CN115598086B - Terahertz metamaterial biosensor for evaluating postoperative curative effect of glioma and application - Google Patents

Abstract

The invention belongs to the field of biomedical engineering, and relates to a terahertz metamaterial biosensor for evaluating the postoperative curative effect of glioma and application thereof. The terahertz metamaterial chip is arranged in a container for culturing cells; the terahertz metamaterial chip is a gold film in an array shape formed by a plurality of sensing units, and each row of sensing units are connected in series through connecting conductors; the sensing unit is of an inductance-capacitance resonator structure, a detection channel is formed between two electrodes of the inductance-capacitance resonator structure, and the detection channel is in a zigzag shape. The terahertz metamaterial biosensor provided by the invention can be used for rapidly, in-situ, label-free and destructive-free detection of proliferation inhibition and apoptosis promotion capability of tumor treatment simultaneously, so that the curative effect of auxiliary treatment is evaluated, and the establishment of an individual treatment scheme is guided.

Description

Terahertz metamaterial biosensor and its application for evaluating the efficacy of glioma surgery

Technical Field

The present invention belongs to the field of biomedical engineering and relates to a terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery and its application.

Background Art

The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

Due to the high heterogeneity and invasiveness of glioma cells, individuals have different sensitivities to the same adjuvant therapy (radiotherapy, chemotherapy, etc.), resulting in unsatisfactory effects of adjuvant therapy. Therefore, there is an urgent need for rapid and accurate detection technologies and methods to evaluate the efficacy of adjuvant therapy in order to guide individualized treatment.

Proliferation and apoptosis are the two most important tumor biological characteristics for evaluating the efficacy of adjuvant therapy. Proliferation is the division process of living cells, and the state does not change much, while apoptosis is a programmed cell death with phenomena such as cell atrophy and chromatin condensation. Therefore, cell apoptosis not only changes the number of living cells, but also changes the dielectric constant of tumor cells. According to the inventor's research, it is difficult to detect the proliferation and apoptosis of tumor cells simultaneously and quickly and accurately with existing technologies.

Summary of the invention

In order to address the deficiencies of the prior art, the purpose of the present invention is to provide a terahertz metamaterial biosensor and application for evaluating the postoperative efficacy of glioma. The terahertz metamaterial biosensor provided by the present invention can rapidly, in situ, label-free and non-destructively simultaneously evaluate the proliferation inhibition and apoptosis promotion abilities of tumor treatment, thereby guiding the formulation of individualized auxiliary treatment plans.

In order to achieve the above object, the technical solution of the present invention is:

On the one hand, a terahertz metamaterial biosensor for evaluating the postoperative efficacy of glioma comprises a terahertz metamaterial chip, wherein the terahertz metamaterial chip is arranged in a container for culturing cells; the terahertz metamaterial chip is a gold film in an array shape formed by a plurality of sensing units, wherein each row of sensing units is connected in series through a connecting conductor; the sensing unit is an inductor-capacitor resonator structure, wherein a detection channel is formed between two electrodes of the inductor-capacitor resonator structure, and the detection channel is zigzag.

The present invention uses gold to prepare a terahertz metamaterial chip, which can make the formed biosensor have better biocompatibility and low loss characteristics at terahertz frequencies. At the resonant frequency, the electric field is highly confined between the capacitor electrodes, so this area is a region that can respond sensitively to slight changes in the external environment. When the biosensor detects glioma cells, its resonant frequency will change with the changes in the number and state of the cells, thereby realizing the detection of the number and state of the cells. The present invention increases the effective area of the sensitive area by setting a tortuous detection channel, thereby increasing the stability of the detection.

On the other hand, a method for preparing the above-mentioned terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery includes the following process of preparing a terahertz metamaterial chip:

A photoresist layer is coated on the surface of the substrate, and a mask engraved with the same pattern as the terahertz metamaterial chip is covered on the surface of the photoresist layer. UV lithography is then performed, and then a layer of gold film is plated on the template after UV lithography by electron beam evaporation, and the excess photoresist is removed.

Thirdly, the application of the above-mentioned terahertz metamaterial biosensor for evaluating the postoperative efficacy of glioma in detecting the apoptosis rate of glioma cells.

Fourthly, a terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery is used to simultaneously monitor the proliferation and apoptosis of glioma cells.

A fifth aspect is the application of the above-mentioned terahertz metamaterial biosensor for evaluating the postoperative efficacy of glioma in evaluating the efficacy of adjuvant therapy.

The beneficial effects of the present invention are:

The metamaterial biosensor designed with interdigital capacitors in the present invention can effectively improve the area ratio, and its resonant frequency is very sensitive to changes in the dielectric constant caused by changes in the number and state of cells. Experiments have shown that a fixed number of glioma cells composed of various living cells and apoptotic cells show different responses to terahertz metamaterial biosensors, revealing different refractive indices in different cell states. Finally, under various therapies, combined with basic biological experiments, different numbers of glioma cells were characterized in situ. The relationship between the number of living cells, apoptotic cells and frequency shift was determined, which correspondingly revealed for the first time the relationship between tumor cell proliferation, apoptosis rate and frequency shift. Therefore, the present invention can be used to quickly, in situ, label-free and non-destructively evaluate the proliferation inhibition and apoptosis promotion capabilities of tumor treatments, thereby guiding personalized treatment of gliomas.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

FIG1 is a structure of a sensing unit of a terahertz metamaterial chip according to an embodiment of the present invention;

FIG2 is a schematic diagram of the manufacturing process of a terahertz metamaterial chip in an embodiment of the present invention, I. Spin coating, spin coating photoresist on a clean quartz substrate for 90 seconds at a speed of 4000 rpm, II. Ultraviolet lithography technology, transfer of metamaterial pattern from the mask to the substrate, III. Development, the transferred metamaterial pattern will appear in the developer, IV. Evaporation, evaporation of gold thin film by electron beam evaporation;

FIG3 is a characterization diagram of a terahertz metamaterial chip in an embodiment of the present invention, a, an image of a prepared biosensor, b, a metamaterial biosensor with an integrated polydimethylsiloxane (PDMS) cell culture chamber, c, a bare terahertz metamaterial chip, d, a terahertz metamaterial chip with glioma cells cultured in a PDMS cavity, and e, a terahertz metamaterial chip with glioma cells cultured in a PDMS cavity;

Figure 4, a, schematic diagram of THz-FDS detection of glioma cells with different apoptosis rates, b, c, THz spectra of biosensor and 3×10 ⁴ U251 cells after culturing with (b) TMZ and (c) IR for 48h to detect cell apoptosis, d, e, representative images and apoptosis rates of glioma cells detected by flow cytometry after glioma cells were treated with (d) 0, 400, 800, 1600μM TMZ and (e) 0, 6, 9, 12Gy IR for 48h, f, g, EdU cell proliferation assay results and EdU positive cell % of U251 glioma cells after (f) 48h TMZ and (g) 48h IR treatment. EdU positive cell % qualitatively predicts proliferation rate, scale bar, 200 μm, data are expressed as mean ± SD (n = 3, **p <0.01; ***p <0.001; ****p <0.0001);

FIG5 is a schematic diagram of using the proposed terahertz metamaterial biosensor to detect glioma cells under treatment;

FIG6 is a graph showing the characterization results of the proliferation experiment in an embodiment of the present invention, a, images of different numbers of U251 cells after culture on the biosensor (the initial number is n ₀ , and the number after culture is n _c ), b, terahertz spectrum of the quantitative variable U251 cell biosensor, c, relationship between Δf and the number of cells n ₀ and n _c ;

FIG7 is a schematic diagram of the process of treating glioma cells using different auxiliary treatment methods according to an embodiment of the present invention;

FIG8 is a graph showing the detection results of proliferation and apoptosis in an embodiment of the present invention, ab, the number of live and apoptotic cells of U251 glioma under treatment with 0, 400, 800 and 1600 μM TMZ (a) and irradiation with 0, 6, 9 and 12 Gy IR (b) according to formula (4), n ₀ =3×10 ⁴ U251 cells, c, determination of the terahertz transmittance of U251 cells under the action of TMZ, d, the relationship between Δf apoptotic cells and the number of live cells under TMZ treatment, e, the relationship between Δf and the proliferation rate and apoptosis rate under TMZ treatment, f, measurement of the terahertz transmittance of U251 cells under infrared irradiation, g, the relationship between Δf apoptotic cells and the number of live cells under IR treatment, h, the fitting relationship between UΔf and the proliferation rate and apoptosis rate under IR, i and j are 1600 μM TMZ, 120 μM LOM, 2 μM ETO (IC50 dose) (i) and 6 Gy IR, respectively. Δf of U251, LN229, and A172 glioma cell lines treated with IR (j) for 48 h;

DETAILED DESCRIPTION

It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and/or "including" are used in this specification, it indicates the presence of features, steps, operations, devices, components and/or combinations thereof.

In view of the fact that it is difficult to simultaneously and quickly and accurately detect the proliferation and apoptosis of tumor cells with the existing technology, and thus detect the sensitivity of glioma cells to adjuvant therapy, the present invention proposes a terahertz metamaterial biosensor and its application for evaluating the efficacy of glioma surgery.

A typical embodiment of the present invention provides a terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery, including a terahertz metamaterial chip, which is arranged in a container for culturing cells; the terahertz metamaterial chip is a gold film in an array shape formed by a plurality of sensing units, and each row of sensing units is connected in series through a connecting conductor; the sensing unit is an inductor-capacitor resonator structure, and a detection channel is formed between two electrodes of the inductor-capacitor resonator structure, and the detection channel is zigzag. Studies have shown that the terahertz metamaterial biosensor provided by the present invention can determine the relationship between the number of living cells, apoptotic cells and frequency shift, so that the proliferation inhibition and apoptosis promotion ability of tumor treatment can be evaluated quickly, in situ, label-free and non-destructively, and has good consistency with basic biological experiments.

The zigzag shape described in the present invention is, for example, an m-shape, a z-shape, a serpentine shape, a wave shape, etc. The embodiment of the present invention uses an m-shape detection channel for verification, and the effect is good.

In some embodiments, the terahertz metamaterial chip is disposed on a surface of a quartz substrate. The use of a quartz substrate can ensure the biocompatibility and low-loss characteristics of the biosensor.

In some embodiments, the container includes a culture chamber, the terahertz metamaterial chip serves as the bottom of the culture chamber, and the material of the culture chamber is polydimethylsiloxane (PDMS). The culture chamber made of PDMS can further improve the uniformity of cell growth and distribution.

In some embodiments, there are 150 to 250 sensing units per square millimeter of the terahertz metamaterial biosensor.

In some embodiments, the resonant frequency is 370-380 GHz.

In some embodiments, the thickness of the gold film is 80-120 nm.

Another embodiment of the present invention provides a method for preparing the above-mentioned terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery, including the following process of preparing a terahertz metamaterial chip:

A photoresist layer is coated on the surface of the substrate, and a mask engraved with the same pattern as the terahertz metamaterial chip is covered on the surface of the photoresist layer. UV lithography is then performed, and then a layer of gold film is plated on the template after UV lithography by electron beam evaporation, and the excess photoresist is removed.

In some embodiments, the process includes assembling the terahertz metamaterial chip and the container, placing the terahertz metamaterial chip on the upper surface of the bottom of the container, and then bonding the side wall of the container to the bottom of the container.

A third embodiment of the present invention provides an application of the above-mentioned terahertz metamaterial biosensor for evaluating the postoperative efficacy of glioma in detecting the apoptosis rate of glioma cells.

For example, the application in preparing or constructing a system for detecting the apoptosis rate of glioma cells.

A fourth embodiment of the present invention provides an application of the above-mentioned terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery in simultaneously monitoring the proliferation and apoptosis of glioma cells.

For example, the application in preparing or constructing a system for simultaneously monitoring the proliferation and apoptosis of glioma cells.

A fifth embodiment of the present invention provides an application of the above-mentioned terahertz metamaterial biosensor for evaluating the postoperative efficacy of glioma in evaluating the efficacy of adjuvant therapy.

For example, the application in the preparation or construction of a system for evaluating the efficacy of adjuvant therapy.

The evaluation of the efficacy of adjuvant therapy is preferably the evaluation of the efficacy of adjuvant therapy for glioma.

The application of the present invention may be for the purpose of diagnosis and treatment of diseases or for the purpose of diagnosis and treatment of non-diseases. For the purpose of diagnosis and treatment of non-diseases, for example, scientific research on the proliferation and apoptosis of glioma cells.

In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

Example

Design of Terahertz Metamaterial Biosensor:

The terahertz metamaterial chip of this embodiment is composed of periodic inductive and capacitive coupled resonators. The designed metamaterial biosensor is simulated and optimized using HFSS and finite element method. Periodic boundaries, namely perfect electric conductors (PECs) and perfect magnetic conductors (PMCs), are used for simulation.

The sensing unit of the terahertz metamaterial chip is shown in Figure 1a.

The production process of terahertz metamaterial chip is shown in Figure 2.

First, the quartz substrate was cleaned with Decon solution, deionized water, acetone, and ethanol and dried with a nitrogen gun; then the clean and dry quartz was transferred and fixed on a spin coater, and AR-P with a thickness of about 1 μm was spin-coated on the surface. 5350 photoresist layer; place the quartz wafer coated with photoresist on a hot plate and bake at 110℃ for 3 minutes until the photoresist is completely dried; fix the mask plate engraved with the same pattern as the terahertz metamaterial chip and the dried quartz wafer to the UV lithography machine, and after 7 seconds of UV lithography, the required pattern is formed on the photoresist layer of the quartz wafer; remove the quartz wafer from the UV lithography machine, put it into 5350 developer and soak it for 20 seconds, take it out and soak it in deionized water for 1 minute to remove the residual developer on the surface, then blow it dry with a nitrogen gun, and observe whether the morphology is complete under a microscope; then use electron beam evaporation to plate a 100nm thick gold film on the quartz wafer with complete morphology; use acetone to strip off the remaining photoresist to obtain a complete metamaterial metal pattern, and the device is prepared.

PDMS cavity preparation:

PDMS solution and curing agent were purchased from Dow Corning. PDMS solution and curing agent were mixed at a mass ratio of 10:1. Then, bubbles were removed in a vacuum oven for 30 minutes. Before pouring the mixed solution into the mold, a release agent was sprayed on the mold surface to facilitate the removal of the PDMS cavity. After baking on a hot plate at 70 °C for 120 minutes, the PDMS cavity was removed from the mold.

The terahertz metamaterial chip and the PDMS cavity are assembled as shown in Figure 3b.

Terahertz detection:

Metamaterial chips with or without glioma cells were characterized using Toptica THz-FDS TeraScan 1550, as shown in Figure 7. Before THz testing, the MTMs were rinsed with phosphate-buffered saline (PBS) for 90 seconds and then gently blown dry under a stream of nitrogen for 60 seconds. The label-free test took 5 minutes for each sample.

Characterization results:

Terahertz metamaterial biosensors:

Figure 5 shows the detection of glioma cells using the biosensor. It can be seen that the resonant frequency of the metamaterial biosensor changes with the change of cell number and state. Generally, the resonant frequency of the inductive-capacitive coupled resonator based on parallel plate capacitors is

where L and C are the equivalent inductance and capacitance, respectively. At the resonant frequency, the electric field is highly confined between the capacitor electrodes to achieve high sensitivity. In theory, C is sensitive to the dielectric constant ε between the capacitor electrodes and manipulates the resonant frequency f ₀ according to formula (1), which is the key physical mechanism of the proposed biosensor.

Figure 3a shows a biosensor containing several metamaterial units. To further improve the uniformity of cell growth and distribution, a culture chamber with polydimethylsiloxane (PDMS) was designed and integrated into the MTM biosensor, as shown in Figure 3b. Figures 3c-e are microscope images of the metamaterial chip, with cells cultured on the surface of the metamaterial chip with and without the PDMS chamber. It is clear that the PDMS chamber makes the glioma cells more evenly distributed.

Proliferation assay:

The proliferation and apoptosis rates of glioma cells under 48 h adjuvant therapy were defined as η _p and η _{a ,} respectively. 48 h is the recommended time for standard bioassays in "Biological Materials and Experimental Methods". Therefore, the number of viable cells and apoptotic cells after culture can be estimated as

n _l =n ₀ ×(1+n _p -n _a ) and n _a =n ₀ ×n _a , (4)

_n0 is the initial cell amount. In the absence of adjuvant therapy, _ηp is approximately a fixed value, which is suppressed in the case of treatment. In the absence of treatment, _ηa is usually a few percent, which is significantly improved in the case of treatment. This embodiment takes the U251 cell line as an example to study the detection of glioma cells by THz MTMs. The total cell number _nc and the apoptosis rate _ηa can be obtained by cell counting and flow cytometry, respectively. The proliferation rate _ηp is obtained by _nc and _ηa according to formula (4), and is verified by the relative proliferation rate obtained by the EdU cell proliferation test.

First, a 200-μl suspension of U251 cells with different concentrations was placed on a biosensor with a PDMS cavity and cultured for 8 h, and its distribution is shown in Figure 6a. Then, the cell biosensor was characterized by THz-FDS. Figure 6b describes the characteristic transmittance for different initial numbers n ₀ , and Figure 6c describes the relationship between Δf and the number of cells. As the number increases from 0 to 6×10 ⁴ cell, the resonance frequency gradually shifts to low frequencies. Considering the proliferation and apoptosis of glioma cells within 8 h, the proliferation and apoptosis rates of glioma cells after 48 h were determined by standard biological methods under the same culture conditions. The number of viable cells was estimated to be n _l =n ₀ ×(1+η _p -ηa) ^(8/48) , and n _c was approximately equal to n _l , because the apoptosis rate after 8 h was negligible (less than 0.7%). In this case, the quasi-linear formulas for Δf and cell number can be fitted as Δf = 14.21×n ₀ and Δf = 12.36×n _l , respectively. The experimental data of the other two glioma cell lines are shown in Table 1 , showing the same trend.

The fitting relationships with different slopes reflect the cell volume and proliferation rate of different glioma cell lines to a certain extent. In the relationship between Δf and n _c , the slope of the A172 cell line is greater than that of LN229, indicating that A172 glioma cells are larger and proliferate faster than LN229 cells.

Table 1 Detection of untreated glioma cells cultured for 8 hours by terahertz metamaterial biosensor

Apoptosis detection:

To investigate the effect of apoptotic cells, four groups of 3×10 ⁴ U251 cells cultured under different treatment conditions, consisting of different numbers of live and apoptotic cells, were tested using the MTM biosensor. As shown in Figure 4a, the cells digested from the culture plate after 48 h of treatment were normalized to obtain 3×10 ⁴ cells, which were cultured on the biosensor for another 5 h to obtain good adhesion and then characterized by THz-FDS. The transmittance characteristics are shown in Figure 4b, c. The apoptosis rate under TMZ and radiotherapy treatments was measured by flow cytometry, as shown in Figure 4d, e, respectively. Due to the increase in treatment dose, Δf gradually decreased as cell apoptosis increased. This indicates that the refractive index of apoptotic cells is smaller than that of live cells. However, the reduction in the number of live cells also leads to a smaller Δf, which means that Δf cannot be directly related to n _a . In addition, during the digestion and replantation process, apoptotic cells may collapse and live cells may proliferate, which introduces additional errors to the method.

Proliferation and apoptosis detection:

As shown in Figure 7, 3×10 ⁴ glioma cells were cultured on the biosensor for 48 hours and then directly measured by THz-FDS. As a reference group, 3×10 ⁴ (b ₀ ) glioma cells were cultured in a 48-well plate under the same treatment conditions, and the total number of tumor cells (n _c ) was calculated using a cell counter. η _p can be calculated according to formula (4) obtained in the previous step. The calculated n _l , n _a , η _p , and η _a are shown in Figure 8a, b and Table 3, showing good proliferation inhibition and apoptosis promotion capabilities. The calculated proliferation rate is consistent with the trend of the EdU cell proliferation experiment results in Figure 4f, g. The characteristic transmittance of U251 cells under TMZ and IR treatment is shown in Figure 8c, f. It can be seen that with the increase of the treatment dose, the number of live cells decreases, the number of apoptotic cells increases, and the number of total cells decreases, and its Δf shows a downward trend, as shown in Figure 8a, b. According to cell counting and biological experiments, the relationship between Δf and n _l and n _a under different treatments is plotted in Figure 8d, g. The fitting formula of the relationship curve can be obtained as Δf = A ₁ × n _l + A ₂ × n _a + A ₃ , where A ₁ , A ₂ and A ₃ are constants. The relationship between Δf and η _p and η _a can be fitted as Δf = B ₁ × η _p + B ₂ × η _a + B ₂ , where B ₁ , B ₂ and B ₃ are also constants (Figure 8e, h). Therefore, as long as Δf is characterized by terahertz MTMs, η _p and η _a can be fitted with it. Similar phenomena were also found for other glioma cell lines under various adjuvant therapies, as shown in Tables 3 and 4. This is the first time that THz MTMs have been shown to simultaneously detect apoptosis and proliferation rates of tumor cells.

Table 2 THz MTMs detection of glioma cells cultured for 48 h under different adjuvant treatments

Table 3 Relationship between Δf, n _l and n _a of three cell lines under different adjuvant treatments

Δf: resonance frequency shift; n _l : number of living glioma cells; _na : number of apoptotic glioma cells.

Table 4 Relationship between Δf, _ηp and _ηa of three cell lines under different adjuvant treatments

Δf: resonance frequency shift; n _l : number of living glioma cells; _na : number of apoptotic glioma cells.

In summary, the frequency shift of terahertz MTMs induced by living cells and apoptotic cells under treatment can be used to evaluate the efficacy of adjuvant therapy in situ, accurately, quickly and label-free. The smaller the Δf, the better the efficacy. The corresponding Δf of U251, LN229 and A172 cells at the IC50 doses of three drugs and the commonly used radiotherapy doses in clinical practice are shown in Figure 8i,j. Note that Δf is not just an efficacy indicator of a single biological indicator (proliferation or apoptosis), but an efficacy indicator of the combined response. Without adding any labels, each experiment takes only 5 minutes, which is 24 times shorter than flow cytometry and 60 times shorter than EdU cell proliferation method. Therefore, this method can well detect the proliferation and apoptosis characteristics of tumor cells under various treatment methods, and has the potential to guide surgeons to develop personalized treatment plans for glioma patients.

Conclusion, the present invention proposes a new terahertz metamaterial method to simultaneously monitor the proliferation and apoptosis of glioma cells and evaluate the efficacy of adjuvant therapy. First, the present invention proves that the characteristic frequency shift Δf is a comprehensive response to the change in the number and state of glioma cells. Then, through biological experiments, the relationship between the number of living cells and apoptotic cells and Δf is fitted, and the relationship between the proliferation rate and apoptosis rate and Δf is revealed. Under treatment conditions, the relationship between Δf, n _l and n _a is fitted as Δf = A ₁ × n _l + A ₂ × n _a + A ₃ . Correspondingly, the relationship between Δf and η _p and η _a can be fitted as Δf = B ₁ × η _p + B ₂ × η _a + B ₃ . Therefore, this method can simultaneously monitor the proliferation and apoptosis of glioma cells, and accurately, in situ, label-free, non-ionizing radiation, and quickly evaluate the sensitivity of glioma cells to adjuvant therapy.

The frequency shift method proposed by the present invention is non-specific and rapid in detecting the therapeutic efficacy.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims (4)

1. Application of a terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery in simultaneously monitoring the proliferation and apoptosis of glioma cells, characterized in that: The terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery comprises a terahertz metamaterial chip, which is arranged in a container for culturing cells; the terahertz metamaterial chip is a gold film in an array shape formed by a plurality of sensing units, and each row of sensing units is connected in series through a connecting conductor; the sensing unit is an inductor-capacitor resonator structure, and a detection channel is formed between two electrodes of the inductor-capacitor resonator structure; The detection channel is m-shaped; The terahertz metamaterial chip is arranged on the surface of a quartz substrate; The container comprises a culture chamber, the terahertz metamaterial chip serves as the bottom of the culture chamber, and the material of the culture chamber is PDMS; and , The fitting formula of the relationship curve ,in , and is a constant; :Resonance frequency shift; :Number of viable glioma cells; : Number of apoptotic glioma cells; and and The relationship curve fitting is ,in , and is a constant; :Resonance frequency shift; : Glioma cell proliferation rate; : Apoptosis rate of glioma cells. 2. A use as claimed in claim 1, characterized in that the thickness of the gold film is 80-120 nm. 3. An application as claimed in claim 1, characterized in that the method for preparing the terahertz metamaterial biosensor for evaluating the efficacy of glioma surgery comprises the following process of preparing the terahertz metamaterial chip: A photoresist layer is coated on the surface of the substrate, and a mask engraved with the same pattern as the terahertz metamaterial chip is covered on the surface of the photoresist layer. UV lithography is then performed, and then a layer of gold film is plated on the template after UV lithography by electron beam evaporation, and the excess photoresist is removed. 4. An application as claimed in claim 3, characterized in that it includes a process of assembling the terahertz metamaterial chip and the container, placing the terahertz metamaterial chip on the upper surface of the bottom of the container, and then bonding the side wall of the container to the bottom of the container.