CN110938534B - Passive wireless cell sorting system - Google Patents
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Abstract
The invention discloses a passive wireless cell sorting system, and belongs to the technical field of microfluidic chips. The system comprises an external signal transmitting part and a passive wireless cell sorting chip part; the passive wireless cell sorting chip part comprises a substrate and a microfluidic channel part; the substrate part comprises an LC resonance circuit comprising a sensitive capacitor, a rectifying circuit and a cantilever beam switch; the upstream part of the microfluidic channel is positioned between two poles of the sensitive capacitor, and the cantilever beam switch is positioned at the upstream and downstream branches of the microfluidic channel. The system utilizes the passive wireless sorting chip, so that the sorting process can be kept in a constant-temperature incubator without influencing the growth of cells; meanwhile, the sorting chip applies a capacitance sensitive sorting principle, so that the sorting accuracy and universality can be effectively guaranteed.
Description
Technical Field
The invention discloses a passive wireless cell sorting system, and belongs to the technical field of microfluidic chips.
Background
The microfluidic technology can accurately control fluid and organisms under submicron scale, and can be interactively applied to multiple subjects such as physics, chemistry, biochemistry, biology and the like to form a laboratory on a chip. The micro-structure in the micro-fluidic chip enables the micro-fluidic chip to have wide application, and the micro-scale structures such as micro-columns, micro-holes, micro-valves, micro-channels, micro-electrodes and the like enable the micro-fluidic chip to have great flexibility and operability in design and use. Different control effects can be achieved by different microstructure designs.
Cell sorting technology refers to a technique in which a cell is separated from a multicellular sample. Cell sorting is a biological experimental method frequently used in subjects such as diagnostic tests, pathological studies and the like. The traditional biochemical sorting method is to fix cells inside a chip by combining specific antigens on the surfaces of the cells with antibodies fixed on the inner walls of flow channels so as to achieve the purpose of sorting the cells, but the method can destroy the physiological state of the cells and make the captured cells difficult to release. The electrical method includes an electrode impedance detection method, a dielectrophoresis method, and the like. The electrode impedance detection method utilizes the impedance between electrodes as the basis of sorting, namely the impedance difference generated when cells pass through the electrodes is used as a sorting mark, the method requires that the electrodes enter cell suspension to be detected, and the electrodes are used as a wired connection detection mode, and most samples need to be taken out from an original constant temperature incubator. The dielectrophoresis cell sorting technology is a technology for inducing different dipole moments by different types of cells by utilizing a non-uniform electric field area manufactured by using a microelectrode so as to realize cell track control, but the method has higher requirements on the structure and the manufacturing process of the microelectrode, and particularly has greater challenge on generating effective electric field gradient in the whole channel depth.
Disclosure of Invention
The invention aims to provide a passive wireless cell sorting system aiming at the defects of the background technology, which provides energy for an execution switch on a sorting chip in a wireless energy coupling mode, realizes high-throughput cell screening in a passive wireless mode, and solves the technical problems that the biochemical sorting technology destroys the physiological state of cells, the wired electrical detection technology interrupts the culture environment, and the dielectrophoresis sorting technology has higher requirements on the structure and the manufacturing process of a microelectrode.
The invention adopts the following technical scheme for realizing the aim of the invention:
a passive wireless cell sorting system includes an external signal transmitting section and a passive wireless cell sorting chip section; the external signal transmitting part comprises a signal generator and a first inductor, and the first inductor is connected with the signal generator in series; the passive wireless cell sorting chip part comprises a substrate part and a micro-fluid channel part from bottom to top; the substrate part comprises a second inductor, a sensitive capacitor, a rectifying circuit and a cantilever beam switch; the micro-fluid channel part comprises a cell suspension inlet, a cell micro-channel positioned at the upstream, a target cell micro-channel positioned at the branch downstream, a target cell outlet, a residual cell channel and a residual cell outlet.
The second inductor is connected with the sensitive capacitor in series to form an LC resonance loop, and the rectifying circuit is connected with the cantilever beam switch in series and then connected with the sensitive capacitor in parallel. The first inductor is axially aligned with the second inductor. The second inductor is a plane inductor; the sensitive capacitor is a parallel plate plane capacitor; the cantilever beam switch comprises a movable end and a fixed end; the cell micro-channel is positioned between two electrodes of the sensitive capacitor; the movable end of the cantilever beam switch is positioned in the micro-fluid channel and is positioned at the inlet of the target cell micro-fluid channel at the downstream of the branch; the fixed end of the cantilever switch is located outside the microfluidic channel and is aligned parallel to the movable end of the cantilever switch.
The sensing system comprises the following steps:
the first step is as follows: the signal generator transmits electromagnetic waves with specific frequency through the first inductor; the second inductor couples energy in a mutual inductance coupling mode, and applies voltage generated by coupling to the cantilever beam switch after passing through the rectifying circuit;
the second step is that: a cell sample to be selected enters from a cell suspension inlet, flows into a cell micro-channel and passes between two polar plates of a sensitive capacitor, and the specific size, dielectric constant and intracellular capacitance of a target cell can cause the capacitance value of the sensitive capacitor to change, so that the resonance frequency of an LC resonance circuit is changed;
The third step: when the target cell passes through the sensitive capacitor, the capacitance change causes the resonance frequency of the LC series resonance circuit to be consistent with the frequency of the signal transmitted by the first inductor, and the energy which can be coupled by the second inductor reaches the maximum, so that the voltage applied to the two ends of the cantilever beam switch reaches the threshold voltage of the switch opening; the movable end of the cantilever beam switch is driven by the action of electrostatic force to move towards the fixed end until the movable end touches the edge of the fluid channel to stop; at the moment, the channel port of the target cell micro-channel is opened, and the inlet and outlet of the rest cell channels are closed due to the prevention of the movable end of the cantilever beam; the target cell successfully enters a channel of the target cell micro-channel and finally leaves from a target cell outlet; the movable end of the cantilever beam restores to the initial state after relaxation time, at the moment, the channel port of the target cell micro-channel is closed, the inlet and outlet of the rest cell channels are in an open state, new cells to be detected enter from the cell suspension inlet, and a new round of monitoring is started;
the fourth step: when the non-target cell passes through the sensitive capacitor, the sensitive capacitance value is not in a specific range, and the energy which can be coupled by the second inductor cannot reach the threshold condition of opening the cantilever beam switch. Therefore, the movable end of the cantilever beam keeps still, at the moment, the channel port of the target cell micro-channel is closed due to the existence of the movable end, and the inlet and outlet of the residual cell channel are kept smooth, so that non-target cells smoothly enter the residual cell channel and finally leave the chip through the residual cell outlet.
It is worth noting that the time for the cell sample to flow from the sensitive capacitor to the branch of the microfluidic channel is the same as the relaxation time of the cantilever beam switch, so that the target cell can have enough time to enter the target cell microfluidic channel after leaving the sensitive capacitor. And the width of the micro flow channel is smaller than the length of the movable end of the cantilever beam switch so as to ensure that the movable end can completely close the rest cell flow channel.
By adopting the technical scheme, the invention has the following beneficial effects:
(1) according to the passive wireless cell sorting system provided by the invention, energy is obtained through mutual inductance coupling of the external signal transmitting part and the LC resonant circuit, and the cantilever beam switch is controlled to be opened and closed after rectification, so that the flow direction of target cells is controlled, and the sorting function of the target cells is finally realized.
(2) The capacitance-sensitive sorting principle is utilized, and the method has the advantages of accuracy and universality: different types of target cells have corresponding characteristics of size, dielectric constant, intracellular capacitance and the like, and the characteristics can cause the change of the sensitive capacitance value, so that the sorted target types are diversified, and the result is more accurate.
Drawings
FIG. 1 is a diagram of a system of the present invention.
FIG. 2 is a detailed view of the cantilever switch in the presence of a target cell.
FIG. 3 is a detailed view of the cantilever switch in the presence of non-target cells.
The reference numbers in the figures illustrate: 1. an external signal transmitting part, 2, a passive wireless cell sorting chip part, 11, a signal generator, 12, a first inductor, 21, a substrate, 22, a microfluidic channel part, 211, a second inductor, 212, a sensitive capacitor, 213, a rectifying circuit, 214, a cantilever beam switch, 2141, a movable end, 2142, a fixed end, 221, a cell suspension inlet, 222, a cell microchannel, 223, a target cell microchannel, 224, a target cell outlet, 225, a residual cell microchannel, 226, and a residual cell outlet.
Detailed Description
The technical scheme of the invention is explained in detail in the following with reference to the attached drawings.
As shown in fig. 1, a passive wireless cell sorting system includes an external signal transmitting section 1 and a passive wireless cell sorting chip section 2. The external signal transmitting part 1 is arranged outside the constant-temperature incubator, the passive wireless cell sorting chip part 2 is arranged in the incubator, and the whole sorting link cannot interrupt the cell culture environment.
The external signal transmitting section 1 includes: signal generator 11 and first inductance 12, first inductance 12 is connected signal generator 11 in series. The passive wireless cell sorting chip part 2 comprises a substrate 21 and a microfluidic channel part 22 from bottom to top, wherein the substrate part is a glass substrate, the microfluidic channel is prepared from a PDMS material, and the microfluidic channel made of the PDMS material is bound with the substrate part of the glass substrate after being treated by oxygen and other particles. The substrate 21 is provided with a second inductor 211, a sensitive capacitor 212, a rectifying circuit 213 and a cantilever switch 214. The second inductor 211 is a planar inductor, the sensitive capacitor 212 is a parallel-plate planar capacitor, and the cantilever switch 214 includes a movable end 2141 and a fixed end 2142. The second inductor 211 and the sensitive capacitor 212 are connected in series to form an LC resonant circuit, a series branch consisting of a rectifying circuit 213 and a cantilever beam switch 214 is connected in parallel between two poles of the sensitive capacitor 212, and the first inductor 12 is axially aligned with the second inductor 211.
The microfluidic channel part 22 is prepared with a microfluidic channel, and liquid PDMS is poured into a 3D mold with a microfluidic channel pattern and then cured at high temperature to form the microfluidic channel with a cell suspension inlet 221, a cell micro-channel 222 located at the upstream, a target cell micro-channel 223 located at the branch downstream, a target cell outlet 224, a residual cell micro-channel 225 and a residual cell outlet 226. The upstream cell microchannel 222 is located between the two electrodes of the sensing capacitor 212. As shown in fig. 2 and 3, the movable end 2141 of the cantilever switch is located in the surplus-cell microchannel 225 and at the inlet of the target-cell microchannel 223 downstream of the branch; fixed end 2142 of the cantilever switch is located outside the microfluidic channel and is aligned parallel to the movable end 2141 of the cantilever switch in the closed state.
Sorting cells using the system of the present application includes the following four steps.
The first step is as follows: the signal generator 11 transmits electromagnetic waves with specific frequency through the first inductor 12, and the signal transmitted by the signal transmitter is a frequency sweeping signal containing the working frequency of the LC resonance circuit when the target cell sample to be detected flows through the upstream cell micro-channel 222; the second inductor 211 couples energy by mutual inductance coupling, and the rectifying circuit 213 applies the voltage generated by coupling to the cantilever switch 214.
The second step is that: the cell sample to be selected enters from the cell suspension inlet 221, flows into the cell micro-channel 222, passes through the space between the two plates of the sensitive capacitor 212, and the capacitance value of the sensitive capacitor 212 is changed due to the specific size, dielectric constant and intracellular capacitance of the target cell, so that the resonance frequency of the LC resonance circuit is changed.
The third step: when the capacitance change caused by the target cell a passing through the sensitive capacitor 212 makes the resonant frequency of the LC series resonant tank consistent with the frequency of the signal transmitted by the first inductor 12, at this time, the energy that can be coupled by the second inductor 211 reaches the maximum, so that the voltage applied to the two ends of the cantilever switch 214 reaches the threshold voltage of the switch opening; the movable end 2141 of the cantilever switch 214 is driven by electrostatic force to move toward the fixed end 2142 until it touches the edge of the remaining cell microchannel 225, as shown in FIG. 2; at this time, the channel port of the target cell microchannel 223 is opened, and the inlet and outlet of the remaining cell microchannel 225 are closed by the blocking of the cantilever movable end 2141; the target cell a successfully enters the channel of the target cell micro flow channel 223 and finally exits from the target cell outlet 224, as indicated by the dotted arrow in fig. 2; the cantilever movable end 2141 returns to its initial state after the relaxation time, at this time, the channel port of the target cell microchannel 223 is closed, the inlet and outlet of the remaining cell microchannel 225 are opened, and a new cell to be detected enters from the cell suspension inlet 221, and a new round of monitoring is started. The time for the cell sample to be detected to flow from the microfluidic channel between the two poles of the sensitive capacitor 212 to the branch of the microfluidic channel is the same as the relaxation time of the cantilever beam switch 214, so that the target cell can have enough time to enter the target cell microfluidic channel 223 after leaving the sensitive capacitor 212.
The fourth step: when the non-target cell B passes through the sensing capacitor 212, the sensing capacitance is not within a specific range, and the energy that can be coupled by the second inductor 211 fails to reach the threshold condition for turning on the cantilever switch 214. Thus, the cantilever movable end 2141 remains stationary, as shown in FIG. 3. At this time, the channel opening of the target cell microchannel 223 is closed due to the existence of the movable end 2141, and the inlet and outlet of the remaining cell microchannel 225 are kept open, so that the non-target cells smoothly enter the remaining cell microchannel 225 and finally leave the chip through the remaining cell outlet 226, as shown by the dotted arrow in fig. 3. The width of the microfluidic channel should be smaller than the length of the movable end 2141 of the cantilever switch to ensure that the movable end 2141 can completely close the remaining cell microfluidic channel 225.
Claims (6)
1. A passive wireless cell sorting system is characterized by comprising an external signal transmitting part and a passive wireless cell sorting chip part, wherein the passive wireless cell sorting chip part comprises a substrate part and a microfluidic channel part bonded on the substrate part, the substrate part is provided with an LC resonance circuit comprising a sensitive capacitor, a rectifying circuit and a cantilever beam switch, a series branch consisting of the rectifying circuit and the cantilever beam switch is connected between two poles of the sensitive capacitor in parallel, the microfluidic channel part forms a microfluidic channel which is provided with a cell suspension inlet, a cell microchannel positioned at the upstream, a target cell microchannel positioned at the downstream of a bifurcation, a target cell outlet, a residual cell microchannel and a residual cell outlet, the cell microchannel positioned at the upstream is positioned between two electrodes of the sensitive capacitor, the movable end of the cantilever beam switch is positioned in the residual cell microchannel and is positioned at the inlet of the target cell microchannel at the downstream of the bifurcation, the fixed end of the cantilever beam switch is positioned outside the micro-fluid channel and is aligned with the movable end of the cantilever beam switch in parallel in a closed state; the external signal transmitting part comprises a signal generator and a first inductor connected in series with the signal generator, the signal transmitted by the signal generator is a frequency sweeping signal containing the working frequency of the LC resonance circuit when the cell sample to be detected flows through the cell micro-channel positioned at the upstream, and the first inductor is axially aligned with the inductor in the LC resonance circuit.
2. The system of claim 1, wherein the relaxation time of the cantilever switch is the same as the time for the cell sample to flow from the upstream microfluidic channel to the branch of the microfluidic channel.
3. The system of claim 1, wherein the width of the microfluidic channel is less than the length of the movable end of the cantilever switch.
4. The passive wireless cell sorting system according to claim 1, wherein the substrate portion is a glass substrate, the microfluidic channel is made of a PDMS material, and the microfluidic channel made of the PDMS material is bonded to the substrate portion of the glass substrate after the oxygen plasma treatment.
5. The system of claim 1, wherein the microfluidic channel is formed by pouring liquid PDMS into a 3D mold with a microfluidic channel pattern and curing at high temperature.
6. A passive wireless cell sorting method is realized based on the system of any one of claims 1 to 5, and is characterized in that the working frequency range of an LC resonance circuit is determined according to the sensitive capacitance change condition caused by a cell sample to be detected, the frequency of a transmitting signal generated by an external signal transmitting part is adjusted to enable the frequency band of the transmitting signal to comprise the working frequency range of the LC resonance circuit, the cell sample to be detected is added into a cell suspension inlet, the outflow conditions of cell suspensions at a target cell outlet and a residual cell outlet are observed, the cell sample contained in the cell suspension flowing out of the target cell outlet is the target cell, and the cell sample contained in the cell suspension flowing out of the residual cell outlet is the non-target cell.
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