CN108387634A - Mass spectrum substrate and preparation method and purposes - Google Patents
Mass spectrum substrate and preparation method and purposes Download PDFInfo
- Publication number
- CN108387634A CN108387634A CN201810053331.XA CN201810053331A CN108387634A CN 108387634 A CN108387634 A CN 108387634A CN 201810053331 A CN201810053331 A CN 201810053331A CN 108387634 A CN108387634 A CN 108387634A
- Authority
- CN
- China
- Prior art keywords
- substrate
- sample
- hydrophobic
- hydrophilic
- maldi
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/64—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode using wave or particle radiation to ionise a gas, e.g. in an ionisation chamber
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81B—MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES
- B81B1/00—Devices without movable or flexible elements, e.g. microcapillary devices
- B81B1/006—Microdevices formed as a single homogeneous piece, i.e. wherein the mechanical function is obtained by the use of the device, e.g. cutters
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81B—MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES
- B81B7/00—Microstructural systems; Auxiliary parts of microstructural devices or systems
- B81B7/04—Networks or arrays of similar microstructural devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00015—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems
- B81C1/00023—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems without movable or flexible elements
- B81C1/00119—Arrangement of basic structures like cavities or channels, e.g. suitable for microfluidic systems
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00015—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems
- B81C1/00206—Processes for functionalising a surface, e.g. provide the surface with specific mechanical, chemical or biological properties
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00436—Shaping materials, i.e. techniques for structuring the substrate or the layers on the substrate
- B81C1/00523—Etching material
- B81C1/00539—Wet etching
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
- G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
Abstract
The present invention provides a kind of in MALDI TOF Mass Spectrometer Methods carries the substrate speciallies of BIOMARK biomarkers, and structure includes:Hydrophobic region outside the hole made of chemical surface modification;Hydrophilic region in the circle modified by Specialty Chemical reagent;Titrate the surface circular hole or square hole of BIOMARK samples;The good base material of strong but pliable in texture flatness;The substrate surface of single-sided polishing mirrored effect.In addition, the substrate may also include the Quick Response Code for identification.Micro nano structure substrate provided by the invention can effectively be enriched with sample, and the mass spectrometry profile sample peak accuracy of the uniform growth of crystal orientation, BIOMARK biological detections is high, and signal-to-noise ratio is high, baseline bottom.
Description
Technical field
The improvement substrate that the present invention relates to a kind of for MALDI-TOF MS and with micro nano structure, belongs to Mass Spectrometer Method
Field.
Background technology
MALDI-TOF-MS is Matrix Assisted Laser Desorption Ionization Time of
The english abbreviation of Flight Mass Spectrometry, i.e. matrix solid-dispersion flight time mass spectrum, also known as
MALDI TOF, it is the novel organic mass spectrometry of a kind of soft ionization developed in recent years, by introducing substrate molecule, is made to be measured
Molecule does not generate fragment, solves the problems, such as non-volatile and thermal instability large biological molecule desorption ionization, is that analysis is difficult
One of the important means of organic substance of volatilization.The development that MALDI TOF are worldwide advanced by leaps and bounds, and extensively
Bioanalysis and chemical detection and security department applied to biotechnology and the drug development, scientific research field of pharmacy corporation
Nuclear radiation, the monitoring etc. of chemical substance and bio-pathogen.
The principle of MALDI-TOF MS is:When thin with the laser irradiation sample of some strength and substrate formed cocrystallization
Film, matrix absorb energy from laser, sample desorption, and electric charge transfer occurs between matrix-sample and makes ionized sample molecule,
The sample of ionization accelerates to fly over dirft tube under electric field action, is detected according to the flight time difference for reaching detector,
The ratio between the quality charge (M/Z) for measuring ion directly proportional to the flight time of ion detects ion.MALDI-TOF MS's
Center Technology is exactly that the difference of the mass-to-charge ratio (m/z) according to sample is detected, and measures the molecular weight of sample molecule.According to
The principle of MALDI-TOF MS, MALDI-TOF MS have high sensitivity, accuracy height, high resolution, collection of illustrative plates simplicity, quality model
Enclose the features such as wide and speed is fast, operationally sample preparation it is easy, can milligram ammonia, extensive, parallelization and increasingly automated processing wait for
Biological sample is examined, and has special superiority measuring large biological molecule and synthetic high polymer application aspect.It has become in recent years
For the strong work of detection and identification polypeptide, protein, polysaccharide, nucleotide, glycoprotein, high polymer and a variety of synthetic polymers
Tool.After such as applying MALDI-TOF MS to measure the peptide mass fingerprinting spectrum (PMF) of protein digestion, source cracking (PSD) fragment from
Mass spectrum web database search is composed and combined to subgraph, can get the sequence of polypeptide, protein.Using MALDI-TOF to genome
Single nucleotide polymorphism (SNPs) carries out analysis detection, can distinguish and differentiate relative molecular mass up to 7,000 or so (containing more than 20
Base), there is only the different DNA of 1 base difference.It is worth pointing out that MALDI-TOF has become life science
One of indispensable important key technology in proteome research.
MALDI-TOF can solve in current protein science following several potential challenges, i.e., the discovery of biomarker,
Molecule diagnosis research and development, protein high throughput analysis and proteomic map, but it is not limited to this, MALDI-TOF MS are applied to
The discovery of biomarker and pathogenic mechanism is disclosed on molecular level, to be applied to molecule diagnosis, target is controlled
It treats and personalized medicine, growth, the rule of development and the vital movements such as metabolic regulation and tight will be more completely prompted for people
The genesis mechanism of weight disease carries out the diagnosis prevention of disease for the mankind and new drug development provides important theoretical foundation.
MALDI-TOF, can due to having the features such as quickly analysis, operation simple, automation, amount of samples is few and high-throughput
Directly detect serum, blood plasma, urine, cerebrospinal fluid, articular cavity synovia, bronchus eluent, cell pyrolysis liquid, tissue extract and
Various secretion form the major disease, the proteomics of important living resources, protein of scale using MALDI-TOF
Group fingerprint or mass spectrum peptide figure investigative technique platform, can establish the proteome databases with independent intellectual property rights, find
The GAP-associated protein GAP of disease is composed and the biomarker spectrum with important application foreground, to disclose the mechanism of disease, as
Early diagnosis, molecule parting, curative effect and Index for diagnosis foundation, and find out the molecular target for being likely to become new drug design, be disease
Disease provides new therapeutic scheme.The analyses such as proteomic map, mass spectrum peptide figure, biomarker spectrum based on MALDI-TOF are examined
Disconnected development by be molecular medicine field a revolution, it provides not only a kind of new diagnostic method, and with very high
Feasibility, it can measure a results up to ten thousand from micro blood within several minutes of times and analyze it.
In this regard, the present inventor uses first, MALDI-TOF MS carry out the separation of the biological samples such as serum, polypeptide spectrum is quickly swept
Retouch and be sequenced with genius morbi peak, in the important physiology of the research mankind and pathologic process important biomolecule marker spectrum experiment confirmation and point
Analysis technology obtains several with the relevant important biomolecule marker spectrum of mankind's major disease, the early warning applied to major disease
Judge with the course of disease.As the inventors discovered that for finding that malignant tumour blood serum designated object is composed, creating has independent intellectual property right
China common cancer crowd " serum mass spectrum polypeptide spectrum ", obtain tumour early warning, the mass spectrometric data that the course of disease judges
Standard, for example, Chinese granted patent 200810172142.0, " a kind of preparation method for detecting liver cancer characteristic protein model ",
Chinese granted patent 200810147419.4, " preparation method for detecting brain glioma characteristic mass spectra model ", China
Granted patent 201110216986.2, " mass spectra model and construction method for detecting lung cancer albumen ".In addition, according to mass spectrum skill
Art is for nucleic acid and determinand (such as microorganism, particular source ingredient) or symptom (saprodontia, senile dementia, hereditary hearing impairment)
In research, the present inventor also obtains numerous Chinese granted patents, such as Chinese granted patent 201310158363.3, " preparation
The method of bacteria nucleic acid fingerprint characteristic spectrum library ", Chinese granted patent 201310368422.X, " the mass spectrum mould of detection saprodontia albumen
Type and construction method ", Chinese granted patent 201310175514.6 " are used to detect and the relevant gene SNP of hereditary hearing impairment
Primer system and application thereof ".Therefore, the disease related data for MALDI-TOF MS development for " serum mass spectrum polypeptide spectrum "
The bioinformatics tools and medical diagnosis on disease software of excavation, can be applied to clinic and introduce to the market.No matter for scientific research field, doctor
Treatment system, or biological market all have broad prospects.
Due to forming ultra-thin and uniform sample crystallization on the detection carrier of MALDI-TOF (target plate or substrate), always
It is to improve one of the research direction of mass spectrum plot quality, therefore some relevant researchs occur.Chinese granted patent
201410090967.3, " the alternate micro-array chip of hydrophobe and its method for mass spectrum imaging quantitative analysis is prepared " to provide
It is a kind of to prepare detection chip by the way that hydrophobic and hydrophilic region is arranged on chip, wherein will be hydrophobic using screen printing technique
Property polymer (dimethyl silicone polymer or polymethyl methacrylate) according to designed template brush on electro-conductive glass, wherein
Applying area is hydrophobic region, and clear area is as reserved hydrophilic area.Then, it is coated with water wetted material (such as aqueous solution of sample system)
Clear area forms hydrophilic area and the hydrophobic region at uniform interval.This method carrys out design template shape by using screen printing technique,
It leaves some space area (also known as " being left white processing ") in hydrophilic area in advance, it is therefore desirable to special printing equipment and operating software, simultaneously
When being coated with hydrophobic region, target plate will absolutely be stood.Meanwhile the precision of screen printing technique is low, the boundary of close and distant water area cannot reach
To micron accuracy.Furthermore, it is desirable to which testing mixture system aqueous solution is equably layered on hydrophilic area, chip baking and curing is placed in
60 degrees Celsius of solidification 2h, increase production time and cost in baking oven.
Chinese granted patent 201110401165.6, " method for carrying out enrichment and desalination purification processing to biological sample " are public
A kind of method preparing the detection target plate with closed pattern surface using hydrophobic and water wetted material is opened, including by base
It is constructed on bottom and is used as barrier with larger-diameter polymer coating (such as polymethyl methacrylate, polystyrene or photoresist)
Circle area, is then attached to as hydrophobic layer in entire substrate using fluorine-containing monolayer or evaporated metal layer.Since the hydrophobic layer cannot
It is adhered to above-mentioned barrier circle area, therefore target plate is immersed in organic solvent and is ultrasonically treated, barrier circle area can be removed.Most
Afterwards, then by the polymer coating round area's internal modifications small diameter concentric circles, it is (fluorine-containing to obtain hydrophobic outer ring
Monolayer or evaporated metal layer)-hydrophilic hydrophobic the inner ring of centre circle-(polymer coating) concentric circles mass spectrum target plate, because
This this method is also known as " barrier coating ".However, this method constructs the closed pattern of hydrophobe-hydrophile-hydrophobic region in substrate
Surface needs to carry out two step hydrophobic treatments, that is, uses fluorine-containing reagent 1~5 hour of heat growth, work at 100~250 DEG C
Skill is complicated, and process takes.Since this method needs to accurately control the spacing of polymer coating twice, spacing is too small cause outer ring and
Inner ring is connected, simultaneously because being mutated different substrate surfaces respectively using two different hydrophobic materials, what is obtained is hydrophobic
The contact angle of area's drop is too small, causes hydrophobic effect poor, affects the application of common lab.
Chinese patent application 200610023671.5 discloses " a kind of low-abundance protein target previous step desalination and enrichment
Method ", wherein by hydrophobic polymer (polymethyl methacrylate, polyethylene, polystyrene, polyvinyl fluoride etc.) in advance in target
The sample cell center portion of plate is coated, and then carries out albumen point sample so that protein sample is enriched on hydrophobic polymer layer.
Then, point sample area is added in matrix solution so that pollutant and inorganic salts in protein sample are spread out, and finally obtain sample
With the crystallization of matrix uniform and delicate.Since this method is that protein sample is directly added to hydrophobic layer, by the way that excessive matrix is added
Solution is purified, although objectively there is certain purification effect, causes the waste of protein sample.Meanwhile if manually
Operation in the aperture of each Kapton films, takes 0.2 μ l hydrophobic polymers solution points, and the precision of 0.2 μ l solution is difficult
With control, the hydrophobic homogeneity of orifice surface is influenced;If be automatically brought into operation, corresponding point sample equipment is needed to configure, increases cost, is prepared
Complexity is not appropriate for the detection and application of the trace protein sample for being not easy to prepare.
Due to above-mentioned target plate it is first research in or target plate surface there is no hydrophilic-hydrophobic difference, (such as Chinese patent
200610023671.5), cause crystal habit poor, or form the coating of hydrophilic-hydrophobic difference, but hydrophobe boundary cannot reach
It is too small to micron accuracy or liquid-drop contact angle, but preparation process is excessively complicated, and cannot save time and cost (such as Chinese patent
201410090967.3, patent 201110401165.6), or additional device and inspection software are needed, or need excessive
Precious albumen sample be detected, the accuracy that these result in Mass Spectrometer Method sample peak is low, and signal-to-noise ratio is low, and baseline is high.
In addition, with MALDI TOF the discovery of biomarker, molecule diagnose research and development, protein high throughput analysis and
The extensive use in proteomic map field provides relatively simple, convenient and cheap target plate and also becomes current demand.For
This provides one kind in the present inventor's prior authorization patent 201520142252.8, " hydrophobicity of Mass Spectrometer Method concentrates target plate "
Prepare the simple and easy method of target plate, wherein using the target plate main body of stainless steel or aluminium alloy, by using hydrophobic material (as gathered
Propylene or polyethylene contain based polymers) pad pasting or coating target plate surface.Wherein, which is equipped with and sample room bottom
The shape or structure of logic relationship, and by the combination of different horizontally-arranged and perpendicular row marks, to position different samples.Due to containing
Have the matrix of sample in spotted area because hydrophobic effect is assembled, can reduce amount of samples, and by after drying crystalline again
Mass spectrum is carried out, good effect is obtained.It is demonstrated experimentally that the hydrophobicity with above-mentioned specific shape and structure concentrates target plate, hence it is evident that
Better than single metallic plates, while having the advantages that Sample location is accurate, meets mass spectrum requirement.
As noted previously, as the sample on target plate, which forms well-defined crystal, can be conducive to obtain on certain the mass spectrum of high quality
Detection figure, and the considerations of for production cost, obtain can high quality crystallization and prepare simply and cheap mass spectrum target plate at
The research purpose to deepen continuously for technical staff.
Invention content
In order to overcome traditional Mass Spectrometer Method target plate surface there is no hydrophilic-hydrophobic difference, cause crystal habit poor, Mass Spectrometer Method
The accuracy at sample peak is low, and signal-to-noise ratio is low, and baseline is high, while the disadvantage that manufacturing cost is excessively high and complicated, liquid-drop contact angle are too small
The defect of hydrophobic performance is influenced, the present invention provides a kind of improvement substrate detecting and have micro nano structure for MALDI-TOF,
By increasing the hydrophilic and hydrophobic difference of target plate on surface, so that sample to be tested is formed well-defined crystal in substrate location hole, carry
High Mass Spectrometer Method spectrogram quality and qualification result accuracy.
Therefore, an object of the present disclosure is to provide a kind of improvement detecting and have micro nano structure for MALDI-TOF
Substrate, include with the hydrophobic region substrate outside hydrophilic point sample area, point sample area, wherein:
The surface of the substrate is handled by silane coupling agent, to form hydrophobic region;
The point sample area is handled by acid reagent, silane coupling agent is removed, after concurrent biochemical reaction modification, to be formed
Hydrophilic area.
In another embodiment, the substrate is stainless steel strong but pliable in texture, that flatness is good, diamond, monocrystalline silicon, stone
The substrate of English crystal.In a specific embodiment, the substrate first passes through chemically mechanical polishing (chemical in advance
Mechanism polish, CMP) it is handled with single-sided polishing mirrored effect.By CMP method, substrate is thrown twice
Light, rough polishing and fine polishing.The purpose of rough polishing is the removal remaining mechanical damage of substrate surface, is generally removed within the scope of 30um from surface
Thickness.The purpose of fine polishing is that removal polishes the slight damage left in substrate surface and cloud defect for the first time, generally from surface
Remove 2~3um.There is mirrored effect by the substrate surface polished twice, during Mass Spectrometer Method, pass through illumination, light path
Reflection and camera real-time display, can directly observe the case where substrate surface sample is bombarded, change the position of laser bombardment,
Obtain best collection of illustrative plates.In a more specific embodiment, the substrate is the silicon chip or quartz substrate of single-sided polishing.
In one embodiment, the silane coupling agent is selected from vinyl silanes, amino silane, dimethyl dichloro silicon
Alkane.Wherein, dimethyldichlorosilane growth hydrophobic membrane is easy to operate, and contact is larger, and hydrophobicity is good, and hydrophobic layer thickness, which can reach, to be received
Rice is to micron dimension, therefore silane coupling agent is preferably dimethyldichlorosilane.In a specific embodiment, by the idol
Join the hydrophobic surface of agent entirety silanization, 120 ° of contact angle > or 130 ° of > or 140 ° of >, in a more preferred embodiment,
150 ° of the contact angle >, to form super hydrophobic surface.In other preferred embodiments, the thickness of the hydrophobic surface exists
Nanometer arrives micron dimension.
In other embodiments, the point sample area, which is drawn, is divided into the rectangular of the suitable sample rooms MALDI-TOF specification
Shape draws and is less than 10 μm every precision.In a specific embodiment, lithography mask version is made, hydrophobe is formed in substrate surface
Region (see Fig. 1), boundary precision reach micron level.The acid reagent is selected from hydrofluoric acid (HF), sulfuric acid (H4SO2), hydrogen bromide
(HBr), preferably hydrofluoric acid (HF).In another embodiment, by the acid reagent by preparing, hole inner region is dropped in
Domain, after chemical reaction modification so that hydrophily is presented in hole inner region.
In any of the above-described embodiment, the substrate has the two-dimension code area for substrate identification, wherein logical
Photoetching or etching are crossed to print two-dimension code pattern, is at least divided into 8 × 8~15 × 15 array.In a specific embodiment,
Laser beam is controlled by computer to arrive in nanometer in etching substrate or the preset two-dimension code pattern of photoetching, the thickness profile
Micron dimension.In another embodiment, it is described be used for substrate identification, refer to Quick Response Code and patient biological information or
The source-information of sample is associated with.In other embodiments, while carrying out Mass Spectrometer Method, code reader or code reader can be used to read
Quick Response Code, and the source-information and corresponding mass spectrometric data of the corresponding patient's biological information of substrate or sample are inputted into electricity simultaneously
Brain is encoded, stored and is identified.
In one embodiment, the hydrophilic point sample area quantity on the substrate is 48,56,70,96,384 or other numbers
Amount, hydrophilic area shape can select circle, square, triangle, polygon etc..In a specific embodiment, the parent
Water area size range is 100 × 100 μm to 1 × 1mm, and the sizes of substrate is 20 × 30-83 × 125mm.
In above-mentioned all embodiments, wherein the substrate is the substrate for MALDI-TOF BIOMARK detections.
Second purpose of the invention is to provide the preparation method for the MALDI-TOF micro nano structure substrates detected, including:
(1) select the substrate strong but pliable in texture, flatness is good as substrate;
(2) substrate is handled with silane coupling agent
(i) substrate is cleaned:Priority is immersed into acetone, methanol aqueous solution, chloroform, after being respectively washed, takes out drying;
(ii) in a heated condition, concentrated acid and hydrogen peroxide are slowly added in the aqueous solution containing substrate, carry out abundant oxygen
After changing reaction, cleaned in chloroform, ultra-pure water respectively, which can be repeated several times;
(iii) it places the substrate in clean container, the silane coupler solution hydrolyzed in advance is added, and ammonium hydroxide is added and urges
Change, until Silanization reaction is fully completed;
(iv) substrate is taken out, is respectively placed in ethyl alcohol, ultra-pure water, chloroform, is cleaned by ultrasonic;
(v) by the substrate after cleaning, the substrate of 120 ° of contact angle > is measured and selects, as qualified substrate;
(3) the hydrophily processing in point sample area:Modulated acid reagent is directly dropped in point sample area, is tried by the acidity
After agent is chemically modified surface, hydrophily cavernous structure is formed, is then respectively placed in ethyl alcohol, ultra-pure water, chloroform, ultrasound
Cleaning.
In one embodiment, step (1) described substrate is stainless steel strong but pliable in texture, that flatness is good, diamond, list
The substrate of crystal silicon, quartz crystal.In a specific embodiment, the substrate first passes through chemically mechanical polishing in advance
(chemical mechanism polish, CMP) has single-sided polishing mirrored effect by processing.By CMP method, to substrate
It is polished twice, rough polishing and fine polishing.The purpose of rough polishing is the removal remaining mechanical damage of substrate surface, is generally removed from surface
Thickness within the scope of 30um.The purpose of fine polishing is that removal polishes the slight damage left in substrate surface for the first time and cloud lacks
It falls into, generally removes 2~3um from surface.There is mirrored effect by the substrate surface polished twice, during Mass Spectrometer Method,
By illumination, light path reflection and camera real-time display, the case where substrate surface sample is bombarded can be directly observed, changes and swashs
The position of light bombardment, obtains best collection of illustrative plates.In a more specific embodiment, the substrate be single-sided polishing silicon chip or
Quartz substrate.
In another embodiment, step (2) described silane coupling agent is selected from vinyl silanes, amino silane, diformazan
Base dichlorosilane.Wherein, dimethyldichlorosilane growth hydrophobic membrane is easy to operate, and contact is larger, and hydrophobicity is good, hydrophobic layer thickness
Nanometer can be reached to micron dimension, therefore silane coupling agent is preferably dimethyldichlorosilane.In a specific embodiment,
By the hydrophobic surface of the coupling agent entirety silanization, 120 ° of contact angle > is in a preferred embodiment, described to connect
150 ° of feeler >, to form super hydrophobic surface.In other preferred embodiments, the thickness of the hydrophobic surface is arrived in nanometer
Micron dimension.
In other embodiments, step (3) the point sample area, which is drawn, is divided into the suitable sample rooms MALDI-TOF specification
Rectangle, draw every precision be less than 10 μm.In a specific embodiment, lithography mask version is made, is formed in substrate surface
Close and distant water area (see Fig. 1), boundary precision reach micron level.The acid reagent is selected from hydrofluoric acid (HF), sulfuric acid (H4SO2)、
Hydrogen bromide (HBr), preferably hydrofluoric acid (HF).In another embodiment, by the acid reagent by preparing, hole is dropped in
Inner region, after chemical reaction modification so that hydrophily is presented in hole inner region.
In any of the above-described embodiment, step (4) described substrate has the two-dimension code area for substrate identification,
Two-dimension code pattern is wherein printed by photoetching or etching, is at least divided into 8 × 8~15 × 15 array.It is embodied at one
In scheme, laser beam is controlled in etching substrate or the preset two-dimension code pattern of photoetching, the thickness profile by computer
In nanometer to micron dimension.In another embodiment, described to be used for substrate identification, refer to Quick Response Code and the life of patient
The source-information of object information or sample is associated with.In other embodiments, while carrying out Mass Spectrometer Method, with code reader or it can sweep
Code device reads Quick Response Code, and the source-information and corresponding mass spectrometric data of the corresponding patient's biological information of substrate or sample is same
When input computer encoded, stored and identified.
In one embodiment, the hydrophilic point sample area quantity on the substrate is 48,56,70,96,384 or other numbers
Amount, hydrophilic area shape can select circle, square, triangle, polygon etc..In a specific embodiment, the parent
Water area size range is 100 × 100 μm to 1 × 1mm, and the sizes of substrate is 20 × 30-83 × 125mm.
Third purpose of the present invention is to provide the substrate prepared by the above method.
In one embodiment, the substrate is the substrate for MALDI-TOF BIOMARK detections.
4th purpose of the invention is to provide the substrate for Mass Spectrometer Method biomolecule or biomarker sample
Purposes.
In one embodiment, the substrate is the substrate for MALDI-TOF BIOMARK detections.In another implementation
In scheme, the biomolecule biomarker is albumen or polypeptide sample, nucleic acid samples.In one embodiment,
In when carrying out Mass Spectrometer Method, code reader or code reader can be used to read Quick Response Code, and by the corresponding patient's biological information of substrate or
The source-information of sample and corresponding mass spectrometric data input computer simultaneously and are encoded, stored and identified.
Term and definition
Hydrophobicity (hydrophobicity):In chemistry subject, hydrophobicity refers to a molecule (hydrophobe) and water
Mutually exclusive physical property.Hydrophobic molecule is partial to nonpolarity, and therefore compared with can be dissolved in neutral and non-polar solution (such as
Organic solvent).Hydrophobic molecule would generally bunch up in water, and water can then form one at the surface of hydroholic solution
Prodigious contact angle forms drops.For example, hydrophobic molecule includes alkane, oil, fat and majority containing grease
Substance.
Hydrophily (hydrophilic property):The molecule for referring to molecular band polarized group has water big affine
Ability can attract hydrone, or be dissolved in water.The surface for the solid material that this kind of molecule is formed, is easily soaked by water.Have
This characteristic is all the hydrophily of substance.
Contact angle (contact angle):Refer to that the tangent line of the liquid-vapor interface made by gas, liquid, solid three-phase point of intersection is worn
The angle theta between liquid and solid-liquid boundary line is crossed, is the measurement of wetness degree.If θ<90 °, then the surface of solids is hydrophilic,
I.e. liquid is easier to wetting solid, and angle is smaller, indicates that wetability is better;If θ>90 °, then the surface of solids is hydrophobic, i.e. liquid
Body is not easily wetted by solid, is easy to move on the surface.There are many assay method of contact angle, angled mensuration (drop angle
Mensuration), length/height mensuration, power mensuration etc..Wherein drop angular measurement is most common, i.e., in flat surface
A droplet is dripped, it can measurement angle size using the protractor of low-powered microscope.
BIOMARK, i.e. biomarker, microbial identification, protein fingerprint spectrum detection, Genotyping and gene mutation are examined
It surveys, is the BIOMARK biomarkers for finding variation after all, with target plate or substrate bearing, be sent into MALDI-TO F samples
Room, is detected analysis, and structure and the index of substrate directly affect the quality of testing result.The MALDI-TOF BIOMARK,
Can also be the associated mass spectrometry instrument researched and developed based on this or matter either detecting the technology of BIOMARK using MALDI-TOF
Compose the title of product, such as the Related product of Fluidigm companies of U.S. research and development.
Silane coupling agent:Refer to a kind of organic compound with special construction, containing organo-functional group, Inorganic functional groups,
Simultaneously binding force is generated with cathode, apolar substance.The chemical general formula of silane coupling agent is Y-R-SiX3, and Y is to pass through carbon in formula
The non-hydrolyzable organo-functional group that atom is connected with silicon can react to improve compatible with the resin in binder body
Property, such as amino, vinyl, epoxy group, sulfydryl, acryloxypropyl;R is the carbochain with saturation and unsaturated bond, by Y and
Si atoms connect;X is hydrolization group, such as halogen family, alkoxy, isopropyl alkenyloxy group.The silicon that these groups hydrolyze to form
Alcohol can be reacted with the oxide or alkyl of metal or nonmetallic surface, to form Si-O-Si tri- in metal or nonmetallic surface
The silane for tieing up network structure not, prevents metal or nonmetallic corrosion, surface from hydrophobicity is presented.Suitable for the silane coupled of the present invention
Agent is only limited to amino silane (such as aminopropyl triethoxysilane), vinyl silanes (vinyltriethoxysilane) and diformazan
Base dichlorosilane, wherein it is preferred that silane coupling agent is dimethyldichlorosilane.
MALDI-TOF-MS is Matrix Assisted Laser Desorption Ionization Time of
The english abbreviation of Flight Mass Spectrometry, i.e. matrix solid-dispersion flight time mass spectrum, also known as
MALDI TOF, it is the novel organic mass spectrometry of a kind of soft ionization developed in recent years, by introducing substrate molecule, is made to be measured
Molecule does not generate fragment, solves the problems, such as non-volatile and thermal instability large biological molecule desorption ionization, is that analysis is difficult
One of the important means of organic substance of volatilization is widely used for point of the BIOMARK of protein sample and nucleic acid samples now
In son detection.Technology similar with MALDI TOF, Protein-based tumor biomarker flight time mass spectrum (SELDI-TOF-MS)
Flight time mass spectrum is a kind of Identification of Fusion Protein technology and methods that comparison is new.Its most attractive spot is its separation by sample
With mass spectrum (being exactly chromatography) identification albumen combination so that sample direct loading in the state of without crude separation, this makes
Much protein sample (such as serum) can be quickly obtained identification.But the defect of this method is:(1) for different samples
This, take or design several chips according to the target of detection, theoretically can all same nature protein captures, but
It is actually still to have a small amount of molecule not combined with surface-probe.(2) SELDI-TOF-MS is used, is only capable of providing point of protein
Son amount, cannot provide C-terminal, the sequence of N-terminal, cannot also know the configuration of protein, it is therefore desirable to fully after purification by protein,
With the protein on protease digestion chip, peptide fragment is analyzed, then protein sequence is identified with bioinformatics method.(3) party
Method is relatively suitble to clinical diagnosis, but cannot carry out Sequence Identification and the functional study of protein.Therefore, used in this method
BIOMARK substrates cannot be general with MALDI TOF.
The source-information of " Quick Response Code is used for substrate identification ", the biological information or sample that refer to Quick Response Code and patient closes
Connection.For example, the biological information of patient may include but be not limited to, patient's case, living habit, personal background can be specifically diseases
The diagnosis records of people, the familial inheritance relationship of patient, life style of patients data (whether smoke, drink, staying up late, Long-term taking medicine,
Occupy various radiation environments etc.).
The source-information of sample can include but is not limited to, the environmental information of sample or the product information of sample.Such as sample
The environmental information of product can include but is not limited to:The geographical location information of sample, geographic resource information, moves hydrology soil information
The biological informations such as plant and microorganism;Or, including environment germ pollutant, albumen or polypeptide pollutant, disengaging port pollutant
Albumen or polypeptide mass spectrometric data information.
For example, the product information of sample can include but is not limited to:The albumen or polypeptide of food, agricultural product, industrial kind
The mass spectrometric data information of pollutant or impurity.
Technique effect
1, preparation method of the present invention has the characteristics that prepare simple, quick, can manually be prepared, and special dress is not needed
It sets, is suitable for the large-scale use of common lab.
2, present invention introduces Quick Response Codes to realize substrate identification, is associated with the biological information of patient, passes through computer
Real time monitoring and information processing, remote management are suitable for automating high-throughput hospital and use.
3, it is crystallized compared to the sample protein of Conventional substrate, can effectively be enriched with sample, the uniform growth of crystal orientation, BIOMARK
The mass spectrometry profile sample peak accuracy of biological detection is high, and signal-to-noise ratio is high, baseline bottom.
4, the present invention can use the silane coupling agent of technical grade as hydrophobic material, effect with it is other known hydrophobic
Property material quite even more preferably, be not necessarily to hot setting, save time and cost, substrate surface easy to operate and prepared
Water droplet contact angle can reach 130~155 °, have excellent hydrophobic performance.
5, compared to traditional hydrophobic and hydrophilic treatment method such as screen printing technique, the present invention passes through lithography mask version system
Standby hydrophilic-hydrophobic region, boundary precision reach micron dimension, keep sample crystal habit regular, sharpness of border.
6, compared to the three-step approach of traditional hydrophobic and hydrophilic treated substrate " barrier coating ", the present invention actually belongs to
Two-step method, i.e., hydrophobic coating-hydrophilic coating, does not need special device and accurate processing procedure, process is relative to easier.
7, substrate of the invention carries out single-sided polishing processing using CMP (chemical mechanism polish) method,
Make substrate surface that there is mirrored effect, it, can by illumination, light path reflection and camera real-time display during Mass Spectrometer Method
Directly to observe the case where substrate surface sample is bombarded, change the position of laser bombardment, obtains best collection of illustrative plates.
Description of the drawings
Fig. 1, Fig. 2 are a kind of micro nano structure substrate schematic diagram for MALDI-TOF detections, and Fig. 1 is square hole position,
Fig. 2 is round hole position;Including three subregions:1, hydrophilic region;2, hydrophobic region;3, two-dimension code area;
Fig. 3 is that substrate schematic diagram is cleaned in concentrated acid solution.
Fig. 4 is BIOMARK crystal habit contrast schematic diagrams, and it is about 135 ° that wherein lower-left figure, which is substrate J1 surface contact angles,
Sample crystallization figure, bottom-right graph are the sample crystallization figure that substrate J2 surface contact angles are about 155 °.
Fig. 5 is that MALDI-TOF detects the spectrogram comparison of sample peak.
Specific embodiment
The present invention will be described further in conjunction with specific embodiments, these examples are for illustration purposes only, rather than
Limit the scope of the invention.
Embodiment one prepares substrate
(1) substrate material selects
It is characterized in that strong but pliable in texture, flatness is good;Such as stainless steel, diamond, monocrystalline silicon, quartz crystal etc.;Draw be divided into it is suitable
The rectangle of the sample rooms MALDI-TOF specification draws to ensure quality testing precision and is less than 10 μm every precision.
(2) silanization treatment
1, substrate is cleaned
Substrate is placed in a beaker, tiling is opened, and not overlapped, with acetone rinsing substrate, the substrate transfer after cleaning
Into a new beaker.
The mixed liquor of a certain amount of methanol and water is added into beaker, ultrasound, time 30min.
Substrate is transferred in new beaker, a certain amount of chloroform is added, ultrasound, time 30min.
It takes out, dry substrate.
2, substrate is cleaned with concentrated acid solution
As shown in figure 3, placing the substrates in clean beaker, beaker is fixed on holder.
Prepare sink, fill water, is placed below beaker, heating;
By concentrated acid and hydrogen peroxide according to a certain percentage (1:5~1:20) it is slowly added into the beaker equipped with substrate, waits for constantly
There are minute bubbles, oxidation reaction starts, reaction time 40min.Note that needing to be handled with care under conditions of draught cupboard is divulged information.
Pay attention to personal protection.
Substrate is taken out, is placed in the beaker for filling chloroform, ultrasonic 2min;
Substrate is taken out, is placed in the beaker for filling ultra-pure water, ultrasonic 2min.
Repetition step 8,9, until cleaning is complete.
Liquid waste processing:A large amount of water are added in concentrated acid solution, slowly dilutes, NaOH is used in combination to neutralize.
3, silanization treatment
It places the substrates in clean beaker.
Silane coupling agent is hydrolyzed, according to proportioning silane coupling agent:Water:Ethyl alcohol=1:1:8 mixing, wherein silane are even
Join the dimethyldichlorosilane solution that agent is concentration 5%~10%, water uses deionized water, concentration of alcohol 99%.
Silanizing solution after hydrolysis is taken into appropriate be added in the beaker containing substrate;
A concentration of 13%~30% ammonia-catalyzed is added, matches ammonium hydroxide:Silanization=1:5, accelerate reaction to carry out.
The solution that surface is generated to white cigarette is placed in ventilating kitchen, and reaction continues at least 30 minutes time.
4, it is cleaned after silanization
Substrate is taken out, is placed into the new beaker for filling ethyl alcohol, ultrasound, time 10min.
Substrate is taken out, is placed on into the new beaker for filling ultra-pure water water, ultrasound, time 10min.
Substrate is taken out, is placed on into the new beaker for filling chloroform, ultrasound, time 10min.
After dry substrate, 1 μ l water droplets are dripped in substrate surface, it is big to measure water droplet contact angle using the protractor of low-powered microscope
It is small, respectively obtain about 135 ° of contact angle, 155 ° of two kinds of substrate first products.
(3) hydrophilic treated
1, it according to the hole of substrate surface position and size, makes and covers template, metal or glass board material, by hydrophobic region outside hole
It covers, or is directly controlled by computer, carry out hydrophilic reagent point sample moditied processing;
2, Hydrophilic modification:Hydrofluoric acid HF reagents are chosen, after preparation, drop in hole inner region, after chemical reaction modification,
Hydrophily is presented in hole inner region;
3, it cleans
Chemically modified substrate is put into the clean beaker for filling ethyl alcohol, ultrasound, 5min;
Substrate is taken out, is put into the clean beaker for filling chloroform, ultrasound, 5min;
Substrate is taken out, is put into the crystallization beaker for filling pure water or ultra-pure water, ultrasound, 5min;
(4) Quick Response Code marks
The two-dimension code area of square, is at least divided into 8 × 8~15 × 15 array, and minimum cell size micron dimension swashs
Two-dimension code pattern is made in photoetching printing, and thickness is nanometer to micron dimension, and MALDI-TOF Sample Rooms sweep Quick Response Code, identify substrate body
Part;
By above-mentioned steps, about 135 ° of contact angle, 155 ° of two kinds of substrate finished products are obtained.
Embodiment two, the observation of the sample crystallization of target plate and comparison
The matrix solution prepared is taken and drops in about 135 ° of contact angle, 155 ° two kinds of substrates J1, J2 respectively in right amount, and
On two pieces of traditional target plate T1, T2;Matrix naturally dry, then suitable sample drop is drawn on J1, J2, T1, T2, sample dries in the air naturally
After dry, microscopically observation crystal habit is shown in Fig. 4.
Wherein, traditional target plate is the model TO-488 genetic test target plates of SHIMADZU productions.
As shown in figure 4, traditional biological detection target plate T1, T2 crystal habit (see Fig. 4) shape is irregular, lines are not round and smooth,
In coffee ring status, surface is uneven.Due to intermediate empty, surrounding is thick, and when laser bombardment sample, sample peak poor accuracy is made an uproar
Sound pitch, baseline are high.
And regular circle is presented in biological detection substrate J1, J2 crystal habit (see Fig. 4) shape of the present invention, surface is mellow and full
Such as jade, quality rule is uniform, and crystal is respectively careful to growing, and is ideal protein crystal form.Wherein, lower-left figure J1 contact angles
About 135 °, about 155 ° of bottom-right graph J2 contact angles, in comparison, the quality of J2 ratios J1 are more uniform, and crystallization is smooth.
Embodiment three, Mass Spectrometer Method effect compare
It will be covered with the substrate J1 and traditional target plate T1 of gene samples, is put into MALDI-TOF mass spectrographs and detects.
Parameter setting:
Turing mode:linear
Mass Range:3000-9000
Max Laser Rep Rate:10.0
Power:90
Profiles:40
Shots:10
Compare the testing result of two kinds of target plates.
As shown in figure 5, T1 target plate testing results baseline is high, appearance is few, poor accuracy, and signal-to-noise ratio is low;J1 substrates detection knot
Fruit baseline is low, and appearance is more, and accuracy is high, and signal-to-noise ratio is high.
The measurement of example IV, the hydrophobic layer of substrate and hydrophilic layer
The micro nano structure substrate of the present invention, hydrophobic layer cross section measure under Electronic Speculum, 800nm-1 μm of thickness, this thickness is straight
Connecing influences hydrophobicity quality, and hydrophobic layer is thin, and contact angle is small, and hydrophobicity is poor;Hydrophobic thickness, contact angle is big, and hydrophobicity is good.
Hydrophilic layer thickness 300nm-500nm, hydrophilic layer thickness are less than hydrophobic layer thickness, and hydrophilic region is fluted, works as sample
It drops on target spot, it is hydrophobic outside hydrophilic pores in hole that crystallization is contributed to be formed and grown.
Claims (8)
1. micro nano structure substrate is for Mass Spectrometer Method biomolecule or the purposes of biomarker sample, the wherein system of the substrate
Preparation Method includes:
(1) select the substrate strong but pliable in texture, flatness is good as substrate;
(2) substrate is handled with silane coupling agent:
(i) substrate is cleaned:Substrate is successively immersed into acetone, methanol aqueous solution, chloroform, after being respectively washed, takes out drying;
(ii) in a heated condition, concentrated acid and hydrogen peroxide are slowly added in the aqueous solution containing substrate, are carried out fully oxidized anti-
Ying Hou is cleaned multiple times in chloroform, ultra-pure water respectively;
(iii) it places the substrate in clean container, the silane coupler solution hydrolyzed in advance is added, and ammonia-catalyzed is added, directly
It is fully completed to Silanization reaction;
(iv) substrate is taken out, is respectively placed in ethyl alcohol, ultra-pure water, chloroform, is cleaned by ultrasonic;
(v) by the substrate after cleaning, the substrate of 120 ° of contact angle > is measured and selects, as qualified substrate;
(3) the hydrophily processing in point sample area:Modulated acid reagent is directly dropped in point sample area, the acid reagent pair is passed through
After surface is chemically modified, hydrophily cavernous structure is formed, is then cleaned by ultrasonic, it is final to obtain micro nano structure substrate;
Wherein,
Step (1) described substrate is the substrate of stainless steel strong but pliable in texture, that flatness is good, diamond, monocrystalline silicon, quartz crystal;
Step (2) described silane coupling agent is selected from vinyl silanes, amino silane, dimethyldichlorosilane;
Step (3) described acid reagent is selected from hydrofluoric acid, sulfuric acid, hydrogen bromide.
2. purposes according to claim 1, wherein preparation method include:
Step (4) laser beam marking Quick Response Code, the Quick Response Code, can by being associated with the source-information of the biological information of patient or sample
The corresponding information of substrate for identification.
3. purposes according to claim 1 or 2, wherein the substrate is the silicon chip or quartz substrate of single-sided polishing;Institute
150 ° of contact angle > is stated, to form the thickness of super hydrophobic surface or the hydrophobic surface in nanometer to micron dimension.
4. purposes according to claim 1 or 2, wherein point sample area, which are drawn, is divided into the suitable sample rooms MALDI-TOF specification
Rectangle draws and is less than 10 μm every precision;And by lithography mask version, close and distant water area is formed in substrate surface, and boundary is smart
Degree reaches micron level.
5. purposes according to claim 1 or 2 can use code reader or code reader to read wherein when carrying out Mass Spectrometer Method
Quick Response Code, and the source-information and corresponding mass spectrometric data of the corresponding patient's biological information of substrate or sample are inputted into electricity simultaneously
Brain is encoded, stored and is identified.
6. purposes according to claim 5, wherein the hydrophilic point sample area quantity on the substrate is 48,56,70,96,
384, hydrophilic area shape is selected from circle, square, triangle or polygon.
7. purposes according to claim 6, wherein the hydrophilic region size range is 100 × 100 μm to 1 × 1mm, institute
It is 20 × 30-83 × 125mm to state sizes of substrate.
8. purposes according to claim 1 or 2, wherein the biomolecule or biomarker are albumen or polypeptide sample
Product, nucleic acid samples.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810053331.XA CN108387634A (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710118754.0A CN106872562B (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
CN201810053331.XA CN108387634A (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710118754.0A Division CN106872562B (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108387634A true CN108387634A (en) | 2018-08-10 |
Family
ID=59169326
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810053331.XA Pending CN108387634A (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
CN201810052139.9A Pending CN108120762A (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
CN201710118754.0A Expired - Fee Related CN106872562B (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810052139.9A Pending CN108120762A (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
CN201710118754.0A Expired - Fee Related CN106872562B (en) | 2017-03-01 | 2017-03-01 | Mass spectrum substrate and preparation method and purposes |
Country Status (1)
Country | Link |
---|---|
CN (3) | CN108387634A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111386463A (en) * | 2018-08-30 | 2020-07-07 | 株式会社Lg化学 | Method for relative quantitative analysis of polymers using MALDI mass spectrometry |
Families Citing this family (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107167512A (en) * | 2017-06-29 | 2017-09-15 | 浙江和谱生物科技有限公司 | Disposable target plate for substance assistant laser desorpted ionisation mass spectrometry |
CN107267592B (en) * | 2017-07-04 | 2021-05-21 | 北京毅新博创生物科技有限公司 | Mass spectrum substrate target holder and instrument for biomarker detection |
CN107177689B (en) * | 2017-07-05 | 2021-01-22 | 北京毅新博创生物科技有限公司 | Universal chip for detecting protein and nucleic acid by time-of-flight mass spectrometry |
CN107192757B (en) * | 2017-07-05 | 2018-10-19 | 北京毅新博创生物科技有限公司 | A kind of dual-purpose detection kit of mass spectrum |
CN107515242B (en) * | 2017-08-04 | 2019-12-10 | 清华大学 | silicon-based gold nanometer bowl array chip and preparation method and application thereof |
CN108267500A (en) * | 2018-01-03 | 2018-07-10 | 北京毅新博创生物科技有限公司 | Mass spectrum substrate target holder detects the purposes of biological sample for BIOMARK |
US11442039B2 (en) * | 2018-02-09 | 2022-09-13 | Hamamatsu Photonics K.K. | Sample support body, production method for sample support body, and sample ionization method |
US20210028002A1 (en) * | 2018-02-09 | 2021-01-28 | Hamamatsu Photonics K.K. | Sample supporting body, method for ionizing sample, and mass spectrometry method |
JP6962831B2 (en) * | 2018-02-09 | 2021-11-05 | 浜松ホトニクス株式会社 | Ionization method and sample support |
CN109115685B (en) * | 2018-10-30 | 2021-02-09 | 上海交通大学 | Auxiliary positioning system for ultrathin section of electron microscope |
CN109541012A (en) * | 2018-11-23 | 2019-03-29 | 杭州汇健科技有限公司 | A kind of universal nano chips and the preparation method and application thereof for mass spectral analysis |
SG11202106282RA (en) * | 2018-12-12 | 2021-07-29 | Bgi Shenzhen | Biochip, method of preparation and use thereof |
CN109900774B (en) * | 2018-12-13 | 2020-07-28 | 中国科学院生态环境研究中心 | Method for improving detection sensitivity of MA L DI-TOF-MS to PS micro/nano particles |
CN109706066B (en) * | 2018-12-29 | 2022-08-26 | 赛纳生物科技(北京)有限公司 | Gene sequencing chip micro-pit surface modification method |
CN110887892B (en) * | 2019-12-23 | 2022-08-19 | 复旦大学 | Mass spectrum detection method for small amount of samples |
CN112162028A (en) * | 2020-09-29 | 2021-01-01 | 中国农业科学院农业质量标准与检测技术研究所 | Mass spectrum imaging method for vitamin C in strawberry tissue |
CN113008973A (en) * | 2021-01-29 | 2021-06-22 | 融智生物科技(青岛)有限公司 | Protein chip suitable for detecting low-abundance protein and preparation method and application thereof |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050072917A1 (en) * | 2003-09-30 | 2005-04-07 | Thomas Becker | Methods of making substrates for mass spectrometry analysis and related devices |
CN102168011A (en) * | 2010-12-31 | 2011-08-31 | 浙江大学 | PCR chip based on droplet array and application thereof |
CN103278618A (en) * | 2013-05-09 | 2013-09-04 | 董建国 | Biochip information reading device and information analyzing method |
CN104588137A (en) * | 2014-12-30 | 2015-05-06 | 厦门大学 | Micro-fluidic chip and preparation method thereof |
CN105324831A (en) * | 2013-03-13 | 2016-02-10 | 基纳生物技术有限公司 | Preparation enhancements and methods of use for MALDI mass spectrometry |
CN106085173A (en) * | 2016-06-12 | 2016-11-09 | 武汉理工大学 | A kind of functionally gradient composite construction abrasion-resistant clear super-hydrophobic coat and preparation method thereof |
CN106183420A (en) * | 2016-08-04 | 2016-12-07 | 纳晶科技股份有限公司 | A kind of fluid jetting head, for the nozzle plate of fluid jetting head and the manufacture method of this nozzle plate |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPWO2004051229A1 (en) * | 2002-12-02 | 2006-04-06 | 日本電気株式会社 | Liquid switch and microchip and mass spectrometry system using the same |
CN202230053U (en) * | 2011-07-28 | 2012-05-23 | 马庆伟 | Disposable target plate used for mass spectrometric detection |
CN102519779B (en) * | 2011-12-06 | 2013-06-12 | 吉林大学 | Concentration and demineralization purification treatment method of biological samples |
CN103234962A (en) * | 2013-05-09 | 2013-08-07 | 董建国 | Vaginal micro-ecological environment sensor and manufacturing method thereof |
CN103901093B (en) * | 2014-03-13 | 2017-01-04 | 华东理工大学 | Prepare the alternate micro-array chip of hydrophobe and the method for mass spectrum imaging quantitative analysis thereof |
CN104492676A (en) * | 2014-12-12 | 2015-04-08 | 哈尔滨工业大学 | Preparing method of polytetrafluoroethylene hydrophobic film |
CN105505742A (en) * | 2015-12-25 | 2016-04-20 | 中国科学院深圳先进技术研究院 | Drop array chip and preparation method thereof |
-
2017
- 2017-03-01 CN CN201810053331.XA patent/CN108387634A/en active Pending
- 2017-03-01 CN CN201810052139.9A patent/CN108120762A/en active Pending
- 2017-03-01 CN CN201710118754.0A patent/CN106872562B/en not_active Expired - Fee Related
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050072917A1 (en) * | 2003-09-30 | 2005-04-07 | Thomas Becker | Methods of making substrates for mass spectrometry analysis and related devices |
CN102168011A (en) * | 2010-12-31 | 2011-08-31 | 浙江大学 | PCR chip based on droplet array and application thereof |
CN105324831A (en) * | 2013-03-13 | 2016-02-10 | 基纳生物技术有限公司 | Preparation enhancements and methods of use for MALDI mass spectrometry |
CN103278618A (en) * | 2013-05-09 | 2013-09-04 | 董建国 | Biochip information reading device and information analyzing method |
CN104588137A (en) * | 2014-12-30 | 2015-05-06 | 厦门大学 | Micro-fluidic chip and preparation method thereof |
CN106085173A (en) * | 2016-06-12 | 2016-11-09 | 武汉理工大学 | A kind of functionally gradient composite construction abrasion-resistant clear super-hydrophobic coat and preparation method thereof |
CN106183420A (en) * | 2016-08-04 | 2016-12-07 | 纳晶科技股份有限公司 | A kind of fluid jetting head, for the nozzle plate of fluid jetting head and the manufacture method of this nozzle plate |
Non-Patent Citations (4)
Title |
---|
BEREND-JAN DE GANS ET AL.: "Inkjet Printing of Well-Defined Polymer Dots and Arrays", 《LANGMUIR》 * |
DONGHOON KWAK ET AL.: "Self-Organization of Inkjet-Printed Organic Semiconductor Films Prepared in Inkjet-Etched Microwells", 《ADV. FUNCT. MATER.》 * |
KOJI ABE ET AL.: "Inkjet-Printed Microfluidic Multianalyte Chemical Sensing Paper", 《ANAL.CHEM.》 * |
杨洪兴等: "《绿色建筑发展与可再生能源应用》", 31 December 2016, 中国铁道出版社 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111386463A (en) * | 2018-08-30 | 2020-07-07 | 株式会社Lg化学 | Method for relative quantitative analysis of polymers using MALDI mass spectrometry |
Also Published As
Publication number | Publication date |
---|---|
CN108120762A (en) | 2018-06-05 |
CN106872562B (en) | 2018-03-23 |
CN106872562A (en) | 2017-06-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN106872562B (en) | Mass spectrum substrate and preparation method and purposes | |
CN107192757B (en) | A kind of dual-purpose detection kit of mass spectrum | |
CN111733056B (en) | Micro-fluidic chip integrating circulating tumor cell separation and single-cell immunoblotting | |
CN103048462B (en) | Multi-parameter electrochemical immunosensor based on electrode array and preparation method thereof | |
KR102533281B1 (en) | General-purpose nanochip for mass spectrometry and method of manufacturing and using the same | |
CN107177689B (en) | Universal chip for detecting protein and nucleic acid by time-of-flight mass spectrometry | |
CN103335984B (en) | A kind of incorporeity wall micro-array chip based on LSPR and application thereof | |
CN110146463A (en) | A kind of method of multifrequency point resonant biosensor and preparation method thereof and test cell concentration | |
US20240094207A1 (en) | Serological biomarkers for early diagnosis of lung cancer | |
CN107179412B (en) | The preparation method of the general-purpose chip of albumen and nucleic acid is detected for flight time mass spectrum | |
CN202230053U (en) | Disposable target plate used for mass spectrometric detection | |
CN109632765A (en) | A kind of excretion body surface face method of protein detection | |
CN105992826B (en) | Method for nucleic acid analysis and equipment | |
US11280784B2 (en) | Patterned plasmonic nanoparticle arrays for multiplexed, microfluidic biosensing assays | |
CN111551713A (en) | COVID-19 virus antibody detection microsphere, preparation method thereof and kit containing microsphere | |
CN102507444B (en) | Auxiliary optical device of attenuation total reflection surface enhanced infrared spectrometer for DNA analysis | |
Khosravi et al. | Label-free capture of breast cancer cells spiked in buffy coats using carbon nanotube antibody micro-arrays | |
CN111504739A (en) | Immunohistochemical slide glass for detection | |
US20200348289A1 (en) | Sensor Device for Biosensing and Other Applications | |
WO2020069661A1 (en) | Serological biomarkers for early diagnosis of lung cancer | |
CN109370891A (en) | A kind of biochip and preparation method thereof | |
CN103760361A (en) | Biological chip for detecting person Hsp90a (heat shock protein 90) and detection method thereof | |
WO2022121625A1 (en) | Fluorescence detection chip, and preparation method therefor and use thereof | |
CN109939751A (en) | A kind of micro-fluidic chip of whole blood test, detection device and its detection method | |
Huang et al. | Rapid Detection of SARS-CoV-2 in Clinical and Environmental Samples via a Resonant Cavity SERS Platform within 20 min |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |