and the liquid outlet hole are communicated with the outside and are used for leading in and leading out the solution containing the drift particles; the liquid inlet hole is communicated with the inertia flow channel and then divided into two branches, one branch is communicated with the first liquid outlet hole, and the other branch is communicated with the second liquid outlet hole.
The inertia runner is of an Archimedes spiral line structure, the inner diameter of the inlet of the runner is 10mm, and the outer diameter of the outlet of the runner is 30 mm. The cross section of the flow channel is rectangular, the width and the height are respectively 300 mu m and 50 mu m, and the width-height ratio of the flow channel is as follows.
In this embodiment, the upper substrate of the second microfluidic chip is fabricated using a standard soft lithography technique, and is made of polydimethylsiloxane, the lower substrate is a glass cover glass, and the upper substrate and the lower substrate are irreversibly bonded by an oxygen plasma cleaning process.
Referring to fig. 4, as shown in fig. 4, when performing the second sorting, first, the solution containing the drift particles is injected into the second microfluidic chip by using a precision syringe pump, and the flow rate is set to 450 μ L/min. The solution containing the drift particles enters the inertia flow channel through the liquid inlet hole and is in a random distribution state at the section A-A of the inlet of the inertia flow channel. Because the inertial flow channel is in an Archimedes spiral shape, microfluid in the flow channel generates two secondary flow vortexes which flow oppositely in the direction vertical to the main flow direction, and therefore the solution containing the drifting particles is simultaneously subjected to the inertial lift force generated by the wall surface induction of the inertial flow channel and the secondary flow drag force generated by the turning of the solution in the spiral flow channel in the flow channel. Then, the solution containing the drifting particles gradually generates an inertial focusing effect under the influence of an inertial lift force FL and a secondary flow drag force FD and transversely moves to different equilibrium positions, specifically, the equilibrium position of the large-size particles is slightly closer to the inner wall surface of the flow channel than the equilibrium position of the small-size particles, but the distance between the equilibrium positions of the two particles is smaller at the moment, and the two particles cannot be accurately sorted. Finally, the large-size particles are communicated and flow out through the first liquid outlet hole; small-size particle is through first going out the liquid hole intercommunication outflow, realizes that the secondary of unidimensional particle is selected separately, simultaneously because the first micro-fluidic chip of this application has carried out preliminary treatment to unidimensional particle not, so the application is when advancing the secondary and select separately, can be more accurate classify.
In addition to the above, the present application further provides a particle sorting system based on particle drift, for more accurately sorting particles of different sizes, comprising:
the calibration module is used for calibrating drift voltage of target particles in the classified particles;
the comparison and correction module is used for comparing the calibration drift voltage with a preset drift voltage, and correcting the drift voltage if the calibration drift voltage is smaller than the preset drift voltage until the calibration drift voltage is larger than the preset drift voltage;
the first micro-fluidic chip module is used for receiving the classified particles and performing electronic drift according to a preset drift voltage;
the second microfluidic chip module is used for carrying out secondary sorting on the solution containing the drifting particles;
the device comprises a calibration module, a comparison and correction module, a first micro-fluidic chip module and a second micro-fluidic chip module, and is characterized by further comprising an upper computer, wherein the upper computer is in communication connection with the calibration module, the comparison and correction module, the first micro-fluidic chip module and the second micro-fluidic chip module, and is used for displaying operation data of the calibration module, the comparison and correction module, the first micro-fluidic chip module and the second micro-fluidic chip.
In summary, the application provides a particle sorting method and system based on particle drift, by applying a preset drift voltage to a first micro-fluidic chip, particles needing to be separated, which are smaller than a target particle calibration drift voltage in the classified particles, are subjected to an external force F of the preset drift voltage, are deflected in the first micro-fluidic chip and impact on a passage in the first micro-fluidic chip, so that the particles needing to be separated are preprocessed, and the secondary sorting result is more accurate.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like, indicate orientations and positional relationships based on those shown in the drawings, and are used only for convenience of description and simplicity of description, and do not indicate or imply that the equipment or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be considered as limiting the present invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
The specific meanings of the above terms in the present invention can be understood in specific cases by those skilled in the art; the preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise forms disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (10)
1. A particle sorting method based on particle drift is characterized by comprising the following steps:
step 1: obtaining a calibration drift voltage of a target particle in the classified particles, comparing the calibration drift voltage with a preset drift voltage, if the calibration drift voltage is smaller than the preset drift voltage, executing the step 2, otherwise, executing the step 3;
step 2: correcting the drift voltage until the calibrated drift voltage is greater than the preset drift voltage;
and step 3: injecting the classified particles into the first microfluidic chip, and applying a drift voltage to the first microfluidic chip according to a preset drift voltage;
and 4, step 4: and collecting the drifting particles after drifting, and carrying out secondary sorting.
2. The method for sorting particles based on particle drift of claim 1, wherein the predetermined drift voltage is up to a maximum withstand voltage of the first microfluidic chip.
3. The method of claim 1, wherein the target particle has a nominal drift voltage that is the voltage required for the target particle to deflect maximally under the action of the electric field.
4. The particle sorting method based on particle drift according to claim 1, wherein the classified particles are injected into the first microfluidic chip, and a drift voltage is applied to the first microfluidic chip according to a preset drift voltage, specifically:
constructing a drift channel in the first microfluidic chip, wherein one end of the drift channel is a particle injection end, the other end of the drift channel is a collection end, and the collection end is connected with a collection pool;
and after the classified particles enter the drift channel in the first micro-fluidic chip through the injection end, applying a preset drift voltage to the first micro-fluidic chip, and after a preset time period, performing secondary separation on the drift particles in the collection pool.
5. The particle sorting method based on particle drift according to claim 1, wherein the calibration drift voltage of the target particles in the sorted particles is obtained, specifically, the target particles are injected into the first microfluidic chip, the drift piezoelectric is loaded on the first microfluidic chip, and whether the target particles exist in the collection pool is detected, until the target particles cannot be detected in the collection pool, the corresponding drift piezoelectric is the calibration drift voltage, and if the drift piezoelectric loaded on the first microfluidic chip reaches the maximum withstand voltage, and the target particles are still detected in the collection pool, the maximum withstand voltage is the calibration drift voltage.
6. The particle sorting method based on particle drift according to claim 1, wherein the performing drift voltage correction specifically comprises:
s1: extracting a preset drift voltage value and a calibrated drift voltage value;
s2: establishing a plane coordinate system and marking (a preset drift voltage value and a calibrated drift voltage value) as a first calculation point, and marking (a calibrated drift voltage value and a preset drift voltage value) as a second calculation point;
s3: calculating the distance between the first calculation point and the second calculation point, and marking as a correction number;
s4: and subtracting the correction number from the preset drift voltage value to finish the drift voltage correction.
7. The method of claim 1, wherein the secondary sorting comprises the steps of:
and injecting the solution containing the drifting particles in the collecting pool into the second microfluidic chip to complete secondary sorting.
8. The particle sorting method based on particle drift of claim 1, wherein the drift channel is mosquito coil shaped.
9. A particle sorting system based on particle drift, which is suitable for use in a particle sorting method based on particle drift according to any one of claims 1 to 8, and which comprises:
the calibration module is used for calibrating drift voltage of target particles in the classified particles;
the comparison and correction module is used for comparing the calibration drift voltage with a preset drift voltage, and correcting the drift voltage if the calibration drift voltage is smaller than the preset drift voltage until the calibration drift voltage is larger than the preset drift voltage;
the first micro-fluidic chip module is used for receiving the classified particles and performing electronic drift according to a preset drift voltage;
and the second microfluidic chip module is used for carrying out secondary sorting on the solution containing the drifting particles.
10. The particle sorting system based on particle drift of claim 9, further comprising an upper computer, wherein the upper computer is in communication connection with the calibration module, the comparison and correction module, the first microfluidic chip module and the second microfluidic chip module, and is used for displaying operation data of the calibration module, the comparison and correction module, the first microfluidic chip module and the second microfluidic chip.
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