Application of gold nanorod solution in primary screening reagent for exfoliated tumor cells

Abstract

The present invention relates to the application of a gold nanorod solution in a primary screening reagent for exfoliated tumor cells. The gold nanorod solution is etched by a redox reaction in a Gly-HCl buffer solution to quantify intracellular glutathione and glucose and obtain an optical photograph of the color of the etched gold nanorod solution. The obtained optical photograph is subjected to RGB color extraction and then judged to ultimately determine the cell type. The present invention utilizes glutathione overexpressed in tumor cells to reduce a potassium iodate solution to generate an etchant, etches gold nanorods, and simultaneously utilizes the highly absorbed glucose in tumor cells to react with the etchant to inhibit the progress of the etching reaction and produce a stable color change. Tumor cells are visually and accurately identified based on the color changes under two conditions, and are suitable for early prevention and diagnosis of cancer diseases.

Description

Technical Field

[0001] The invention belongs to the field of nano material science and detection technology, and relates to the application of a gold nanorod solution in an exfoliated cell primary screening reagent. Background Art

[0002] At present, exfoliative cytology requires cumbersome preparation steps and fresh samples to prevent cell death and changes in cell morphology. At the same time, cytology also has high requirements for testers and takes a long time to test. There is currently no method for quickly and accurately distinguishing most tumors from normal cells. Therefore, the development of an accurate and rapid initial screening kit for exfoliated tumor cells is of great significance for the rapid clinical detection of exfoliated tumor cells.

[0003] The Chinese patent document with publication number CN109540833A discloses a method for identifying in vitro tumor tissue based on terahertz time-domain spectroscopy and its application. It first proposed a method for identifying tumors using terahertz time-domain spectroscopy, using the specific absorption peak of the terahertz time-domain spectrum of tumor tissue to identify whether the tissue to be tested is non-tumor tissue, breast cancer tissue or rectal cancer tissue through comparison analysis. The Chinese patent document with publication number CN110229913A discloses a broad-spectrum tumor marker nucleic acid, which can be used for clinical tumor diagnosis, typing, detection and prognosis after PCR analysis. However, these methods have not yet been proven to be applicable to most tumor diagnoses and require expensive instruments, which is not conducive to on-site detection.

[0004] As one of the common sensing technologies, colorimetric sensors have been used to detect various targets through color changes visible to the naked eye or UV-visible spectra. Traditional colorimetric reporter probes are small organic molecules with chromophores, which are generally unstable in color development reactions and easily degraded during storage. In addition, these small organic molecule chromophores can usually only indicate the target through a single color change, which causes great errors in colorimetric detection by the naked eye. It is well known that the human eye is insensitive to changes in optical density (no spectral peak shift), but is sensitive to color changes (with spectral peak shift). Experts estimate that the human eye can distinguish up to 10 million colors. Therefore, it is foreseeable that if multiple colors are presented in colorimetric sensors, the accuracy of visual inspection can be significantly improved. In recent years, plasmonic sensors with multicolor changes (e.g., gold nanorods) have become a new research field. The main factor for the success of multicolor changes is that the LSPR changes of nanoparticles cover most of the visible absorption spectrum. LSPR is sensitive to changes in the size, shape, and dielectric environment of nanoparticles, so that etching or growth of nanoparticles leads to changes in plasmon bands, which in turn produce different colors.

[0005] Although the multiple colors generated by the changes in plasma bands have greatly improved the accuracy of colorimetric detection, false positive signals caused by factors such as the tester and the test environment are still inevitable. Based on the popularity of smartphones, colorimetric sensors are combined with smartphones. By using smartphone cameras and software analysis, colorimetric signals can be accurately and quickly detected, providing great potential for on-site sensing applications. Summary of the invention

[0006] The purpose of the present invention is to provide an application of a gold nanorod solution in a primary screening reagent for exfoliated cells. The highly expressed glutathione and glucose in tumor cells are visually detected and combined with a smart phone and a logic gate to achieve rapid and accurate differentiation of exfoliated tumor cells, which is suitable for clinical cancer diagnosis.

[0007] The present invention specifically comprises the following steps:

[0008] Step 1: removing impurities and lysing red blood cells from the exfoliated cells of the sampled cancer patients;

[0009] Step 2, enriching and washing the nucleated cells after the impurities and red blood cells are removed in step 1;

[0010] Step 3: repeatedly freeze-thaw and ultrasonically lyse the nucleated cell precipitate collected in step 2, and after centrifugation, take the supernatant to obtain a cell lysis solution;

[0011] Step 4: Add the cell lysis solution, potassium iodate solution and gold nanorod solution obtained in step 3 to a Gly-HCl buffer solution containing 40-50 mM sodium bromide and 8-10 mM hexadecyltrimethylammonium bromide. At this time, the glutathione and potassium iodate solution in the cell lysis solution undergo a redox reaction to etch the gold nanorod solution. Finally, the etched gold nanorod solution is photographed to obtain an optical photograph Img1.

[0012] The preparation method of the gold nanorod solution is as follows:

[0013] First, prepare the gold seed solution: dissolve 3.6445g of CTAB in 9.75mL of ultrapure water, then add 0.25mL of 0.01M HAuCl4 solution and stir for 3-5min at a speed of 400-700rpm, at which time the solution is uniformly yellow. Then, adjust the speed to 1200rpm and add 600mL of 0.01M NaBH4 solution prepared with ice water to the yellow solution and stir for 2 minutes, at which time the solution is brown, and obtain the gold seed solution; place the obtained brown gold seed solution in a 28℃ water bath for aging for 2h for use.

[0014] Then, prepare a gold nanorod solution: dissolve 18.225 g of CTAB in 500 mL of ultrapure water, add 25 mL of 0.01 M HAuCl4 solution, 5 mL of 0.01 M AgNO3 solution, 4 mL of 0.1 M ascorbic acid solution and 10 mL of 1 M HCl solution in sequence at a speed of 400-700 rpm and a water bath of 28-30 ° C, stir the mixture evenly, add 680-720 μL of the prepared gold seed solution, stir for 30-60 s, adjust the speed to zero, and let stand overnight in a water bath of 28-30 ° C to obtain a gold nanorod solution.

[0015] Finally, the obtained gold nanorod solution was centrifuged at 10,000 rpm for 3 times and redispersed with deionized water. The final concentration of the gold nanorod solution after centrifugation was 1 to 1.5 nM.

[0016] The gold nanorod solution is prepared by a classic seed-mediated method using chloroauric acid, silver nitrate, ascorbic acid, sodium borohydride, hydrochloric acid and hexadecyltrimethylammonium chloride as raw materials. The aspect ratio of the gold nanorod is 4:1-5:1, and the length is 60-80nm. The gold nanorods with corresponding parameters can produce colorful colors based on LSPR optical properties under etching conditions, and the visual detection sensitivity is high.

[0017] Step 5: The cell lysis solution obtained in step 3 is mixed with potassium iodate solution and then mixed with sodium hydroxide solution to perform an oxidation reaction in a dark environment to obtain a mixed solution to generate an etchant.

[0018] At this time, the glucose in the cell lysis solution reacts with the iodine element generated by the oxidation reaction, reducing the iodine content in the etchant and inhibiting the redox reaction of glutathione in the cell lysis solution and the potassium iodate solution, thereby inhibiting the etching of the gold nanorod solution.

[0019] Step six: Add the mixed solution (etchant) obtained in step five and the gold nanorod solution prepared in step four to a Gly-HCl buffer solution containing 40-50 mM sodium bromide and 8-10 mM hexadecyltrimethylammonium bromide, finally obtaining etched gold nanorod solution 2 and taking a photo to obtain an optical photo Img2.

[0020] Step 7. Input the optical photograph Img1 obtained in step 4 and the optical photograph Img2 obtained in step 6 into the RGB extraction program. When G1 / B1<0.9-0.94 and R2 / B2<1-1.1, that is, the concentration of glutathione is higher than 1 mM and the concentration of glucose is higher than 0.5 mM, the cell is determined to be a tumor cell.

[0021] Furthermore, the exfoliated cells collected from the patient in step 1 are not less than 10 3A membrane with a pore size of about 100 mm is installed on a syringe. The collected patient's exfoliated cells are filtered three times using the syringe, and then centrifuged at a centrifugal force of 500-700 g for 3-5 minutes. The upper layer of liquid is removed to remove impurities and enrich the cell particles, leaving the nucleated cell precipitate in the lower layer.

[0022] Preferably, the erythrocyte lysis is performed using an erythrocyte lysis solution, and the preparation method of 1L of erythrocyte lysis solution is as follows: weigh 1g of potassium bicarbonate, 8.3g of ammonium chloride and 0.037g of EDTA.2Na, dissolve in 500mL of ultrapure water, and finally adjust the pH value to 7.2-7.4 with KOH and HCl, and finally adjust the solution to 1L. The prepared erythrocyte lysis solution is sterilized at a pressure of 0.105MPa and a temperature of 121°C for 15-30min, and the sterilized erythrocyte lysis solution is stored at 4°C.

[0023] Furthermore, the washing of the nucleated cells in step 2 is performed using 0.01 M PBS buffer (pH=7.2-7.4). The nucleated cell precipitate is dispersed with PBS buffer, and then centrifuged at a centrifugal force of 500-700 g for 3-5 minutes. The process is repeated three times, and finally the precipitate is dispersed in PBS buffer.

[0024] Preferably, the enriched nucleated cells are dispersed in 1-2 mL of 0.01 M PBS buffer (pH = 7.2-7.4), ultrasonicated in an ultrasonic cleaner for 5-10 s, then centrifuged at 500-700 g for 5 min to remove the supernatant, the process is repeated three times, and the precipitate is dispersed in 10-100 mL of 0.01 M PBS buffer (pH = 7.2-7.4) for the last time.

[0025] Preferably, the repeated freezing and thawing and ultrasonic lysis in step three is to completely freeze the nucleated cells obtained in step two in a -20°C refrigerator and then slowly thaw them at room temperature, then ultrasonicate the lysed cells in an ultrasonic cleaner with a power of 100 W for 10 to 30 minutes, and repeat the freezing and thawing and ultrasonic lysis 5 to 6 times.

[0026] After the freeze-thaw and ultrasonic lysis were completed, the obtained mixed solution was centrifuged at 3000 rpm for 5 min, and the supernatant was taken to obtain a cell lysis solution.

[0027] Preferably, the volume ratio of the cell lysis solution, potassium iodate solution, gold nanorod solution and buffer solution in step 4 is 1:1:20:90, and the concentration of the potassium iodate solution is 50-55 mM. The etching reaction is carried out in a water bath at 45-55° C. for 6-7 minutes.

[0028] Preferably, in step 5, the volume ratio of the cell lysis solution, potassium iodate solution and sodium hydroxide solution is 4:1:4. The concentration of the sodium hydroxide solution is 20-25 mM.

[0029] The oxidation reaction in the dark environment is carried out at room temperature for 10 to 15 minutes.

[0030] Preferably, the volume ratio of the buffer solution, the mixed solution and the gold nanorod solution in step six is ​​90:9:2; the redox reaction and etching process are carried out at a temperature of 45 to 55° C., and the etching process is carried out for 6 to 7 minutes.

[0031] Preferably, the RGB extraction procedure described in step seven is:

[0032] import os

[0033] import cv2

[0034] import numpy as np

[0035] import math

[0036] path="C: / Users / Administrator / Desktop / rgb / picture"

[0037] file_name1="test1.png"

[0038] ##get

[0039] img1=cv2.imread(os.path.join(path,file_name1),1)

[0040] #mean1

[0041] r1 = np.mean(img1[:,:,2])

[0042] g1 = np.mean(img1[:,:,1])

[0043] b1 = np.mean(img1[:,:,0])

[0044] print(r1,g1,b1)

[0045] The present invention utilizes glutathione overexpressed in tumor cells to reduce potassium iodate solution to generate an etchant, and etches a gold nanorod solution to produce a stable color change. At the same time, glucose highly absorbed in tumor cells is used to react with the etchant to inhibit the etching reaction and produce a stable color change. Tumor cells can be visually identified within 30 minutes based on the color changes under two conditions, and the application prospect is broad in the early prevention and diagnosis of cancer diseases. The detached tumor cells are accurately distinguished by using an RGB extraction program and an "AND" logic gate judgment program. It has the advantages of being fast, low cost, and high color resolution, and can be used for the rapid and accurate distinction of detached tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is a TEM image of the gold nanorod solution prepared in Example 1, with a scale of 100 nm;

[0047] Figure 2 The ultraviolet spectrum of the gold nanorod solution prepared in Example 1;

[0048] Figure 3 The effect of different concentrations of glutathione on the longitudinal LSPR peak shift of gold nanorods and the optical photographs;

[0049] Figure 4 The UV spectra and optical photos of different concentrations of glucose inhibiting the longitudinal LSPR peak shift of gold nanorods;

[0050] Figure 5 To pass RGB Figure 3 and Figure 4 A logic gate analysis program established after extracting the optical photos in the image;

[0051] Figure 6 To visually distinguish 293T cells and MCF-7 cells using gold nanorod etching method. DETAILED DESCRIPTION

[0052] The present invention will be further described below in conjunction with the accompanying drawings and embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0053] The application of gold nanorod solution in the primary screening reagent of exfoliated cells is to etch the gold nanorod solution through redox reaction in Gly-HCl buffer solution to quantify the intracellular glutathione and glucose and obtain an optical photograph of the color of the etched gold nanorod solution. The obtained optical photograph is judged after RGB color extraction to finally determine the cell type. The glutathione overexpressed in tumor cells is used to reduce the potassium iodate solution to generate an etchant to etch the gold nanorods. At the same time, the highly absorbed glucose in the tumor cells reacts with the etchant to inhibit the etching reaction and produce a stable color change. The tumor cells are visually and accurately identified based on the color changes under the two conditions, which is suitable for the early prevention and diagnosis of cancer diseases.

[0054] First, prepare the gold nanorod solution, specifically:

[0055] Preparation of gold seed solution: Dissolve 3.6445g of CTAB in 9.75mL of ultrapure water, then add 0.25mL of 0.01M HAuCl4 solution and stir for 3-5min at a speed of 400-700rpm, at which time the solution is uniformly yellow. Then, adjust the speed to 1200rpm and add 600mL of 0.01M NaBH4 solution prepared with ice water to the yellow solution and stir for 2 minutes, at which time the solution is brown. Finally, place the brown gold seed solution in a 28-30℃ water bath for aging for 2h for use.

[0056] Preparation of gold nanorod solution: First, dissolve 18.225 g of CTAB in 500 mL of ultrapure water, add 25 mL of 0.01 M HAuCl4 solution, 5 mL of 0.01 M AgNO3 solution, 4 mL of 0.1 M ascorbic acid solution and 10 mL of 1 M HCl solution in sequence at a speed of 700 rpm and a water bath of 28 to 30 ° C, stir the mixture evenly, add 680 to 720 μL of the prepared gold seed solution, stir for 30 to 60 seconds, adjust the speed to 0 rpm, and let stand overnight in a water bath of 28 to 30 ° C to obtain a gold nanorod solution.

[0057] The aspect ratio of the obtained gold nanorods is 4:1 to 5:1, and the length is 60 to 80 nm. The TEM image of the gold nanorods is shown in FIG. Figure 1 As shown in the figure, the UV spectrum of gold nanorods is as follows Figure 2 As shown, the longitudinal LSPR peak of the gold nanorod is at 819 nm.

[0058] The obtained gold nanorod solution was centrifuged at 10000 rpm for 3 times and redispersed with deionized water. The final concentration of the gold nanorod solution after centrifugation was 1 to 1.5 nM.

[0059] The gold nanorod solution was etched with glutathione of different concentrations:

[0060] In a 700mL centrifuge tube, 225-230μL of Gly-HCl buffer solution (pH=2.8) containing 40-50mM sodium bromide and 8-10mM hexadecyltrimethylammonium bromide, 2.5-3μL of 50-55mM potassium iodate solution, and 2.5-3μL of glutathione solutions of different concentrations (0.27mM, 0.4mM, 0.6mM, 0.8mM, 1.2mM, 1.7mM, 2.5mM, 3.5mM, 5mM, 7mM, 10.5mM and 15mM) were added, and finally 50-60μL of the prepared centrifuged gold nanorod solution was added. After mixing on an oscillator for 30s, the mixture was placed in a water bath at 45-55℃ for 6-7min. The obtained solution was subjected to UV spectrum measurement and optical photographs were taken, and the optical photographs were named Img1.

[0061] Figure 3 A is the effect of different concentrations of glutathione on the longitudinal LSPR peak shift of gold nanorods; Figure 3 As can be seen in A, with the increase of glutathione concentration, the longitudinal LSPR peak of the gold nanorods gradually blue-shifted and finally only a single plasma peak of about 530 nm remained, indicating that the gold nanorods were finally etched into spherical nanoparticles. Figure 3 B is an optical photograph of glutathione etching gold nanorod solutions with different concentrations. The color of the gold nanorod solution changes from brown-red → gray → gray-green → green → blue-green → blue → purple → purple-red → pink as the glutathione concentration increases.

[0062] Different concentrations of glucose inhibit the etching of gold nanorods:

[0063] In a 700mL centrifuge tube, add 2.5-3μL of glutathione solution with concentrations of 0.4mM, 0.6mM, 0.8mM, 1mM, 3mM, 6mM, 9mM, 12mM and 15mM and 2.5-3μL of 50-55mM potassium iodate solution, mix them evenly on an oscillator and react in a 45-55℃ water bath for 10-15min. Then, add 10-12μL of 20-25mM sodium hydroxide solution and 10μL of 0.2mM, 0.3mM, 0.4mM, 0.5mM, 1mM, 1.5mM, 2mM, 2.5mM and 3mM glucose solution to each tube, mix on an oscillator for 30s, and react in the dark for 10-15min. Then, add 225-250 μL of Gly-HCl buffer solution (pH=2.8-3.0) containing 40-45 mM sodium bromide and 8-10 mM hexadecyltrimethylammonium bromide and 50-60 μL of the prepared centrifuged gold nanorod solution to each tube, mix on an oscillator for 30 seconds, and place in a 45-55°C water bath for 6-7 minutes.

[0064] Figure 4 A is the effect of different concentrations of glutathione and glucose on the longitudinal LSPR peak shift of gold nanorods; Figure 4 B is an optical photograph of gold nanorod solutions etched with glutathione and glucose at different concentrations, and the optical photograph is named Img2.

[0065] Using RGB extraction program Figure 3 Optical photographs of B Img1 and Figure 4 The three components of R, G and B were extracted from the optical photograph Img2 of B. Subsequently, the three independent components were combined in pairs, and the correlations of R / B, G / B and R / G with glutathione and glucose concentrations were compared, respectively. Figure 5 The logic gate program for judging cell types by optical photos is given in the figure. The pink judgment program is for judging Img1. When Img1 satisfies G1 / B1<0.94, the concentration of glutathione is higher than 1mM; the blue judgment program is for judging Img2. When Img1 satisfies R2 / B2<1.1, the concentration of glutathione is higher than 1mM and the concentration of glucose is higher than 0.5mM. It was finally determined that when Img1 satisfies G1 / B1<0.94 and Img2 satisfies R2 / B2<1.1, it can be used to judge that the detected cells are tumor cells.

[0066] In actual operation, the specific initial screening process is as follows:

[0067] Take out 100 μL of 293T and MCF-7 cell samples in a 2 mL centrifuge tube, put it in a -20 ° C refrigerator for 30 minutes to solidify, then thaw at room temperature, and then ultrasonicate in an ultrasonic cleaner for 10 minutes. Repeat the freeze-thaw and ultrasonic lysis 5 to 6 times. Finally, centrifuge at 3000 rpm for 5 minutes and take the supernatant. Dilute it with a PBS buffer solution with pH = 7.4 to obtain supernatants with different cell numbers to obtain the sample solution to be tested;

[0068] In a 700mL centrifuge tube, add 225μL of Gly-HCl buffer solution (pH=2.8) containing 40mM sodium bromide and 8mM hexadecyltrimethylammonium bromide, 2.5μL of 50mM potassium iodate solution, and 2.5μL of supernatant of 293T cells or MCF-7 cells with different cell numbers, and finally add 50μL of the centrifuged gold nanorod solution prepared in Example 1, mix on an oscillator for 30s, and place in a 55℃ water bath for 6 minutes and 25 seconds, and take optical photos of the obtained solution. Figure 6 The optical photos in A and 6C show the solution colors after glutathione etching of gold nanorods in 293T cells and MCF-7 cells with different cell numbers. Figure 6 The scatter plots in A and 6C are the relationship between the G1 / B1 value extracted from the optical photograph and the cell number. It can be seen that the G1 / B1 of 293T cells is greater than 0.94, while the G1 / B1 of MCF-7 cells is less than 0.94.

[0069] Add 2.5 μL of supernatant of 293T cells or MCF-7 cells with different cell numbers and 2.5 μL of 50 mM potassium iodate solution to a 700 mL centrifuge tube, mix the two evenly on an oscillator and react in a 55°C water bath for 10 min. Then add 10 μL of 20 mM sodium hydroxide solution to each tube and mix on an oscillator for 30 s, then react in the dark for 10 min. Then add 225 μL of Gly-HCl buffer solution (pH = 2.8) containing 40 mM sodium bromide and 8 mM hexadecyltrimethylammonium bromide and 50 μL of the centrifuged gold nanorod solution prepared in Example 1 to each tube, mix on an oscillator for 30 s, and place in a 55°C water bath for 6 minutes and 25 seconds. Figure 6 The optical photos in C and 6D show the solution colors after glutathione and glucose in 293T cells and MCF-7 cells with different cell numbers were simultaneously involved in the etching of gold nanorods. Figure 6 The scatter plots in A and 6C are the relationship between the R2 / B2 value extracted from the optical photograph and the number of cells. It can be seen that the R2 / B2 of 293T cells is greater than 1.1, while the R2 / B2 of MCF-7 cells is less than 1.1.

[0070] From the above data, it can be seen that the method of the present invention can quickly and accurately distinguish a wide range of exfoliated tumor cells through a logic gate program, indicating that the method of the present invention has broad application prospects in the early prevention and diagnosis of cancer diseases.

[0071] The embodiments described above provide a detailed description of the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, supplements or similar substitutions made within the scope of the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of gold nanorod solution in the preparation of exfoliated tumor cell primary screening reagent, the preparation method of the gold nanorod solution is as follows: First, prepare a gold seed solution: dissolve 3.6445 g of CTAB in 9.75 mL of ultrapure water, then add 0.25 mL of 0.01 M HAuCl4 solution and stir for 3-5 min at a speed of 400-700 rpm, at which time the solution is uniformly yellow; then, adjust the speed to 1200 rpm and add 600 mL of 0.01 M NaBH4 solution prepared with ice water to the yellow solution and stir for 2 minutes, at which time the solution is brown, and a gold seed solution is obtained; the obtained brown gold seed solution is placed in a 28 °C water bath for aging for 2 h for later use; Then, prepare the gold nanorod solution: dissolve 18.225 g of CTAB in 500 mL of ultrapure water, add 25 mL of 0.01 M HAuCl4 solution, 5 mL of 0.01 M AgNO3 solution, 4 mL of 0.1 M ascorbic acid solution and 10 mL of 1 M HCl solution in sequence at a speed of 400-700 rpm and a water bath of 28-30 °C, stir the mixture evenly, add 680-720 μL of the prepared gold seed solution, stir for 30-60 s, adjust the speed to zero and let stand overnight in a water bath of 28-30 °C to obtain the gold nanorod solution; Finally, the obtained gold nanorod solution was centrifuged at 10,000 rpm for three times and re-dispersed with deionized water. The final concentration of the gold nanorod solution after centrifugation was 1-1.5 nM; In a Gly-HCl buffer solution containing 40-50 mM sodium bromide and 8-10 mM hexadecyltrimethylammonium bromide, a detached tumor cell lysis solution, a potassium iodate solution and a gold nanorod solution were added. At this time, the glutathione in the cell lysis solution and the potassium iodate solution underwent a redox reaction to etch the gold nanorod solution. Finally, the etched gold nanorod solution was photographed together to obtain an optical photograph Img1; In a Gly-HCl buffer solution containing 40-50 mM sodium bromide and 8-10 mM hexadecyltrimethylammonium bromide, an etchant and a gold nanorod solution are added to finally obtain an etched gold nanorod solution II and take a photo to obtain an optical photo Img2; The obtained optical photographs Img1 and Img2 were input into an RGB extraction program. When G1 / B1<0.9~0.94 and R2 / B2<1~1.1, that is, the concentration of glutathione was higher than 1 mM and the concentration of glucose was higher than 0.5 mM, the cell was a tumor cell.

2. The use according to claim 1, characterized in that: The etching agent is: a cell lysis solution is mixed with a potassium iodate solution and then mixed with a sodium hydroxide solution to perform an oxidation reaction in a dark environment to obtain a mixed solution to generate an etching agent; At this time, the glucose in the cell lysis solution reacts with the iodine element generated by the oxidation reaction, reducing the iodine content in the etchant and inhibiting the redox reaction of glutathione in the cell lysis solution and the potassium iodate solution, thereby inhibiting the etching of the gold nanorod solution.

3. The use according to claim 1, characterized in that: The cell lysis solution is obtained by the following method: Step 1: removing impurities from the sampled exfoliated cells and lysing red blood cells; Step 2, enriching and washing the nucleated cells after the impurities and red blood cells are removed in step 1; Step 3: Repeatedly freeze-thaw and ultrasonically lyse the nucleated cell precipitate collected in step 2, and after centrifugation, take the supernatant to obtain a cell lysis solution.

4. The use according to claim 3, characterized in that: The second step is to filter with a membrane having a pore size of about 100 mm, and then centrifuge at a centrifugal force of 500-700 g for 3-5 min, remove the upper liquid to remove impurities and enrich the cell particles, leaving the nucleated cell precipitate in the lower layer; disperse the nucleated cell precipitate with PBS buffer, and then centrifuge at a centrifugal force of 500-700 g for 3-5 min, repeat the process three times, and finally disperse the precipitate in PBS buffer.

5. The use according to claim 3, characterized in that: The repeated freezing and thawing and ultrasonic lysis described in step three is that the nucleated cells obtained in step two are completely frozen in a -20 °C refrigerator and then slowly thawed at room temperature, and then the lysed cells are ultrasonicated in an ultrasonic cleaner with a power of 100 W for 10 to 30 min, and the above freezing and thawing and ultrasonic lysis are repeated 5 to 6 times; after the freezing and thawing and ultrasonic lysis are completed, the obtained mixed solution is centrifuged at 3000 rpm for 5 min, and the supernatant is taken to obtain a cell lysis solution.

6. The use according to claim 1, characterized in that: The volume ratio of the cell lysis solution, potassium iodate solution, gold nanorod solution and Gly-HCl buffer solution is 1:1:20:

90.

7. The use according to claim 2, characterized in that: The redox reaction generates an etchant to regulate the etching process of the gold nanorods at 45-55 °C for 6-7 min.

8. The use according to claim 2, characterized in that: The volume ratio of the cell lysis solution, potassium iodate solution and sodium hydroxide solution is 4:1:

4.

9. The use according to claim 2, characterized in that: The oxidation reaction time in the dark environment is 10 to 15 minutes.

10. The use according to claim 1, characterized in that: The volume ratio of the Gly-HCl buffer solution, the etchant and the gold nanorod solution is 90:9:2.

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