CN109164149B - Electrochemical biosensor for online detection of hydrogen peroxide in cells based on combination of paper chip and screen-printed electrode - Google Patents
Electrochemical biosensor for online detection of hydrogen peroxide in cells based on combination of paper chip and screen-printed electrode Download PDFInfo
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Abstract
The invention discloses an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen printing electrode. The prepared nano platinum/multi-wall carbon nanotube material is modified on the surface of a screen printing electrode and is organically combined with a self-made integrated cell culture platform-paper-based chip to manufacture the biosensing device for monitoring the release of hydrogen peroxide from PC12 cells (adrenal pheochromocytoma cells) on line. The device is connected with an electrochemical workstation, realizes the online monitoring of the activities of PC12 cells and Alzheimer's Disease (AD) model cells, has good specificity and high sensitivity, and provides a new method for the research of related fields such as cell activity monitoring.
Description
Technical Field
The invention relates to an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen printing electrode, which belongs to the technical field of detection, and the paper chip can be used for detection of hydrogen peroxide in PC12 cells and AD model cells.
Background
The most common age-related of neurodegenerative diseases is alzheimer's disease, the pathogenic mechanism of which involves the aggregation and deposition of beta amyloid. The research shows that the accumulation and overload of beta amyloid (A beta) can cause oxidative stress reaction, promote the inner flow of calcium ions in cells, and lead to the depolarization of mitochondrial membranes so as to increase the expression level of hydrogen peroxide in the cells. Therefore, the detection of the content of the hydrogen peroxide molecular compound at the cellular level is of great significance for understanding the pathological mechanism research of the age-related neurodegenerative diseases.
In recent years, electrochemical methods have become a hot point of research for scientists due to their unique advantages, such as low economic cost, high sensitivity, good selectivity, etc. However, the conventional electrode has the disadvantages of large sample amount, complex operation, difficulty in integration and miniaturization, and the like, and is not favorable for realizing online monitoring of the cell level. The advent of screen printed electrodes made this problem readily apparent. The screen printing electrode is simple to manufacture, can be produced in batches, is low in price, convenient to carry and small in required sample amount, and is very suitable for being combined with a paper chip.
The invention discloses an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip and a screen-printed electrode, wherein the screen-printed electrode is modified by using a prepared nano platinum/multi-wall carbon nanotube material, and a novel method for online detection of hydrogen peroxide released by PC12 cells is established by combining a self-made integrated cell culture platform-paper-based chip, and is used for online monitoring of activities of PC12 cells and AD model cells.
Disclosure of Invention
1. The invention aims to establish an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a silk-screen printing electrode.
The invention aims to realize the electrochemical biosensor for detecting the hydrogen peroxide in the cells on line based on the combination of the paper chip and the silk-screen printed electrode.
The paper chip is of a three-layer structure, the upper layer is a polydimethylsiloxane elastomer layer, and a round hole is formed in the middle of the polydimethylsiloxane elastomer layer and is used as a cell culture pool and a detection sample groove; the middle layer is a filter paper layer which is used as a carrier for culturing cells; the lower layer is a polydimethylsiloxane elastomer layer without a round hole and used as a carrier of the paper chip, and the three layers are tightly fixed by a clamp; the filter paper layer adopts the filter paper processed by the following method: heating and boiling filter paper to remove water-soluble impurities, soaking the filter paper in an EDTA-Na solution to complex metal ions in the paper base, soaking the filter paper in ethanol to remove organic impurities, modifying the filter paper with chitosan, and airing the filter paper for later use.
The hydrogen peroxide concentration is in a good linear relation with the current response value in the range of 0.1-8 mu mol/L, and the linear equation I (mu A) = 0.0538 + 0.1585CH2O2 (μmol/L)，R2 = 0.9944, detection limit 0.0232 μmol/L.
The invention discloses a method for detecting hydrogen peroxide in cells based on an electrochemical biosensor for detecting hydrogen peroxide in cells on line by combining a paper chip and a screen printing electrode, which is characterized by comprising the following steps of:
(1) designing and manufacturing a paper chip: the paper chip is of a three-layer structure, the upper layer is a polydimethylsiloxane elastomer layer, and a round hole is formed in the middle of the polydimethylsiloxane elastomer layer and is used as a cell culture pool and a detection sample groove; the middle layer is a filter paper layer which is used as a carrier for culturing cells; the lower layer is a polydimethylsiloxane elastomer layer without a round hole and used as a carrier of the paper chip, and the three layers are tightly fixed by a clamp; the filter paper layer adopts the filter paper processed by the following method: heating and boiling filter paper to remove water-soluble impurities, then soaking the filter paper in an EDTA-Na solution to complex metal ions in the paper base, then soaking the filter paper in ethanol to remove organic impurities, finally modifying the filter paper with chitosan, and airing the filter paper for later use; (2) determination of hydrogen peroxide in cells: taking the cell suspension, placing the cell suspension in the paper chip prepared in the step (1), placing the paper chip in a culture dish, and placing the culture dish in 5wt% CO2In an incubator at 37 ℃ andchanging the culture solution in a place close to water every two days; after the culture is finished, a phorbol ester solution is used for stimulating cells to generate hydrogen peroxide, the paper chip and a screen printing electrode of the modified nano platinum/multi-wall carbon nanotube material are used as working electrodes to pass through an electrochemical workstation together, and the hydrogen peroxide in the cells is detected by adopting a time current method.
When the growth state of the PC12 cells reaches 80% of that of the culture bottle and is observed to be good, the culture solution is poured out and washed by phosphate buffer solution, contacting cells with a proper amount of PBS solution, placing the cells in an incubator for culture, collecting PBS buffer solution contacting PC12 cells, preparing hydrogen peroxide solutions with a series of concentrations by using the solutions, placing the hydrogen peroxide solutions on a paper chip, recording current response values of hydrogen peroxide with different concentrations under the conditions that the voltage is-0.3V and the time is 50 s by using a time-current method together with a screen printing electrode of a modified nano platinum/multi-wall carbon nanotube material, in the cell-contacted PBS buffer, the current response signal increased with the increase of the hydrogen peroxide concentration, which is in a good linear relationship with the current response value in the range of 0.1-8 μmol/L, and the linear equation I (μ a) = 0.0538 + 0.1585C.H2O2 (μmol/L)，R2 = 0.9944, detection limit 0.0232 μmol/L (S/N = 3).
The invention discloses a method for detecting PC12 cells and AD model cells by using an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip and a silk-screen printing electrode, which is characterized in that the paper chip is used as a PC12 cell culture and detection carrier, and a nano platinum/multi-walled carbon nanotube modified printing electrode is used as a working electrode to realize online activity detection of the PC12 cells and the AD model cells.
Taking PC12 cells in logarithmic growth phase, digesting, centrifuging, re-suspending with complete culture solution, and counting to 5 × 104And each/mL is planted in a paper-based chip, and the online activity detection of PC12 cells and AD model cells is realized by using a time-current method by using a nano platinum/multi-wall carbon nanotube modified printing electrode as a working electrode.
2. The paper chip provided by the invention uses filter paper as a substrate to perform cell culture, and detects the level of hydrogen peroxide in cells by establishing an electrochemical sensor of a nano platinum/multi-walled carbon nanotube modified printed electrode.
3. The invention discloses an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen printing electrode, which sequentially comprises the following steps:
(1) preparation of the electrodes
Cleaning a Screen Printing Carbon Electrode (SPCE) for 3 times by using deionized water, drying by using nitrogen, dripping multi-walled carbon nanotube (MWCNTs) material dispersion liquid on the surface of a working electrode of the screen printing carbon electrode, naturally drying, dripping 2 mmol/L chloroplatinic acid solution and 0.5 mol/L sulfuric acid solution, and depositing platinum nanoparticles (PtNPs) on the surface of the working electrode of the screen printing carbon electrode by adopting a cyclic voltammetry method. Finally, nafion is dripped in the solution and naturally dried to form a nafion membrane, thus obtaining nafion/PtNPs/MWCNTs/SPCE.
(2) Design and manufacture of paper chip
The paper chip is of a three-layer structure, the upper layer structure is 2 cm multiplied by 0.3 cm of polydimethylsiloxane elastomer (PDMS), and a round hole with the diameter of 1 cm is arranged in the middle of the PDMS to be used as a cell culture pool and a detection sample groove; the middle structure is filter paper with the diameter of 8 mm and is used as a carrier for culturing cells; the lower layer structure is a non-porous PDMS structure with the thickness of 2 cm multiplied by 0.3 cm and is used as a carrier of the paper chip, and the three layers of structures are tightly fixed by a clamp.
(3) Treatment of filter paper
Heating and boiling filter paper to remove water-soluble impurities, soaking the filter paper in a 1 mol/L EDTA-Na solution to complex metal ions in the paper base, soaking the filter paper in ethanol to remove organic impurities, modifying the filter paper with 0.5% chitosan, and airing the modified filter paper for later use.
(4) Culture of cells on paper chip and detection of hydrogen peroxide
Placing the cell suspension in a self-made paper chip, placing the whole device in a culture dish, and placing the culture dish in 5% CO2The culture medium was changed at a place near water in the 37 ℃ incubator every two days. Fixing the paper chip and modified screen-printed electrode in a self-made device (shown in figure 1), and stimulating cells with phorbol ester solutionAnd generating hydrogen peroxide, setting parameters of an electrochemical workstation, and detecting the hydrogen peroxide in the cells by adopting a time current method.
The invention has the advantages that:
the nano platinum/multi-wall carbon nanotube material modified printed electrode has the advantages of high specificity and strong sensitivity when being used for detecting hydrogen peroxide. The paper chip has the unique advantages of low cost, environmental protection, miniaturization and portability, and can be used for culturing paper-based cells and used as a matrix for detecting hydrogen peroxide released by the cells.
Drawings
FIG. 1 is a working principle diagram of an electrochemical biosensor for detecting hydrogen peroxide released by PC12 cells based on online detection of hydrogen peroxide in cells by a paper chip combined with a silk-screen printed electrode. (in the figure: 1: hydrogen peroxide detection box; 2: cell culture tank; 3: printed electrode linking port; 4: printed electrode; 5: electrochemical workstation; 6: reference electrode; 7: working electrode; 8: counter electrode; 9: paper-based carrier; 10: closed PDMS layer; 11: perforated PDMS layer; 12: PC12 cell).
FIG. 2 is a scanning electron microscope representation of a bare electrode and a gradually modified nanomaterial electrode in an electrochemical biosensor for on-line detection of hydrogen peroxide in cells based on a paper chip combined with a silk screen printed electrode according to the present invention.
FIG. 3 is a linear relationship diagram of the electrochemical biosensor for online monitoring of PC12 cells based on online detection of hydrogen peroxide in cells by combining a paper chip and a silk-screen printed electrode.
FIG. 4 is a graph showing the result of online monitoring of AD model cells by an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen printed electrode according to the present invention.
Detailed Description
In order to make the technical problems, technical solutions and effects to be solved by the present invention clearer, the present invention is described in further detail below with reference to embodiments and drawings.
As shown in FIG. 1, the working principle diagram of the electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a silk-screen printed electrode according to the present invention is as follows: the nano platinum/multi-wall carbon nanotube modified screen printing electrode (nafion/PtNPs/MWCNTs/SPCE) is inserted into a paper chip/PDMS hybrid device, then an electrochemical workstation and a computer are connected, and the modified screen printing electrode (as shown in figure 1, 6: a reference electrode; 7: a nano platinum/multi-wall carbon nanotube working electrode; 8: a counter electrode) is used for detecting hydrogen peroxide released by cells by adopting a time current method.
Example 1:
(1) preparation of nafion/PtNPs/MWCNTs/SPCE electrode
And cleaning the screen printing carbon electrode for 3 times by using deionized water, drying by blowing nitrogen, dripping the dispersion liquid of the multi-wall carbon nanotube material on the surface of the working electrode of the screen printing carbon electrode, naturally drying, dripping 2 mmol/L chloroplatinic acid solution and 0.5 mol/L sulfuric acid solution, and depositing the platinum nanoparticles on the surface of the working electrode of the screen printing carbon electrode by adopting a cyclic voltammetry method. Finally, nafion is dripped in the solution and naturally dried to form a nafion film, thus obtaining the nano platinum/multi-wall carbon nanotube modified screen printing electrode (nafion/PtNPs/MWCNTs/SPCE) (reference number 7 in figure 1: as a working electrode). As shown in fig. 1, a reference electrode 6, a working electrode 7 and a counter electrode 8 are integrated in the screen-printed carbon electrode 4, the reference electrode 6 is integrated on one side of the working electrode 7 on the surface of the screen-printed carbon electrode 4, and the counter electrode 8 is integrated on the other side of the working electrode 7.
As shown in fig. 2, A, B and C in the figure are respectively a scanning electron microscope characterization figure of a bare screen printing carbon electrode, a multi-walled carbon nanotube modified screen printing carbon electrode (MWCNTs/SPCE) and a platinum nano/multi-walled carbon nanotube modified screen printing carbon electrode (PtNPs/MWCNTs/SPCE), and it can be known from a in fig. 2 that the surface of the screen printing electrode is composed of granular carbon powder and is coarse, and when the surface of the screen printing electrode is modified by MWCNTs (B in fig. 2), the MWCNTs with tubular structures are uniformly dispersed on the surface of the modified electrode; in fig. 2, C shows that after the platinum nanoparticles are further modified to the surface of the electrode, flocculent platinum nanoparticles appear on the surface of the multi-walled carbon nanotube, and the average size of the platinum nanoparticles is about 40 nm.
(2) Design and manufacture of paper chip
The paper chip is of a three-layer structure, the upper layer structure is a polydimethylsiloxane elastomer layer (PDMS) with the diameter of 2 cm multiplied by 0.3 cm, and a round hole with the diameter of 1 cm is arranged in the middle of the PDMS layer (as shown in figure 1, 11: the PDMS layer with holes) and is used as a cell culture pool and a detection sample groove; the middle layer structure is a filter paper layer (9: paper-based carrier) with the diameter of 8 mm, and the filter paper layer is used as a carrier for culturing cells; the lower layer structure is a non-porous PDMS layer structure (11: a sealed PDMS layer) with the thickness of 2 cm multiplied by 0.3 cm, and the three-layer structure is used as a carrier of the paper chip and is tightly fixed by a clamp to form a paper chip/PDMS hybrid device.
(3) Treatment of filter paper
And (3) treating the filter paper used in the step (2), heating and boiling the filter paper to remove water-soluble impurities, soaking the filter paper in a 1 mol/L EDTA-Na solution to complex metal ions in the paper base, soaking the filter paper in ethanol to remove organic impurities, modifying the filter paper with 0.5wt% of chitosan, and airing the modified filter paper for later use.
(4) Culture of cells on paper chip and detection of hydrogen peroxide
Placing the cell suspension in the paper chip prepared in step (3), placing the whole device in a culture dish, and placing the culture dish in 5% CO2The culture medium was changed at a place near water in the 37 ℃ incubator every two days. Fixing the paper chip and the modified silk-screen printing electrode on a self-made device, as shown in figure 1, the self-made device comprises a hydrogen peroxide detection box 1, the hydrogen peroxide detection box 1 is a square box, one side surface of the hydrogen peroxide detection box 1 is provided with a nitrogen inlet, the other side surface of the hydrogen peroxide detection box is provided with a nitrogen outlet, a middle cavity body can be used for placing the paper-based microfluidic chip manufactured in the step 2, a cell culture pond 2 is arranged in a hole of a porous PDMS layer 11 on the upper layer of the paper-based microfluidic chip, a PC12 cell 12 is arranged in the cell culture pond 2, a silk-screen printing carbon electrode 4 with a working electrode 7, a reference electrode 6 and a counter electrode 8 is inserted between a paper-based carrier 9 and a closed PDMS layer 10 in the paper-based microfluidic chip (or between the porous PDMS layer 11 and the paper-based carrier 9 on the upper layer, as long as the silk-screen printing carbon electrode 4 can contact with the cell to be detected, the PC12,stimulating cells to generate hydrogen peroxide by using a phorbol ester solution, setting parameters of an electrochemical workstation, setting parameters of the electrochemical workstation, and detecting the hydrogen peroxide in the cells by adopting a time current method.
Example 2:
a linear relation detection step of an electrochemical biosensor for online detection of hydrogen peroxide in cells based on combination of a paper chip and a screen-printed electrode, which is used for online monitoring of hydrogen peroxide released by PC12 cells, comprises the following steps:
when the growth state of PC12 cells (adrenal pheochromocytoma cells) is observed to be good after growing to 80% of the culture bottle, the culture solution is poured out, the cells are rinsed for three times by Phosphate Buffer Solution (PBS), an appropriate amount of PBS solution (0.1 mol/L, pH 7.4) is used for contacting the cells, the cells are placed in an incubator for culturing for 5 minutes, the PBS buffer solution contacting the PC12 cells is collected, hydrogen peroxide solutions with a series of concentrations are prepared, and the current response value of hydrogen peroxide with different concentrations at the voltage of-0.3V and the time of 50 s is recorded by adopting a time-current (i-t) method.
As can be seen from FIG. 3, the current response signal increased with increasing hydrogen peroxide concentration in the PBS buffer to which the cells were exposed. The hydrogen peroxide concentration is in a good linear relation with the current response value in the range of 0.1-8 mu mol/L, and the linear equation I (mu A) = 0.0538 + 0.1585CH2O2 (μmol/L)，R2 = 0.9944, detection limit 0.0232 μmol/L (S/N = 3).
Example 3:
a method for online monitoring an AD model by using an electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen-printed electrode comprises the following steps:
taking PC12 cells in logarithmic growth phase, digesting, centrifuging, re-suspending with complete culture solution, and counting to 5 × 104one/mL, planted in a paper-based chip, and divided into a control group, an Alzheimer Disease (AD) model group, and A beta25-35+ Cur group, the hydrogen peroxide released after phorbol ester stimulation of PC12 cells in each experimental group was monitored on-line by time-current (i-t) method, and the results are shown in FIG. 4.
As can be seen from FIG. 4, the channelsAβ25-35The current response value of the AD model group constructed by induction is larger than that of the control group, the content of the hydrogen peroxide released by stimulation is 224.30% of that of the control group, and after curcumin is dried, Abeta is obtained25-35The current response value of the + Cur group becomes smaller but larger than that of the control group, and the generated hydrogen peroxide content is 124.32% of that of the control group. Therefore, the expression level of the hydrogen peroxide in the AD model group cells is the highest, but the expression level of the hydrogen peroxide is obviously reduced after the curcumin is dried, namely the curcumin can improve the activity of the AD model cells.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principles of the present invention are intended to be included within the scope of the present invention.
Claims (6)
1. An electrochemical biosensor for online detection of hydrogen peroxide in cells based on a paper chip combined with a screen-printed electrode is characterized in that the paper chip takes filter paper as a carrier and a printing electrode modified by a nano platinum/multi-wall carbon nanotube material as a working electrode to form the biosensor for detecting hydrogen peroxide in cells by an electrochemical method; the paper chip is of a three-layer structure, the upper layer is a polydimethylsiloxane elastomer layer, and a round hole is formed in the middle of the polydimethylsiloxane elastomer layer and is used as a cell culture pool and a detection sample groove; the middle layer is a filter paper layer which is used as a carrier for culturing cells; the lower layer is a polydimethylsiloxane elastomer layer without a round hole and used as a carrier of the paper chip, and the three layers are tightly fixed by a clamp; the filter paper layer adopts the filter paper processed by the following method: heating and boiling filter paper to remove water-soluble impurities, soaking the filter paper in an EDTA-Na solution to complex metal ions in the paper base, soaking the filter paper in ethanol to remove organic impurities, modifying the filter paper with chitosan, and airing the filter paper for later use.
2. The electrochemical biosensor for on-line detection of hydrogen peroxide in cells based on paper chip combined with silk screen printed electrode as claimed in claim 1, wherein the hydrogen peroxide concentration is in the range of 0.1-8 μmol/L and the current response value is goodGood linearity, linear equation I (μ a) = 0.0538 + 0.1585CH2O2 (μmol/L)，R2 = 0.9944, detection limit 0.0232 μmol/L.
3. The method for detecting the hydrogen peroxide in the cells based on the electrochemical biosensor for detecting the hydrogen peroxide in the cells on line by combining the paper chip and the silk screen printed electrode as claimed in claim 1, which comprises the following steps:
(1) designing and manufacturing a paper chip: the paper chip is of a three-layer structure, the upper layer is a polydimethylsiloxane elastomer layer, and a round hole is formed in the middle of the polydimethylsiloxane elastomer layer and is used as a cell culture pool and a detection sample groove; the middle layer is a filter paper layer which is used as a carrier for culturing cells; the lower layer is a polydimethylsiloxane elastomer layer without a round hole and used as a carrier of the paper chip, and the three layers are tightly fixed by a clamp; the filter paper layer adopts the filter paper processed by the following method: heating and boiling filter paper to remove water-soluble impurities, then soaking the filter paper in an EDTA-Na solution to complex metal ions in the paper base, then soaking the filter paper in ethanol to remove organic impurities, finally modifying the filter paper with chitosan, and airing the filter paper for later use; (2) determination of hydrogen peroxide in cells: taking the cell suspension, placing the cell suspension in the paper chip prepared in the step (1), placing the paper chip in a culture dish, and placing the culture dish in 5wt% CO2Changing the culture solution in a 37 ℃ incubator in a place close to water every two days; after the culture is finished, a phorbol ester solution is used for stimulating cells to generate hydrogen peroxide, the paper chip and a screen printing electrode of the modified nano platinum/multi-wall carbon nanotube material are used as working electrodes to pass through an electrochemical workstation together, and the hydrogen peroxide in the cells is detected by adopting a time current method.
4. The method for detecting hydrogen peroxide in cells of the electrochemical biosensor based on the combination of the paper chip and the silk-screen printed electrode for on-line detection of hydrogen peroxide in cells as claimed in claim 3, wherein when the growth of PC12 cells reaches 80% of the growth state of the culture flask, the culture solution is poured off, and after the cells are rinsed with phosphate buffer solution, the cells are contacted with a proper amount of PBS solutionThe cell is placed in an incubator for culture, PBS buffer solution contacting with PC12 cells is collected, hydrogen peroxide solution with a series of concentrations is prepared by the solution, the hydrogen peroxide solution is placed on a paper chip and is used together with a screen-printed electrode of a modified nano platinum/multi-wall carbon nanotube material to record current response values of hydrogen peroxide with different concentrations and 50 s of time at the voltage of-0.3V, the current response signals are increased along with the increase of the hydrogen peroxide concentration in the PBS buffer solution contacting with the cells, the hydrogen peroxide concentration is in good linear relation with the current response values within the range of 0.1-8 mu mol/L, and the linear equation I (mu A) = 0.0538 + 0.1585CH2O2 (μmol/L)，R2 = 0.9944, detection limit 0.0232 μmol/L (S/N = 3).
5. The method for detecting the PC12 cells and the AD model cells by using the electrochemical biosensor for detecting the hydrogen peroxide in the cells on line based on the paper chip combined with the silk-screen printed electrode as claimed in any one of claims 1-2, wherein the paper chip is used as a PC12 cell culture and detection carrier, and the nano platinum/multi-walled carbon nanotube modified printing electrode is used as a working electrode to realize the on-line activity detection of the PC12 cells and the AD model cells.
6. The method of claim 5, wherein the PC12 cells and AD model cells are obtained from PC12 cells in logarithmic growth phase, digested, centrifuged, resuspended in complete culture medium, and counted to a concentration of 5X 104And each/mL is planted in a paper-based chip, and the online activity detection of PC12 cells and AD model cells is realized by using a time-current method by using a nano platinum/multi-wall carbon nanotube modified printing electrode as a working electrode.
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| CN110514714B (en) * | 2019-07-03 | 2021-10-08 | 福建医科大学 | Polylactic acid/paper hybrid chip for on-line monitoring dopamine release of nerve cells |
| CN112461751B (en) * | 2020-10-16 | 2022-04-26 | 江苏大学 | Cancer cell activity detection and evaluation device and method based on multi-adhesion strength fusion |
| CN113308937A (en) * | 2021-05-28 | 2021-08-27 | 陕西科技大学 | Cell culture paper and preparation method and application thereof |
| CN114965630A (en) * | 2022-06-06 | 2022-08-30 | 晨光生物科技集团股份有限公司 | Silk-screen printing electrode for detecting antioxidant activity and preparation and detection methods thereof |
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