CN115144519A - Single cell sample fingerprint detection method based on inorganic nanoparticles and application - Google Patents
Single cell sample fingerprint detection method based on inorganic nanoparticles and application Download PDFInfo
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- 239000002105 nanoparticle Substances 0.000 title claims abstract description 52
- 238000001514 detection method Methods 0.000 title claims abstract description 46
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 claims abstract description 61
- 239000011159 matrix material Substances 0.000 claims abstract description 28
- 238000004458 analytical method Methods 0.000 claims abstract description 14
- 238000004949 mass spectrometry Methods 0.000 claims abstract description 14
- NDLPOXTZKUMGOV-UHFFFAOYSA-N oxo(oxoferriooxy)iron hydrate Chemical compound O.O=[Fe]O[Fe]=O NDLPOXTZKUMGOV-UHFFFAOYSA-N 0.000 claims abstract description 5
- 238000005507 spraying Methods 0.000 claims abstract description 5
- 238000000816 matrix-assisted laser desorption--ionisation Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims abstract description 4
- 238000001869 matrix assisted laser desorption--ionisation mass spectrum Methods 0.000 claims abstract description 3
- 230000002503 metabolic effect Effects 0.000 claims description 13
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 11
- 239000008367 deionised water Substances 0.000 claims description 10
- 229910021641 deionized water Inorganic materials 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
- 239000004809 Teflon Substances 0.000 claims description 4
- 229920006362 Teflon® Polymers 0.000 claims description 4
- 150000001413 amino acids Chemical class 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 150000002500 ions Chemical class 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000001632 sodium acetate Substances 0.000 claims description 4
- 235000017281 sodium acetate Nutrition 0.000 claims description 4
- 239000001509 sodium citrate Substances 0.000 claims description 4
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 2
- 150000001720 carbohydrates Chemical class 0.000 claims description 2
- 230000009089 cytolysis Effects 0.000 claims description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 2
- 238000005464 sample preparation method Methods 0.000 claims 1
- 238000011161 development Methods 0.000 abstract description 2
- 230000004907 flux Effects 0.000 abstract description 2
- 230000003950 pathogenic mechanism Effects 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 230000005789 organism growth Effects 0.000 abstract 1
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- 210000004027 cell Anatomy 0.000 description 49
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 11
- 238000001819 mass spectrum Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 4
- 239000002207 metabolite Substances 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 3
- 239000013592 cell lysate Substances 0.000 description 2
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- 230000003111 delayed effect Effects 0.000 description 2
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- 238000003384 imaging method Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000002705 metabolomic analysis Methods 0.000 description 2
- 230000001431 metabolomic effect Effects 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 2
- 229940038773 trisodium citrate Drugs 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241000269370 Xenopus <genus> Species 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 210000002569 neuron Anatomy 0.000 description 1
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
Abstract
The invention discloses a single cell sample fingerprint detection method based on inorganic nanoparticles and application thereof, and relates to the technical field of analysis and detection. The detection method comprises the following steps: step 1: preparing an iron oxide micro-nano particle matrix; step 2: spraying the ferric oxide micro-nano particle matrix into a single cell sample containing biological micromolecules to prepare a sample to be analyzed; and step 3: performing MALDI mass spectrometry detection on the fingerprint of the sample to be analyzed; and 4, step 4: and analyzing the detection result of the MALDI mass spectrum to obtain a conclusion. The detection method of the invention has high sensitivity, low cost and high detection flux, meets the requirement of acquiring the high-flux fingerprint of the metabolome at the single cell level, can reveal the heterogeneity among cells, and has great application potential in deep interpretation of single cell level in the processes of organism growth and development, pathogenic mechanism and the like.
Description
Technical Field
The invention relates to the technical field of analysis and detection, in particular to a single cell sample fingerprint spectrum detection method based on inorganic nanoparticles and application thereof.
Background
Metabolomics for cells typically requires millions of cells, and measures the average metabolic level of the entire tissue. Thus, the current understanding of the heterogeneity of intercellular metabolism is very limited. Although studies have achieved analysis of single cell metabolome, most previous studies have had low throughput and often used very large cells, such as Xenopus eggs or sea rabbit neuronal cells. Mass spectrometry imaging techniques enable large-scale metabolomics of tissues or cultured cells with single-cell spatial resolution, however mass spectrometry imaging has difficulty in distinguishing between intracellular and extracellular metabolites, or attributing metabolic features to specific cell types. Furthermore, none of the previous single cell techniques can analyze rare cell populations, such as stem cells, without a sample enrichment step (e.g., flow cytometry isolation). Therefore, the construction of new tools for metabolic fingerprinting of single-cell samples has an urgent need for the analysis and application of cell heterogeneity of rare cell populations.
Compared with the traditional detection technology, the mass spectrometry detection has high flux and high sensitivity, and can carry out molecular identification and structural analysis. Mass spectrometry is a preferred means of detection and analysis due to its superior properties.
The most common include gas chromatography-mass spectrometer, liquid chromatography-mass spectrometer, and matrix-assisted laser desorption time-of-flight mass spectrometer (MALDI). Because the pretreatment steps are complicated and the time consumption is long, the gas chromatography-mass spectrometer and the liquid chromatography-mass spectrometer are difficult to realize low cost of the single cell sample, and the analysis and the detection of the large sample are applied to the reality. Compared with the two mass spectrum modes, the matrix-assisted laser desorption time-of-flight mass spectrometer has the characteristics of simple sample preparation and high analysis efficiency.
Therefore, those skilled in the art are devoted to develop a single-cell sample fingerprinting molecular detection application technology based on inorganic nanoparticle matrix-assisted laser desorption ionization mass spectrometry.
Disclosure of Invention
In view of the above defects of the prior art, the technical problem to be solved by the present invention is how to develop a novel matrix material and apply the novel matrix material to the detection of the intracellular metabolite fingerprint of the single-cell sample by the matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
In order to achieve the purpose, the invention provides a single cell sample fingerprint spectrum detection method based on inorganic nanoparticles, which is characterized by comprising the following steps:
step 1: preparing an iron oxide micro-nano particle matrix;
step 2: spraying the ferric oxide micro-nano particle matrix into a single-cell sample containing small biological molecules to prepare a sample to be analyzed;
and step 3: performing MALDI mass spectrometry detection on the fingerprint of the sample to be analyzed;
and 4, step 4: analyzing the detection result of the MALDI mass spectrum, and drawing a conclusion.
As one embodiment of the present invention, in step 1, the preparation method of the iron oxide micro-nano particle matrix comprises the following steps:
step 1.1: sequentially adding sodium citrate, ferric chloride and sodium acetate into a solution of ethylene glycol for ultrasonic dispersion, transferring the mixed solution into a Teflon high-pressure reaction kettle, reacting for 8 hours at 100-300 ℃, washing a product with ethanol and deionized water, and finally drying at 60 ℃ for later use to obtain the iron oxide micro-nano particles;
step 1.2: and (3) dispersing the iron oxide micro-nano particles obtained in the step 1.1 in deionized water, and using the iron oxide micro-nano particles as a matrix.
Further, the iron oxide micro-nano particles are dispersed in deionized water to form a solution with the concentration of 1 mg/ml.
In step 1 of the invention, the iron oxide micro-nano particles are spherical and have a diameter of 200-300 nanometers.
In step 2 of the present invention, the molecular weight of the small biomolecule is less than 500Da.
Preferably, the biological small molecule comprises a saccharide and an amino acid.
As one embodiment of the present invention, in step 2, the single cell sample is obtained by performing single cell sorting by a flow cytometer and then performing low temperature lysis.
Specifically, the preparation method of the single cell sample comprises the following steps:
step 2.1: sorting single cell samples of the antibody-labeled target type using a flow cytometer and preparing the single cell samples in 384-well plates containing 80% methanol;
step 2.2: the single cell samples obtained in step 2.1 were lysed in liquid nitrogen for 10 minutes and stored at-80 ℃ for future use.
In the step 3 of the invention, the MALDI mass spectrometry detection adopts a reflection mode and positive ion detection.
The invention also aims to provide the application of the single-cell metabolic fingerprint detection method based on the inorganic nanoparticles in the detection and analysis of the small biological molecules in the single cell.
Compared with the prior art, the invention has the following beneficial effects:
the iron oxide micro-nano particle matrix synthesized by the invention has low preparation cost and simple synthesis steps, and the problems of the traditional organic matrix, such as background interference and hot spot effect of small molecular segments, can be solved by taking the micro-nano particles as the matrix material in the mass spectrum. In addition, in the invention, the single cell sample is obtained by pretreatment steps of single cell sorting by a flow cytometer, low-temperature cracking and the like, and automatic spotting is carried out by utilizing a high-flux pipetting device, so that small molecular substances in the single cell lysate can be efficiently and quickly detected and analyzed.
The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.
Drawings
FIG. 1 is a mass spectrum of the present invention used for matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection of a small molecular weight end of a single cell;
FIG. 2 is a SEM representation picture of the iron oxide micro-nano particles prepared in a preferred embodiment of the invention;
FIG. 3 is a schematic diagram of the present invention for identifying small molecules in single cell samples of different cell types of hematopoietic stem cell lineage by matrix-assisted laser desorption ionization time-of-flight mass spectrometry;
FIG. 4 is a schematic diagram of the present invention for detecting differentially expressed metabolites in single cell samples of different cell types of hematopoietic stem cell lineage by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Detailed Description
The technical contents of the preferred embodiments of the present invention will be more clearly and easily understood by referring to the drawings attached to the specification. The present invention may be embodied in many different forms of embodiments and the scope of the invention is not limited to the embodiments set forth herein.
The first embodiment is as follows: preparation of iron oxide micro-nano particle matrix
The preparation method of the iron oxide micro-nano particle matrix comprises the following steps:
step 1.1: sequentially adding 0.72g of trisodium citrate, 3.0g of ferric trichloride hexahydrate and 4.8g of sodium acetate into a 100mL ethylene glycol solution for ultrasonic dispersion, then transferring the mixed solution into a 200mL Teflon high-pressure reaction kettle, reacting for 8 hours at 200 ℃, sequentially washing the product with 50mL of ethanol and deionized water for 3 times, and finally drying at 60 ℃ for later use to obtain the iron oxide micro-nano particles;
step 1.2: dispersing 1mg of the iron oxide micro-nano particles obtained in the step 1.1 in 1ml of deionized water to form a solution with the concentration of 1mg/ml, and using the solution as a matrix.
Scanning electron microscope results of the prepared iron oxide micro-nano particle matrix are obtained by a NERCN-TC-006 field emission scanning electron microscope, and are shown in figure 2.
As can be seen from FIG. 2, the diameter of the prepared iron oxide micro-nano particles is a spherical material concentrated at 230-270 nm, and the scanning electron microscope result of FIG. 2 shows that the size of the synthesized iron oxide micro-nano particle matrix is uniform and the surface is rough.
The iron oxide micro-nano particles are used as matrix materials in mass spectrum, so that the problems of the traditional organic matrix, such as background interference and hot spot effect of small molecular segments, can be solved.
Example two: preparation of Single cell samples
The preparation method of the single cell sample comprises the following steps:
step 2.1: sorting single cell samples of the antibody-labeled target type using a flow cytometer and preparing the single cell samples in 384-well plates containing 80% methanol;
step 2.2: the single cell samples obtained in step 2.1 were lysed in liquid nitrogen for 10 minutes and stored at-80 ℃ for future use.
Example three: single cell sample fingerprint detection based on inorganic nanoparticles
(1) Spraying the ferric oxide micro-nano particle matrix prepared in the example 1 to the single cell sample containing the small biological molecules prepared in the example 2 to prepare a sample to be analyzed;
(2) Sample preparation, drying and automatic loading are carried out on a mass spectrum target plate by using a 384-hole high-throughput liquid-transferring device; secondly, MALDI mass spectrometry detection is carried out on the fingerprint of the sample to be analyzed, and the model of the instrument is Autoflex-TOF (/ TOF) -MS (Bruker Autoflex Speed); the mass spectrum detection adopts a reflection mode and positive ion detection, and the specific parameters are as follows: the laser wavelength is 355nm, and the laser frequency is 2kHz; the accelerating voltage is 20kV, and the repetition rate of delayed extraction is 1kHz; the delay time is 150ns; each analysis was superimposed with 2000 laser shots.
(3) The MALDI mass spectrometry results were analyzed and concluded to be as shown in fig. 1.
As can be seen from FIG. 1, the invention adopts the single cell sample as the pretreatment steps of single cell sorting by a flow cytometer, low temperature cracking and the like, and utilizes a high-throughput liquid transfer device to carry out automatic spotting, so that the small molecular substances in the single cell lysate can be efficiently and rapidly detected and analyzed.
Example four: fingerprint spectrum mass spectrum detection of hematopoietic stem cell lineage single cell sample
1. Preparing an iron oxide micro-nano particle matrix:
step 1.1: sequentially adding 0.72g of trisodium citrate, 3.0g of ferric trichloride hexahydrate and 4.8g of sodium acetate into a 100mL ethylene glycol solution for ultrasonic dispersion, then transferring the mixed solution into a 200mL Teflon high-pressure reaction kettle, reacting for 8 hours at 200 ℃, sequentially washing the product with 50mL of ethanol and deionized water for 3 times, and finally drying at 60 ℃ for later use to obtain the iron oxide micro-nano particles;
step 1.2: dispersing 1mg of the iron oxide micro-nano particles obtained in the step 1.1 in 1ml of deionized water to form a solution with the concentration of 1mg/ml, and using the solution as a matrix.
2. Preparation of hematopoietic stem cell lineage single cell samples
Step 2.1: sorting the antibody-labeled hematopoietic stem cell lineage single cell samples using a flow cytometer, and preparing the hematopoietic stem cell lineage single cell samples in 384-well plates containing 80% methanol; the schematic diagram of hematopoietic stem cell lineage cell type identification is shown in fig. 3, and the schematic diagram includes 12 kinds of biological small molecules, mainly polysaccharides and amino acids.
Step 2.2: the hematopoietic stem cell lineage single cell sample obtained in step 2.1 was lysed in liquid nitrogen for 10 minutes and stored at-80 ℃ until use.
3. Spraying an iron oxide micro-nano particle matrix into a hematopoietic stem cell lineage single cell sample containing biological micromolecules of 12 polysaccharides and amino acids to prepare a sample to be analyzed;
4. sample preparation, drying and automatic loading are carried out on a mass spectrum target plate by using a 384-hole high-throughput liquid-transferring device; secondly, MALDI mass spectrometry detection is carried out on the fingerprint of the sample to be analyzed, and the model of the instrument is Autoflex-TOF (/ TOF) -MS (Bruker Autoflex Speed); the mass spectrometry adopts a reflection mode and positive ion detection, and the specific parameters are as follows: the laser wavelength is 355nm, and the laser frequency is 2kHz; the repetition rate of the delayed extraction was 1kHz and the acceleration voltage was 20kV; the delay time is 150ns; each analysis was superimposed with 2000 laser shots.
5. The mass spectrum data is preprocessed and subjected to multi-factor analysis, and a single-cell small molecule metabolite with large expression difference is screened out, and the result is shown in fig. 4.
As can be seen from fig. 4, the fingerprint analysis tool for constructing the biomolecular minor molecules metabolized by the single cell sample of the stem cell lineage has a good effect on the analysis and application of the cell heterogeneity of rare cell populations, such as hematopoietic stem cells, can reveal the cell heterogeneity of rare cell populations, such as hematopoietic stem cells, and has great application potential in deep interpretation of the single cell level in the processes of growth and development of organisms, pathogenic mechanisms and the like.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. A unicellular sample fingerprint detection method based on inorganic nanoparticles is characterized by comprising the following steps:
step 1: preparing an iron oxide micro-nano particle matrix;
step 2: spraying the ferric oxide micro-nano particle matrix into a single cell sample containing biological micromolecules to prepare a sample to be analyzed;
and step 3: performing MALDI mass spectrometry detection on the fingerprint of the sample to be analyzed;
and 4, step 4: and analyzing the detection result of the MALDI mass spectrum to obtain a conclusion.
2. The inorganic nanoparticle-based single cell metabolic fingerprint detection method according to claim 1, wherein in the step 1, the preparation method of the iron oxide micro-nano particle matrix comprises the following steps:
step 1.1: sequentially adding sodium citrate, ferric chloride and sodium acetate into a solution of ethylene glycol for ultrasonic dispersion, transferring the mixed solution into a Teflon high-pressure reaction kettle, reacting for 8 hours at 100-300 ℃, washing a product with ethanol and deionized water, and finally drying at 60 ℃ for later use to obtain the ferric oxide micro-nano particles;
step 1.2: and (3) dispersing the iron oxide micro-nano particles obtained in the step 1.1 in deionized water, and using the iron oxide micro-nano particles as a matrix.
3. The inorganic nanoparticle-based single cell metabolic fingerprint detection method of claim 2, wherein: and dispersing the iron oxide micro-nano particles in deionized water to form a solution with the concentration of 1 mg/ml.
4. The inorganic nanoparticle-based single-cell metabolic fingerprint detection method according to claim 1, wherein in step 1, the iron oxide micro-nano particles are spherical and have a diameter of 200-300 nm.
5. The inorganic nanoparticle-based single-cell metabolic fingerprint detection method according to claim 1, wherein in step 2, the molecular weight of the biomolecular is less than 500Da.
6. The method for detecting the metabolic fingerprint of a single cell based on inorganic nanoparticles as claimed in claim 3, wherein in step 2, the small biological molecules comprise saccharides and amino acids.
7. The inorganic nanoparticle-based single cell metabolic fingerprint detection method according to claim 1, wherein in step 2, the single cell sample is obtained by low temperature lysis after single cell sorting by a flow cytometer.
8. The inorganic nanoparticle-based single cell metabolic fingerprint detection method according to claim 7, wherein the single cell sample preparation method comprises the following steps:
step 2.1: sorting single cell samples of the antibody-labeled target type using a flow cytometer and preparing the single cell samples in 384-well plates containing 80% methanol;
step 2.2: the single cell samples obtained in step 2.1 were lysed in liquid nitrogen for 10 minutes and stored at-80 ℃ until use.
9. The inorganic nanoparticle-based single cell metabolic fingerprint detection method of claim 1, wherein in the step 3, MALDI mass spectrometry detection adopts a reflection mode and positive ion detection.
10. Use of the inorganic nanoparticle-based single cell metabolic fingerprint detection method of any one of claims 1-9 for the detection and analysis of small biological molecules in single cells.
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