CN214894705U - Multifunctional electrical detection chip for detecting cell parameters - Google Patents
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Abstract
The utility model discloses a multifunctional electrical detection chip for detecting cell parameters, which comprises runners connected in sequence through connecting runners; the two sides of the end part of the input flow channel, which is close to the connecting flow channel, are symmetrically provided with a primary conformal three-dimensional electrode pair; the two sides of the end part of the first-stage contraction flow channel, which are close to the connecting flow channel, are symmetrically provided with a second-stage conformal stereo electrode pair; one side of the first-stage contraction flow channel, which is close to the end part of the connecting flow channel, is provided with a three-stage conformal three-dimensional electrode; a four-stage conformal three-dimensional electrode is arranged on one side of the middle part of the secondary contraction flow channel; one side of the end part of the three-stage contraction flow channel, which is close to the connecting flow channel, is provided with a five-stage conformal three-dimensional electrode; six-stage conformal three-dimensional electrode pairs are symmetrically arranged on two sides of the end part of the three-stage contraction flow channel close to the connecting flow channel; the two sides of the end part of the output flow channel, which is close to the connecting flow channel, are symmetrically provided with seven-level conformal three-dimensional electrode pairs; the tail ends of all levels of conformal three-dimensional electrodes are conformal with the wall surface of the corresponding flow channel, and the tail ends of all levels of conformal three-dimensional electrodes are used as the wall surface of the corresponding flow channel.
Description
Technical Field
The utility model relates to a micro-fluidic field, especially a detect multi-functional electricity of cell parameter and detect chip.
Background
The living body single cell analysis is a core tool for revealing the behavior, the function and the microscopic heterogeneity of cells, can effectively clarify the basic structure, the functional information and the pathological state of the cells and reveal the individual difference among the cells, is a research hotspot in the fields of cell biology, developmental biology, disease pathology, cell quality control analysis, drug research and development, medical diagnosis and treatment and the like, and has important significance for revealing the rules of living bodies, exploring major disease mechanisms and the like. By constructing a micron-sized channel matched with the size of the cell, the microfluidic technology provides very convenient conditions for realizing efficient control and accurate detection of the cell.
The patent with the application number of 201910858617.X discloses a design method of a microfluidic chip, which can realize the capture of two single cells so as to realize the further analysis of cell characteristics. The patent application No. 202010226539.4 discloses a microfluidic chip capable of accurately acquiring information on intrinsic sizes of cells and nuclei through four T-shaped compression channels. Due to the limitations of the detection chip structure and the impedance measurement strategy, the existing microfluidic impedance cytometer can only analyze and process part of characteristic parameters, thereby affecting the reliability of cell detection. In addition, most of characteristic parameters extracted from the cell impedance peak can only indirectly represent the overall dielectric property and mechanical property of the cell, and cannot gain insight into the local information of the cell. Therefore, at present, an electrical detection chip capable of characterizing multiple parameters of the cells to be detected at one time is still lacking.
SUMMERY OF THE UTILITY MODEL
Utility model purpose: the utility model aims to solve the technical problem that to prior art not enough, provide a multi-functional electricity that can once only characterize a plurality of parameters of cell and detect chip.
In order to solve the technical problem, the utility model discloses the technical scheme who takes as follows:
a multifunctional electrical detection chip for detecting cell parameters comprises an input flow channel, a primary contraction flow channel, a primary recovery flow channel, a secondary contraction flow channel, a secondary recovery flow channel, a tertiary contraction flow channel and an output flow channel which are sequentially connected through a connecting flow channel;
the two sides of the end part of the input flow channel, which is close to the rear connecting flow channel, are symmetrically provided with a primary conformal three-dimensional electrode pair;
the two sides of the end part of the primary contraction flow channel, which is close to the front connection flow channel, are symmetrically provided with secondary conformal three-dimensional electrode pairs;
one side of the end part of the first-stage contraction flow channel, which is close to the rear connection flow channel, is provided with a three-stage conformal three-dimensional electrode;
one side of the middle part of the secondary contraction flow channel is provided with a four-stage conformal three-dimensional electrode;
one side of the end part of the three-stage contraction flow channel, which is close to the front connection flow channel, is provided with a five-stage conformal three-dimensional electrode;
the two sides of the end part of the three-stage contraction flow channel, which is close to the rear connection flow channel, are symmetrically provided with six-stage conformal three-dimensional electrode pairs;
the two sides of the end part of the output flow channel, which is close to the front connecting flow channel, are symmetrically provided with seven-level conformal three-dimensional electrode pairs;
the tail ends of all levels of conformal three-dimensional electrodes are conformal with the wall surface of the corresponding flow channel, and the tail ends of all levels of conformal three-dimensional electrodes are used as the wall surface of the corresponding flow channel.
Furthermore, the cross sections of the input runner, the first-stage contraction runner, the first-stage recovery runner, the second-stage contraction runner, the second-stage recovery runner, the third-stage contraction runner and the output runner, and connecting runners among the input runner, the first-stage contraction runner, the first-stage recovery runner, the second-stage contraction runner, the second-stage recovery runner, the third-stage contraction runner and the output runner are all circular;
the flow channel diameters of the input flow channel, the primary recovery flow channel, the secondary recovery flow channel and the output flow channel are the same and slightly larger than the diameter of the cell to be detected; the flow channel diameters of the first-stage contraction flow channel, the second-stage contraction flow channel and the third-stage contraction flow channel are the same and slightly smaller than the diameter of the cell to be detected.
Preferably, the three-level conformal stereo electrode and the five-level conformal stereo electrode are symmetrically distributed by taking the four-level conformal stereo electrode as a symmetry axis;
the three-level conformal stereo electrode and the five-level conformal stereo electrode are positioned on the same side, and the four-level conformal stereo electrode is positioned on the other side.
Specifically, the first-stage conformal stereo electrode pair and the seventh-stage conformal stereo electrode pair form a group of electrode pairs; the two-stage conformal stereo electrode pair and the six-stage conformal stereo electrode pair form a group of electrode pairs; the three-level conformal stereo electrode, the four-level conformal stereo electrode and the five-level conformal stereo electrode form a group of electrode pairs. After multi-frequency voltage signals are injected into the upper side electrodes of the chip in the three groups of electrode pairs, the lower side electrodes of the chip collect current signals and are processed by a difference circuit to obtain positive and negative wave signals, and after the positive and negative wave signals are filtered, signal segmented and impedance peak extraction and analysis, cell positions, cell diameters, cell shape parameters, impedance amplitudes, impedance phase parameters, cell impedance opacity (ratio of high-frequency impedance amplitudes to low-frequency impedance amplitudes) parameters and relaxation indexes of cells in different states can be obtained.
Preferably, the sealing material of each stage of flow channel is selected from any one of transparent materials such as PDMS, glass or PMMA; the material of each level of electrode is selected from any one of gold, ITO or nano silver and other conductive materials.
Has the advantages that:
the utility model discloses the circular cross section runner of multi-functional electricity detection chip is favorable to transporting, the extrusion deformation of cell to the symmetry of runner circular cross section is convenient for theoretical analysis and modeling simulation calculation more. The conformal three-dimensional electrode structure of the circumferential wall surface of the flow channel improves the volume fraction occupied by cells in the detection area, reduces the leakage of measurement electric signals, can effectively improve the sensitivity of cell impedance measurement, can perform one-time characterization of a plurality of parameters, and can play an important role in the development and clinical application of single cell analysis technology.
Drawings
These and/or other advantages of the present invention will become more apparent from the following detailed description of the present invention when taken in conjunction with the accompanying drawings and the following detailed description.
Fig. 1 is the utility model discloses multi-functional electricity detects chip overall structure sketch map.
Fig. 2 is a partial schematic view of the input flow channel and the end portion one-level conformal stereo electrode pair in the multifunctional electrical detection chip of the present invention.
FIG. 3 shows the signal detection result when the cell to be detected enters the first-level conformal stereo electrode pair.
FIG. 4 shows the signal detection result when the cell to be detected enters the position of the second-level conformal stereo electrode pair.
FIG. 5 shows the signal detection result when the cell to be detected enters the position of the three-level conformal stereo electrode.
Wherein each reference numeral represents:
11. an input flow channel, 12, a first contraction flow channel, 13, a first recovery flow channel, 14, a second contraction flow channel, 15, a second recovery flow channel, 16, a third contraction flow channel, 17, an output flow channel, 21, a first conformal three-dimensional electrode pair, 22, a second conformal three-dimensional electrode pair, 23, a third conformal three-dimensional electrode, 24, a fourth conformal three-dimensional electrode, 25, a fifth conformal three-dimensional electrode, 26, a sixth conformal three-dimensional electrode pair, 27, a seventh conformal three-dimensional electrode pair, 31, a first connecting flow channel, 32, a second connecting flow channel, 33, a third connecting flow channel, 34, a fourth connecting flow channel, 35, a fifth connecting flow channel, 36 and a sixth connecting flow channel.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings.
As shown in FIG. 1, the multifunctional electrical detection chip comprises an input flow channel 11, a primary contraction flow channel 12, a primary recovery flow channel 13, a secondary contraction flow channel 14, a secondary recovery flow channel 15, a tertiary contraction flow channel 16 and an output flow channel 17 which are connected in sequence through a connection flow channel.
Wherein, the input runner 11 is connected with the first-stage contraction runner 12 through a first connecting runner 31; the primary contraction flow passage 12 is connected with the primary recovery flow passage 13 through a second connecting flow passage 32; the first-stage recovery flow channel 13 is connected with the second-stage contraction flow channel 14 through a third connecting flow channel 33; the secondary contraction flow passage 14 is connected with the secondary recovery flow passage 15 through a fourth connecting flow passage 34; the second-stage recovery runner 15 is connected with the third-stage contraction runner 16 through a fifth connecting runner 35; the third-stage contracted flow passage 16 and the output flow passage 17 are connected by a sixth connecting flow passage 36.
The two sides of the end part of the input flow channel 11 close to the first connecting flow channel 31 are symmetrically provided with a first-stage conformal stereo electrode pair 21;
the two sides of the end part of the first-stage contraction flow channel 12 close to the first connection flow channel 31 are symmetrically provided with a second-stage conformal stereo electrode pair 22;
the first-stage contracted flow channel 12 is provided with a third-stage conformal three-dimensional electrode 23 at one side close to the end part of the second connecting flow channel 32;
a four-level conformal three-dimensional electrode 24 is arranged on one side of the middle part of the secondary contraction flow channel 14;
one side of the end part of the third-stage contracted flow channel 16 close to the fifth connecting flow channel 35 is provided with a fifth-stage conformal three-dimensional electrode 25;
the two sides of the end part of the third-stage contracted flow channel 16 close to the sixth connecting flow channel 36 are symmetrically provided with six-stage conformal three-dimensional electrode pairs 26;
the two sides of the end part of the output flow channel 17 close to the sixth connecting flow channel 36 are symmetrically provided with seven-stage conformal three-dimensional electrode pairs 27.
As shown in fig. 2, the ends of the first-order conformal stereo electrode pairs 21 are conformal with the corresponding wall surfaces of the input flow channel 11, and the ends of the first-order conformal stereo electrode pairs 21 serve as the wall surfaces of the input flow channel 11. The tail ends of other conformal three-dimensional electrodes are conformal with the wall surface of the corresponding flow channel, and the tail ends of the conformal three-dimensional electrodes are used as the wall surface of the corresponding flow channel.
The cross sections of the input runner 11, the first-stage contraction runner 12, the first-stage recovery runner 13, the second-stage contraction runner 14, the second-stage recovery runner 15, the third-stage contraction runner 16, the output runner 17 and connecting runners among the input runner, the first-stage contraction runner 12, the first-stage recovery runner 13, the second-stage contraction runner 14, the second-stage recovery runner 15, the third-stage contraction runner 16 and the output runner 17 are all circular; the flow channel diameters of the input flow channel 11, the primary recovery flow channel 13, the secondary recovery flow channel 15 and the output flow channel 17 are the same and are larger than the diameter of the cell to be detected; the flow channel diameters of the primary contracted flow channel 12, the secondary contracted flow channel 14 and the tertiary contracted flow channel 16 are the same and smaller than the diameter of the cell to be detected.
The three-level conformal stereo electrode 23 and the five-level conformal stereo electrode 25 are symmetrically distributed by taking the four-level conformal stereo electrode 24 as a symmetry axis; the three-level conformal stereo electrode 23 and the five-level conformal stereo electrode 25 are positioned on the same side, and the four-level conformal stereo electrode 24 is positioned on the other side.
The first-stage conformal stereo electrode pair 21 and the seven-stage conformal stereo electrode pair 27 form a group of electrode pairs to detect signals of cells in the whole detection area; the second-level conformal stereo electrode pair 22 and the sixth-level conformal stereo electrode pair 26 form a group of electrode pairs to detect signals of cells in the whole contraction flow channel; the three-level conformal stereo electrode 23, the four-level conformal stereo electrode 24 and the five-level conformal stereo electrode 25 form a group of electrode pairs to detect signals of the cell in the recovery deformation stage. After multi-frequency voltage signals are injected into the upper side electrodes of the chips in the three groups of electrode pairs, the lower side electrodes of the chips collect current signals and are processed by a difference circuit to obtain positive and negative wave signals, and after the positive and negative wave signals are filtered, signal segmented and impedance peak extraction and analysis, the cell position, cell diameter, cell shape parameters, impedance amplitude, impedance phase parameters, cell impedance opacity (ratio of high-frequency impedance amplitude to low-frequency impedance amplitude) parameters and relaxation index of the cells in different states can be obtained through one-time characterization.
The sealing material of each stage of flow channel is selected from PDMS transparent materials; the material of each stage of electrode is selected from gold conductive materials.
The input flow channel 11, the first-stage recovery flow channel 13, the second-stage recovery flow channel 15 and the output flow channel 17 of the multifunctional electrical detection chip in the embodiment have straight and round cross sectionsThe diameter is 30 micrometers, the diameters of cross-section circles of the first-stage contracted flow channel 12, the second-stage contracted flow channel 14 and the third-stage contracted flow channel 16 are 5 micrometers, the MCF-7 suspension is selected as the cell suspension, the MCF-7 suspension is injected into the multifunctional electrical detection chip through the input flow channel, after the MCF-7 suspension is filled in the cavity of the whole chip, the multifunctional electrical detection chip is connected with a pressure injector, a fixed value pressure is given through the pressure injector, the breast cancer cells are placed from the input flow channel, and the breast cancer cells move along with the MCF-7 suspension. The function signal generator is adjusted to generate an alternating excitation voltage with an amplitude of 1V and a frequency of 2 MHz. When the breast cancer cell moves in the input flow channel 11, an alternating current excitation voltage is applied to the first-level conformal stereo electrode pair 21 and the seventh-level conformal stereo electrode pair 27 to obtain a current response signal I1(Signal1) and I6(Signal6) as shown in FIG. 3. When the breast cancer cell moves to the head of the first-level contraction flow channel 13, excitation voltage is applied to the second-level conformal stereo electrode pair 22 and the sixth-level conformal stereo electrode pair 26 to obtain a current response signal I2(Signal2) and I5(Signal5) as shown in FIG. 4. When the breast cancer cell moves in the first-stage recovery flow channel 13, the second-stage contraction flow channel 14 and the second-stage recovery flow channel 15, an excitation voltage is applied to the four-stage conformal stereo electrode 24, and a current response signal I is obtained through the three-stage conformal stereo electrode 233(Signal3), obtaining a current response Signal I through the five-stage conformal stereo electrode 254(Signal4), as shown in FIG. 5, collect I1～I6The signals are processed by a differential circuit to obtain positive and negative wave signals, and then the signal conditioning stage is carried out. In the signal conditioning stage, baseline correction and denoising treatment are carried out on the original electrical impedance characteristic signal by using a moving average filter; in the signal segmentation stage, segmenting the conditioned signals by setting a threshold or reversely identifying idle time to obtain corresponding effective data when the cells pass through the sensing area; in the characteristic extraction stage, characteristic parameters such as peak value, peak width, passing time and the like are extracted from the segmentation signals by utilizing Gaussian fitting; and finally, in the characteristic processing stage, the influence brought by different positions of cells, fluid fluctuation and the like is considered, and the extracted characteristic parameters are further processed to improve the accuracy of measurement. Signal1 and Signal6The cell position, the cell diameter and the cell shape parameters of the breast cancer cells at the moment can be extracted after single and comprehensive analysis; after single and comprehensive analysis is carried out on Signal2 and Signal5, the time parameters of the last contraction and the first contraction of the breast cancer cells can be extracted, and the time ratio of the last contraction and the first contraction is the relaxation parameters of the quantitative cells; the impedance amplitude and the impedance phase parameters of the breast cancer cells can be extracted after single and comprehensive analysis is carried out on Signal3 and Signal 4; multiple frequency voltage signals can be injected to obtain cell impedance opacity parameters (ratio of high frequency impedance amplitude to low frequency impedance amplitude).
The utility model provides a thought and method of the multi-functional electricity of detecting cell parameter detects chip specifically realizes that this technical scheme's method and approach are many, above only the utility model discloses a preferred embodiment should point out, to ordinary technical personnel in this technical field, does not deviate from the utility model discloses under the prerequisite of principle, can also make a plurality of improvements and moist decorations, these improvements should also regard as with moist decorations the utility model discloses a protection scope. All the components not specified in the present embodiment can be realized by the prior art.
Claims (5)
1. A multifunctional electrical detection chip for detecting cell parameters is characterized by comprising an input flow channel (11), a first-stage contraction flow channel (12), a first-stage recovery flow channel (13), a second-stage contraction flow channel (14), a second-stage recovery flow channel (15), a third-stage contraction flow channel (16) and an output flow channel (17) which are sequentially connected through a connecting flow channel;
the two sides of the end part of the input flow channel (11) close to the rear connecting flow channel are symmetrically provided with a first-stage conformal stereo electrode pair (21);
the two sides of the end part of the primary contraction flow channel (12) close to the front connection flow channel are symmetrically provided with secondary conformal three-dimensional electrode pairs (22);
one side of the first-stage contraction flow channel (12), which is close to the end part of the rear connection flow channel, is provided with a three-stage conformal three-dimensional electrode (23);
one side of the middle part of the secondary contraction flow channel (14) is provided with a four-stage conformal three-dimensional electrode (24);
one side of the three-stage contraction flow channel (16) close to the end part of the front connection flow channel is provided with a five-stage conformal three-dimensional electrode (25);
the two sides of the end part of the three-stage contraction flow channel (16) close to the rear connecting flow channel are symmetrically provided with six-stage conformal three-dimensional electrode pairs (26);
the two sides of the end part of the output flow channel (17) close to the front connecting flow channel are symmetrically provided with seven-level conformal three-dimensional electrode pairs (27);
the tail ends of all levels of conformal three-dimensional electrodes are conformal with the wall surface of the corresponding flow channel, and the tail ends of all levels of conformal three-dimensional electrodes are used as the wall surface of the corresponding flow channel.
2. The multifunctional electrical detection chip for detecting cell parameters of claim 1, wherein the input flow channel (11), the primary contraction flow channel (12), the primary recovery flow channel (13), the secondary contraction flow channel (14), the secondary recovery flow channel (15), the tertiary contraction flow channel (16), the output flow channel (17), and the connection flow channels therebetween are all circular in cross section;
the flow channel diameters of the input flow channel (11), the primary recovery flow channel (13), the secondary recovery flow channel (15) and the output flow channel (17) are the same and are larger than the diameter of the cell to be detected; the flow channel diameters of the primary contraction flow channel (12), the secondary contraction flow channel (14) and the tertiary contraction flow channel (16) are the same and smaller than the diameter of the cell to be detected.
3. The multifunctional electrical detecting chip for detecting cell parameters as claimed in claim 1, wherein the three-level conformal stereo electrodes (23) and the five-level conformal stereo electrodes (25) are symmetrically distributed with the four-level conformal stereo electrodes (24) as a symmetry axis;
the three-level conformal stereo electrode (23) and the five-level conformal stereo electrode (25) are positioned on the same side, and the four-level conformal stereo electrode (24) is positioned on the other side.
4. The multifunctional electrical detecting chip for detecting cell parameters of claim 1, wherein the first-order conformal stereo electrode pair (21) and the seventh-order conformal stereo electrode pair (27) form a group of electrode pairs; the two-stage conformal stereo electrode pair (22) and the six-stage conformal stereo electrode pair (26) form a group of electrode pairs; the three-level conformal stereo electrode (23), the four-level conformal stereo electrode (24) and the five-level conformal stereo electrode (25) form a group of electrode pairs.
5. The multifunctional electrical detection chip for detecting cell parameters according to claim 1, wherein the sealing material of each stage of flow channel is selected from any one of PDMS, glass or PMMA; the material of each stage of electrode is selected from any one of gold, ITO or nano silver.
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