CN108593910A - Based on microsphere supported particle detection systems and method - Google Patents
Based on microsphere supported particle detection systems and method Download PDFInfo
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- CN108593910A CN108593910A CN201810306124.0A CN201810306124A CN108593910A CN 108593910 A CN108593910 A CN 108593910A CN 201810306124 A CN201810306124 A CN 201810306124A CN 108593910 A CN108593910 A CN 108593910A
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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Abstract
The present invention relates to a kind of based on microsphere supported particle detection systems and method, this system includes heating unit, sample bin chamber unit, wherein, the immune microsphere fluid for being combined with target biological molecules is mounted in the sample bin chamber unit, the immune microsphere is fixed on microsphere surface and is made for antibody or aptamer, the immune microsphere for being combined with target biological molecules uses fluorescent marker, after heating unit heats the sample bin chamber unit, thermophoretic effect is generated in the sample bin chamber unit, the excretion body is converged in into the lower side of temperature in the sample bin chamber unit, the indoor fluorescence signal marked on the immune microsphere of target biological molecules that is combined with of sample to be amplified, to be detected.The large biological molecules such as floating preteins of the present invention, nucleic acid modify the antibody or aptamer that can be combined with target protein, nucleic acid specificity in the spherome surface of micron-scale, obtain immune microsphere, are converged with the particle to free state, and be detected after convergence.
Description
Technical field
The present invention relates to micro-nano detection of particles fields more particularly to a kind of based on microsphere supported particle detection systems and side
Method.
Background technology
Micro-nano particle is detected in the prior art, to measure particle size, shape, concentration, activity etc., in blood
The subjects such as, immunology, molecular biology have relatively broad application.In the prior art frequently with streaming particle detection method pair
Micro-nano particle is detected, and is the technology that the fine-grained particles in liquid are carried out with quantitative analysis and sorting one by one, is being examined
Coulter principle employed in survey refers to：When the particle to suspend in the electrolytic solution passes through aperture with electrolyte, replace same volume
Long-pending electrolyte causes two resistance between electrode inside and outside aperture that instantaneous variation occurs in the circuit of constant current design, generates current potential
Pulse, the size and number of pulse signal are directly proportional to the size and number of particle.Sample focusing is the pass of streaming detection of particulates
Key technology is all to realize to focus to sample liquid by outer force effect in current detection.Focusing be divided into for by sheath fluid focus and
Focusing without sheath fluid.
Wherein, sheath fluid is focused as disclosed in Chinese patent 201210482142.7《Microfluid particle instrument and production method》
In, sample liquid is injected from sample liquid entrance respectively using the pressure of extraneous syringe pump, injects sheath fluid from sheath fluid entrance, then sample
Liquid and two-way sheath fluid flow to sheath stream assembling area simultaneously, and the fine-grained particles in sample liquid are sandwiched linear row by the aggtegation of sheath fluid
Row flow into detection zone and are detected.Two sheath streams and sample liquid are required for driving source in this method, are controlled using a motor
The mode of three pipelines, not only equipment becomes very huge in this way, and cost also improves, and what is more important due to being detected every time
When need replacing chip, then every time detection be required for again being attached in three channels with motor, this junction
Sealing problem just influences whether the size of the pressure to three channels, causes focusing effect bad, and test result is just not smart enough
Really.
Wherein, it is not necessarily to the focusing of sheath fluid, as disclosed in Chinese patent 201310283051.5《One kind being used for streaming particle instrument
Microfluidic chip structure and preparation method thereof》, use taper focusing structure, it is believed that it is with similar with traditional sheath liquid stream
The focusing effect of system so that fine-grained particles individually flow into microchannel, and microchannel fetters particle by channel makes it individually pass through
Detection zone causes the inaccurate of testing result under the testing conditions of high concentration sample.
In the technical solution that above two detects micro-nano particle, on the one hand, potential pulse is generated by electrolysis, using electricity
Chemical method is detached and is detected to nano particle, and a fluid stream of the particle containing micro-nano is formed, and the amount of required sample is very big；Another party
Face, by the driving source of such as motor, while using the single channel of fixed structure, limit micro-nano particle flow direction and
Direction is built up, during applying outer active force and channel limits, external force acts on fluid, is often directed to micro-nano particle
Force be uncontrollable.
Especially for micro-nano biomone, such as detection of excretion body, excretion body is film bubble secreted by cell, for thin
Intercellular exchange, because its contain with the relevant albumen of mother cell and inhereditary material, be increasingly becoming a kind of emerging non-intruding in recent years
The biomarker of formula diagnosing tumor.
Generally use in the prior art：First, enzyme linked immunosorbent assay (ELISA), i.e. ELISA, refer to soluble antigen or resist
Body is attached on the solid phase carriers such as polystyrene, and the qualitative and quantitative inspection of immune response is carried out using antigen-antibody binding specificity
Survey method, when measuring, by examining sample (measuring antibody or antigen therein) and enzyme-labelled antigen or antibody by different steps
It reacts with the antigen or antibody of surface of solid phase carriers；Make the antigen antibody complex formed on solid phase carrier with the method for washing
It is separated with other substances, finally combines the amount of tested substance in enzyme amount and the sample on solid phase carrier at certain ratio.Add
After the substrate for entering enzyme reaction, substrate becomes color products, the direct phase of amount of the amount of product and tested substance in sample by enzymatic
It closes, therefore can be according to the depth publication qualitative or quantitative analysis of color reaction.
Second, immunoblotting, i.e., Western Blot, basic principle are by specific antibody to gel electrophoresis
Processed cell or biological tissue samples are coloured；Position by analyzing coloring obtains specific protein with color depth
The information of expression in the cell or tissue analyzed.
On the one hand above two technical solution is detached and is built up to sample, special device and method need to be used；Separately
On the one hand, detection method needs to use a large amount of sample, is often directed to the process that serum carries out canceration detection and does not have adaptability.
European Patent Publication No EP2783747 also discloses a kind of liquid mixed method using to hot-fluid, but it passes through
Liquid is set to pass through in capillary, also, to be positioned over gravity parallel and/or anti-parallel guide for capillary, due in capillary
Liquid is guided, liquid cannot be moved to specified same direction in particle, cannot be built up, it is thus impossible to pass through the bootstrap technique
Carry out the detection of particulate.It cannot be gathered especially for the particle of free state, such as floating preteins, nucleic acid large biological molecule
Product, can not realize the detection to particle.
Invention content
The purpose of the present invention is to provide a kind of based on microsphere supported particle detection systems and method, existing to overcome
It cannot be to the accumulation detection technique defect of the particle of free state in technology.
To achieve the above object, the present invention provides a kind of based on microsphere supported particle detection systems, including heating unit,
Sample bin chamber unit, wherein
The heating unit, to the sample heating into the sample bin chamber unit；
The immune microsphere fluid for being combined with target biological molecules, the immune microsphere are mounted in the sample bin chamber unit
Microsphere surface is fixed on for antibody or aptamer to be made, the immune microsphere for being combined with target biological molecules uses fluorescence mark
Note generates thermophoretic effect, by institute after the heating unit heats the sample bin chamber unit in the sample bin chamber unit
It states excretion body and converges in the lower side of temperature in the sample bin chamber unit, the indoor target organism that is combined with of sample is divided
The fluorescence signal amplification marked on the immune microsphere of son, to be detected.
Further, the system also includes signal gathering unit, the signal gathering unit obtains amplified fluorescence
Signal obtains the abundance for the target biological molecules that immune microsphere surface is combined, by by various aptamers respectively be combined with mesh
The different target proteins combination for marking the immune microsphere surface of biomolecule obtains testing result.
Further, the sample bin chamber unit includes loading to be combined with the immune microsphere of target biological molecules and to carry
For generating the closed sample room in thermophoretic effect space, the sample room includes：To close the sample room and build up described micro-
Receive the second thermal conductive surface of particle.
Further, nearby temperature is less than the temperature of the micro-nano particle fluid to second thermal conductive surface, with described the
The temperature difference is generated between two thermal conductive surfaces and the immune microsphere fluid for being combined with target biological molecules, is generated thermophoretic effect, is driven
The immune microspheres of target biological molecules is combined with to the second thermal conductive surface displacement.
Further, the heating unit is the laser being arranged on the outside of the sample bin chamber unit, to the sample
Product warehouse unit irradiates, and light beam passes sequentially through the immune microsphere fluid and the second thermal conductive surface for being combined with target biological molecules,
To generate thermophoretic effect to the immune microsphere solution for being combined with target biological molecules.
Further, the sample room further includes：To close the first thermal conductive surface of the sample room, second heat conduction
Face and the first thermal conductive surface can be such that light beam passes through.
Further, second thermal conductive surface is transparent material, is sapphire material；
First thermal conductive surface is any one of glass, polymethyl methacrylate, dimethyl silicone polymer, sapphire
Or appoint several combinations.
Further, the microballoon is polystyrene microsphere.
The present invention also provides a kind of based on microsphere supported particle detecting method, including：
Immune microsphere is prepared, microballoon and antibody or aptamer are incubated jointly, so that antibody or aptamer is fixed on microsphere surface, obtains
To immune microsphere；
Immune microsphere and detected sample are incubated, the target protein or nucleic acid in detected sample and immune microsphere
On antibody or aptamer specifically bound, to be fixed on immune microsphere；
By the immune microsphere obtained above for being combined with target biological molecules and the antibody or aptamer knot that carry fluorophor
It closes, by the target biological molecules mark fluorescent on immune microsphere；
The immune microsphere sample for being combined with target biological molecules loaded in sample bin chamber unit is heated, in sample
Warehouse unit generates thermophoretic effect, and the immune microsphere for being combined with target biological molecules is converged in the sample bin chamber unit
Low temperature side, and due to fluorescent marker be enriched with and so that signal is amplified, to be detected.
Further, target biological molecules are combined with by what is built up to the low temperature side in the sample bin chamber unit
Immune microsphere, acquisition are combined with the immune microsphere corresponding index information of target biological molecules and analyze corresponding index.
The beneficial effects of the present invention are micro-nano detecting system of the present invention passes through to micro-nano particle institute compared with prior art
The wherein direction of sample bin chamber unit heated, introduce thermophoretic effect and convection current, sample bin chamber unit made to generate temperature
Difference generates low temperature in the side far from heating unit, and thermophoretic effect makes micro-nano particle migration in sample and is accumulated to sample warehouse
Unit, to complete the accumulation of micro-nano particle；Simultaneously as sample liquids expanded by heating generates buoyancy in sample bin chamber unit
Middle generation convection current, in the low-temperature region of sample bin chamber unit, the heating zone of sample bin chamber unit is directed toward in the direction of convection current from surrounding
Domain further promotes the accumulation of micro-nano particle.
In particular, being not exposed to albumen, the core on surface in the large biological molecules such as floating preteins of the present invention, nucleic acid or excretion body
The large biological molecules such as acid, the antibody that can be combined with target protein, nucleic acid specificity in the spherome surface modification of micron-scale or suitable
Body, obtains immune microsphere, and it with the sample incubation containing target biomacromolecule and is combined simultaneously with target biomacromolecule
Mark fluorescent, with to free state particle or be not exposed to the target biomacromolecule on surface and converge, and it is laggard converging
Row detection.
Further, this system passes through the aptamer or antibody incubation of the sample to be tested containing excretion body and fluorescent marker
Aptamer or antibody are combined with excretion body surface face protein-specific marks upper fluorescence by excretion body；The sample being incubated is put into up and down
Transparent sample room, and be placed on fluorescence microscope objective table and be observed, infrared laser shines through sample room effect
With sample, acted on by thermophoresis excretion body in sample is highly enriched at the laser spot of sample room bottom so that excretion body
Fluorescence is highly enlarged, and the abundance of certain excretion body surface protein is detected by fluorescence power.
Further, this system heats sample room irradiation using laser, by being arranged in sample room opposite flank
The different transparent thermal conductive surface of heat conductivility, the temperature difference is generated between two thermal conductive surfaces, to generate thermophoretic effect, drives micro-nano particle
From the lower second thermal conductive surface displacement of the first thermal conductive surface low temperature.It is heated especially with light beam, other auxiliary need not be used
Equipment only needs sample room transparent thermal conductive surface setting up and down.Also, micro-nano particle is straight in thermophoretic effect lower stress and particle
Diameter square is directly proportional, and how much unrelated with micro-nano number of particles, therefore, only needs micro micro-nano particle that convergence and inspection can be completed
Survey, 0.1 microlitre of sample dosage only needed for excretion body, it is easy to operate, be not necessarily to specific apparatus, and without sample pre-treatments and
Excretion body purifies, and is common to aptamer and antibody；It is not limited to excretion body, the micro-nanos biomone such as other extracellular vesica, cells is equal
It can.
Especially, present system and method can be selected to complete to measure under specific temperature, not limited by actual temp
It is fixed, it only needs that the temperature difference can be generated to build up particle；It can also complete to measure in a variety of different solution environmentals, including research
Complicated detergent environment needed for memebrane protein；It can also be to various different size of molecules：As ion, nucleic acid fragment, nucleosome,
Liposome is detected, and in specific be detected, system can be adjusted according to the physical property of particle itself and the size of particle
The parameters such as the type of height, fluid between the temperature difference, upper and lower thermal conductive surface and the frequency of laser irradiation are adjusted, above-mentioned each ginseng
The adjustment of amount can realize quantitative adjustment, and control is accurate, easy to adjust.
The present invention is based on thermophoretic effect build up particle, the loading container of micro-nano particle is not limited, especially volume compared with
In big container, it is easier to which particle is built up under thermophoretic effect, is guided without being considered as the carrier containers such as capillary.
Description of the drawings
Fig. 1 is the structure diagram of the micro-nano particle detection systems of the present invention；
Fig. 2 is the signal detection flow diagram based on excretion body of the present invention；
Fig. 3 is the comparison collection of illustrative plates of the excretion body of the embodiment of the present invention 1 before and after the test；
Fig. 4 is excretion body each surface protein collection of illustrative plates after sensing of the embodiment of the present invention 2 and corresponding each surface protein
Expression quantity schematic diagram；
Fig. 5 is 2 all kinds of cancer patients of the embodiment of the present invention in all kinds of expressing quantities of the serum excretion body of Healthy People
Schematic diagram；
Fig. 6 is the fluorescence measurement gray value schematic diagram of the fluorescence polystyrene microsphere of the different-diameter of the embodiment of the present invention 3；
Fig. 7 is the ovarian cancer patients of the embodiment of the present invention 4 and showing for 11 kinds of albumen marker expression amounts in Healthy Human Serum
It is intended to；
Fig. 8 be the embodiment of the present invention 4 11 unlike signal objects and its summation as distinguishing the correct of cancer and health standards
Rate schematic diagram.
Specific implementation mode
Below in conjunction with attached drawing, the forgoing and additional technical features and advantages are described in more detail.
The preferred embodiment of the present invention described with reference to the accompanying drawings.It will be apparent to a skilled person that this
A little embodiments are used only for explaining the technical principle of the present invention, it is not intended that limit the scope of the invention.
It should be noted that in the description of the present invention, the instructions such as term "upper", "lower", "left", "right", "inner", "outside"
Direction or the term of position relationship be direction based on ... shown in the drawings or position relationship, this is intended merely to facilitate description, and
It is not instruction or implies that described device or element must have a particular orientation, with specific azimuth configuration and operation, therefore not
It can be interpreted as limitation of the present invention.In addition, term " first ", " second " are used for description purposes only, and should not be understood as indicating
Or imply relative importance.
In addition it is also necessary to explanation, in the description of the present invention unless specifically defined or limited otherwise, term " peace
Dress ", " connected ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or integrally
Connection；It can be mechanical connection, can also be electrical connection；Can be directly connected, can also indirectly connected through an intermediary,
It can be the connection inside two elements.To those skilled in the art, it can understand that above-mentioned term exists as the case may be
Concrete meaning in the present invention.
Refering to Figure 1, its structure diagram for the micro-nano particle detection systems of the present invention, the system packet of the present embodiment
Include heating unit 1, sample bin chamber unit 2, signal gathering unit 4, wherein the heating unit 1 is arranged in the sample warehouse
The outside of unit 2, to the sample heating into the sample bin chamber unit 2；It is mounted with micro-nano in the sample bin chamber unit 2
Particle generates thermophoresis effect after the heating unit 1 heats the sample bin chamber unit 2, in the sample bin chamber unit 2
It answers, micro-nano particle is converged in the side far from the heating unit 1 in the sample bin chamber unit 2；The signal acquisition
Unit 4 after the micro-nano particle in the sample bin chamber unit 2 is built up, acquires the coherent signal letter of the micro-nano particle
Breath, and corresponding analysis is carried out to corresponding micro-nano particle.This system utilizes thermophoretic effect, i.e., object is under temperature gradient effect
Directional migration is radiated at sample by infrared laser and locally generates temperature gradient field, the excretion body in sample is made to move to temperature
Lower.It is heated by the wherein direction to the sample bin chamber unit 2 where micro-nano particle, introduces thermophoretic effect and right
Stream makes the side that sample bin chamber unit 2 generates the micro-nano particle fluid in the temperature difference and sample bin chamber unit 2 generate the low temperature temperature difference,
And the temperature of the side of sample bin chamber unit 2 is made to be less than the temperature of micro-nano particle fluid, thermophoretic effect makes micro-nano particle in sample
Migrate and be accumulated to the low temperature side of sample bin chamber unit 2；Simultaneously as sample liquids fluid expanded by heating generate buoyancy to
Convection current is generated in sample bin chamber unit 2.In the direction of the low-temperature region of sample bin chamber unit 2, convection current sample is directed toward from surrounding
The heating region of warehouse unit 2 plays the role of conveyer belt and micro-nano particle around is converged in sample such as arrow direction in Fig. 1
The 2 low temperature side of sample bin chamber unit of product warehouse unit 2, to play the role of building up micro-nano particle.Therefore, the present invention is based on
Thermophoretic effect builds up particle, does not limit the loading container of micro-nano particle, especially in the larger container of volume, it is easier to grain
Son is built up under thermophoretic effect, is guided without being considered as the carrier containers such as capillary.
Specifically, the heating unit 1 of the present embodiment is laser, it is arranged in the outside of sample bin chamber unit 2, to sample
Be irradiated in product warehouse unit 2, to generate circular heating region inside it, certain heating region may be it is linear or
Person's other manner.It will be appreciated by persons skilled in the art that mode of heating is not limited in laser irradiation, laser irradiation direction
It only needs to ensure to generate heat source, the selection of power depends on direction of illumination, and the factors such as spot diameter, wavelength can root
Change according to actual micro-nano particle and use environment.
Specifically, sample bin chamber unit 2 includes loading micro-nano particle samples and generating thermophoretic effect space to provide
Closed sample room 24, sample room 24 includes closing the first thermal conductive surface 21 of sample room 24, and to close sample room
24 the second thermal conductive surface 22 produces between micro-nano particle fluid and the second thermal conductive surface 22 that sample room 24 loads in the present embodiment
The raw temperature difference drives micro-nano particle from micro-nano particle fluid to 22 displacement of the second thermal conductive surface to generate thermophoretic effect.Therefore,
Temperature near second thermal conductive surface 22 is less than the temperature of micro-nano particle fluid by the present embodiment.
In the present embodiment, sample room 24 is heated using laser, first thermal conductive surface 21, the second thermal conductive surface
22 are oppositely arranged, and the thermal conductivity of second thermal conductive surface 22 is more than the first thermal conductive surface 21, and the heat dissipation performance of the second thermal conductive surface 22 is big
In the first thermal conductive surface 21, and two thermal conductive surfaces are transparent material, convenient for being observed to micro-nano particle, therefore, the second heat conduction
The temperature in face 22 is less than the temperature of the first thermal conductive surface 21, but also the temperature of micro-nano particle fluid is less than the temperature of the second thermal conductive surface 22
Degree.The sample room 24 further includes the gasket 23 to sealed sample room 24, it will be appreciated by persons skilled in the art that two
Thermal conductive surface 21 can be oppositely arranged or be disposed adjacent, or be arranged between each other according to default angle, and micro-nano particle edge need to only be driven to set
Fixed a direction is moved, is built up.The fluid of the present embodiment can be liquid, the mixed liquor of such as water or water, or
Gas only needs that micro-nano particle can be loaded such as heat gas or natural mode gas, and can allow for micro-nano particle in a fluid from
By moving, to be built up using thermophoretic effect.Meanwhile first thermal conductive surface 21 and the second thermal conductive surface 22 be it is transparent, can be with
The first thermal conductive surface and the second thermal conductive surface are sequentially passed through by infrared ray, and heat is brought into the fluid, naturally it is also possible to be
Electromagnetic radiation generates heat, so that sample room generates the temperature difference.
As preferably embodiment, the first thermal conductive surface 21 is glass, polymethyl methacrylate (PMMA), poly dimethyl silicon
Oxygen alkane (PDMS), sapphire etc., the second thermal conductive surface 22 are the good sapphire of thermal conductivity or diamond.Laser is by sequentially irradiating
One thermal conductive surface 21, the sample room 24 for loading micro-nano particle, the second thermal conductive surface 22 generate low-temperature space in the second thermal conductive surface 22.It will swash
Optical focus is adjusted in sample room 24, and laser absorbs laser by the sample liquids in region in sample room 24 and temperature increases, heat
Swimming effect makes in sample micro-nano particle migration arrive lower second thermal conductive surface 22 of temperature, simultaneously because the production of sample liquids expanded by heating
Capture power in sample room so that generate convection current；Low temperature direction near the second thermal conductive surface 22, the direction of convection current refer to from surrounding
To illuminated laser spot, plays the role of conveyer belt and micro-nano particle around is converged in the second heat conduction of sample room below illuminated laser spot
The region in face 22 is built up to enhance micro-nano particle.
In a wherein embodiment of the invention, micro-nano particle selection is excretion body, and excretion body is film bubble secreted by cell,
For intercellular communication, because its contain with the relevant albumen of mother cell and inhereditary material, be increasingly becoming in recent years a kind of emerging
The biomarker of non-intrusion type diagnosing tumor.Concrete principle of the present embodiment based on excretion body is as described below.
Excretion body heat swimming motion model：
vT=-STD▽T (1)
Wherein, vTFor thermophoresis speed, STFor Soret coefficients, D is diffusion coefficient, and ▽ T are temperature gradient, model formation right end
Negative sign indicates that thermophoresis direction is low temperature direction.
Soret coefficient formulas in above-mentioned formula (1)：
Wherein, A is excretion body surface area, and k is Boltzmann constant, and T is temperature, shydFor aquation entropy, β is coefficient,
σeffFor the equivalent charge density in excretion body surface face, λDHFor Debye length, ε0For permittivity of vacuum, ε relative dielectric constants.It is comprehensive
Above-mentioned formula (1)-(2), it is known that excretion body heat swimming stress is directly proportional to diameter square.
Excretion body migration models in thermal convection current：
Res=ρ a | u-Vp|/η (5)
Wherein, VpFor movement velocity of the excretion body under thermal convection current effect, a is excretion body diameter, and u is thermal convection current speed, CD
For viscosity coefficient, can be calculated according to formula (4), wherein a1、a2、a3For constant, ResIt, can be according to formula for relative motion Reynolds number
(5) it calculates, g is acceleration of gravity, ρpFor excretion body averag density, ρ is sample liquids density.Aggregative formula (3)-(5), it is known that
The viscous resistance of excretion body thermal convection is directly proportional to diameter.
Compare thermophoretic forces and thermal convection current viscous resistance, it is known that object is bigger, and the thermophoretic forces being subject to more account for leading, more tend to
The second thermal conductive surface being gathered in sample room bottom surface namely the present embodiment device；Object is smaller, is got over by thermal convection current viscous resistance
It accounts for leading, more tends to follow thermal convection current without assembling.Therefore, the present invention makes the micro-nano particle in liquid by thermophoretic effect
It flows and assembles.Certainly, above-mentioned model is suitable for all micro-nano particles, does not also limit the grain size of micro-nano particle.
Shown in Fig. 1, signal amplification unit 3 includes that the micro-nano particle that is arranged in the sample bin chamber unit 2 is poly-
The microscope in product region comprising be directed at the object lens 31, reflective mirror 32 and observation light source 33 of the micro-nano particle of accumulation, pass through
Microscope more can clearly observe micro-nano particle.The signal gathering unit 4 be CCD camera, it is of course also possible to be it is any can
The instrument for detecting optical signal, takes pictures to micro-nano particle by microscope, obtains information.
In the present embodiment, for excretion body signal detection, the aptamer of excretion body sample and fluorescent marker is incubated first
It educates, the aptamer for making fluorescent marker and the target protein in excretion body surface face are specifically bound, to which excretion body is marked upper fluorescence；It will incubate
Excretion body sample after educating is put into sample room 24, and is heated by laser and introduce thermophoretic effect and convection current, and sample is indoor outer
Secrete the fluorescence signal amplification marked on body；By the fluorescence signal after CCD recording laser pre-irradiations, before analyzing laser irradiation
Fluorescence signal afterwards obtains the abundance of excretion body surface face target protein；It, can using a series of aptamers that can combine different target proteins
It obtains excretion body surface face protein graphical spectrum, and finally determines the corresponding index parameter of excretion body by the analysis.
In the present embodiment, the detection method of micro-nano particle includes：
Step a heats the micro-nano particle samples in sample bin chamber unit 2 from side, is produced in sample bin chamber unit 2
Raw thermophoretic effect, the low temperature side in the sample bin chamber unit 2 is converged in by micro-nano particle；
In above-mentioned steps a, sample fluid expanded by heating generates buoyancy to generating convection current in sample bin chamber unit 2,
In the low-temperature region of sample bin chamber unit 2, the heating region of sample bin chamber unit 2 is directed toward in the direction of convection current from surrounding, by surrounding
Micro-nano particle converges in the low temperature side of sample bin chamber unit 2.
Step b acquires micro-nano particle by the micro-nano particle built up to the low temperature side in the sample bin chamber unit 2
Corresponding index information and corresponding index is analyzed.
Specifically, the present embodiment carries out signal detection to excretion body, in conjunction with shown in Fig. 2, which is：
The aptamer of excretion body sample and fluorescent marker is incubated by step a1, makes aptamer and the excretion body surface face of fluorescent marker
Target protein specific binding, to which excretion body is marked upper fluorescence；
Excretion body sample after incubation is put into sample room by step a2, and is heated by laser and to be introduced thermophoretic effect and right
Stream, the indoor low temperature side of the sample, the fluorescence signal that will be marked on the indoor excretion body of sample are converged in by excretion body
Amplification；
Step a3 obtains the front and back fluorescence signal of light irradiation and is obtained out by the fluorescence signal before and after analysis laser irradiation
Secrete the abundance of body surface face target protein；
Step a4 obtains excretion body surface face protein graphical spectrum using a series of aptamers that can combine different target proteins.
Above-mentioned micro-nano particle detection systems and method are illustrated below by specific embodiment.
Embodiment 1
The aptamer of excretion body sample and fluorescent marker is incubated, the aptamer of selection is through in-vitro screening technology SELEX (indexes
Enrichment Fas lignand system is evolved) oligonucleotide fragment of energy specific binding protein or other small-molecule substances that filters out, tool
For body, the aptamer of fluorescent marker is the single stranded DNA of 40 bases, and the ball of string diameter in sample liquids is less than 5 nanometers, and excretion
Body is 30-150 nanometers a diameter of；The aptamer of specific recognition CD63 albumen is trained applied to A375 cells (human melanoma cell)
Support the excretion body in base supernatant.Fluorophor can be modified in aptamer end by standard approach, when aptamer and excretion body surface face
When specificity between target protein interacts, the fluorescence of aptamer carrying is marked in excretion body.The excretion body sample of the present embodiment
Incubation conditions for cell culture medium supernatant, sample are：2 hours incubation times, are incubated 0.1 every liter of micromole of aptamer concentrations
Temperature room temperature.
Wherein, laser is heated using the infrared laser of 1480nm wavelength for sample, and power is 200 milliwatts, and focus goes out
About 200 microns of laser spot diameter.Because the general main component of sample liquids is water, water is to there are one absorb near 1480nm wave bands
Peak, it will be appreciated by persons skilled in the art that mode of heating is not limited in laser irradiation, wavelength is also not limited to
1480nm, laser irradiation direction are not limited to irradiate from the top down, and the selection of power depends on direction of illumination, spot diameter, wavelength
Etc. factors, be not limited to 200 milliwatts.In the present embodiment, laser irradiates from top to bottom, and the upper thermal conductive surface of sample room uses bright material
Matter, such as glass, PMMA, PDMS, lower thermal conductive surface uses the better sapphire of thermal conductivity, and forming low-temperature space in bottom surface makes excretion body heat
Swimming converges at bottom surface.The thickness of upper thermal conductive surface is 1mm, and the thickness of lower thermal conductive surface is the height of 1mm, intermediate washer and sample room
It is 240mm.
Operated according to the above-mentioned signal detecting method based on excretion body, when aptamer identify excretion body surface protein and with
Combination when, the fluorescent marker on aptamer follows the sample room bottom section that excretion body is accumulated below laser spot, and produces
Raw enhancing fluorescence signal；When the unidentified excretion body surface protein of aptamer, free aptamer cannot be converged since size is small, signal
Do not enhance.In conjunction with shown in Fig. 3, in the present embodiment, CD63 albumen is widely present in the excretion body surface face of various types of cells, passes through laser
After irradiation, there is apparent fluorescence signal, shows that the excretion body surface face of A375 cells has CD63 albumen.
The fluorescence signal marked on the aptamer after being combined with excretion body is excited and received using fluorescence microscope, is excited and is connect
The wavelength for receiving fluorescence is related to the fluorescence radiation group properties of label, in the present embodiment, luminophore Cy5 excitation/emission wavelength
For 649/666nm, the CCD records that fluorescence signal is connect with fluorescence microscope.Pass through the fluorescence after CCD recording laser pre-irradiations
Signal obtains the abundance of excretion body surface face target protein by the fluorescence signal before and after analysis laser irradiation.
Embodiment 2
The present embodiment, using cervical cancer patient blood serum sample, using 7 kinds of different aptamers to excretion body 7 in blood serum sample
The abundance of kind surface protein (CD63, PTK7, EpCAM, HepG2, HER2, PSA, CA125) is detected, and and Healthy Human Serum
Sample is compared.
The excretion body operating method and laser of use, sample room and microscope and CCD camera all same.
In conjunction with shown in Fig. 4, it is known that this cervical cancer patient serum excretion body height expresses CD63 albumen and cancer Research of predicting markers
PTK7, EpCAM, HepG2, HER2, PSA and CA125, wherein CA125 can be used as the marker of traditional cervical carcinoma, also there is portion
Dividing cervical cancer patient, there are HER2 high expression.It is generally acknowledged that tumor markers PTK7, EpCAM are related to kinds cancer, HepG2 master
To be directed to liver cancer has specificity, PSA to have specificity mainly for prostate cancer.But these tumor markers are not and certain cancer
Disease has stringent correspondence correlativity.But since tumour is during growth or cancer are to other organ metastasis, through excessive
Secondary division growth, cell constantly generate gene mutation, show the change in terms of molecular biology or gene but, so these are swollen
Tumor markers are not to have stringent corresponding correlativity with certain cancer.It is examined in this cervical cancer patient serum in the present embodiment
PTK7, EpCAM, HepG2 are measured, the potentiality of gene mutation or transfer of the method in capture tumour are embodied.In addition CD63 makees
For the albumen that excretion body is generally expressed, cancer patient's excretion body expression also above Healthy People, with existing traditional detection side
The result that method obtains is consistent.
This method is further applied into a large amount of true clinical blood serum samples, including 3 cervical carcinomas, 2 oophoromas, 2
Lymph cancer, 2 breast cancer and 2 Healthy Peoples.In conjunction with shown in Fig. 5, this method is capable of detecting when all kinds of cancer patients and health
The difference of all kinds of expressing quantities of the serum excretion body of people.Between variety classes cancer, serum excretion body protein expression quantity
Difference, be mainly manifested in HER2 expressed in breast cancer and cervical carcinoma it is higher, CA125 expressed in oophoroma and cervical carcinoma compared with
Height, PSA are not expressed in the cancer detected in type, and EpCAM and PTK7 and CD63 have higher table in kinds cancer
It reaches.These results are consistent with existing method testing result.
Illustrate that this method can delicately detect cancer patient's serum and the excretion body surface protein in Healthy Human Serum,
The difference of expression quantity including cancer markers.And show detection method using excretion body as cancer marker more
It is convenient, sensitive, effective：The tumor marker species of conventional cancer screening or physical examination detection are limited (to be limited to available expensive anti-
Body and reagent) and sensitivity it is not high and lead to false negative, i.e., marker is not detected in patient, for example, cervical carcinoma in the present embodiment
CA125 expressions of results are in normal range value in Venous Blood examining report.And this method is not necessarily to expensive antibody, according to inspection
It surveys and needs that the aptamer that can be combined with the protein-specific of corresponding tumor markers can be used.
Embodiment 3
The present embodiment, the micro-nano particle used are for abiotic micro-nano particle, specially fluorescence polystyrene microsphere, brand
Thermofisher, a diameter of 50 to 200 nanometers, mass fraction 0.001% is dissolved in containing the water-soluble of 0.02% Tween20
In liquid.Laser, sample room and microscope and CCD camera are identical as above-described embodiment 1,2.
In conjunction with shown in lower Fig. 5, the fluorescent microsphere of all different-diameters highly converges at laser spot, and according to fluorescence
Measure gray value convergence degree visible with fluorescence picture enhances as particle diameter increases with fluorescence intensity, with the present embodiment
Operation principle is consistent, i.e., bulky grain is more likely to converge.This example demonstrates that either biological or abiotic micro-nano particle,
It is suitable for the design of the technical program.
Embodiment 4
The present embodiment micro-nano particle is the egg that the large biological molecules such as floating preteins, nucleic acid or excretion body are not exposed to surface
In vain, the large biological molecules such as nucleic acid cannot directly build up the large biological molecule of free state using the thermophoretic effect of the various embodiments described above,
Therefore, the mechanism of the present embodiment is, can be combined with target protein, nucleic acid specificity in the spherome surface modification of micron-scale
Antibody or aptamer obtain immune microsphere, and divide greatly by it with the sample incubation containing target biomacromolecule and with target organism
Son combines and mark fluorescent.It is acted on by above-mentioned thermophoresis and converges microballoon height so that target biomacromolecule fluorescence signal is high
Degree amplification, and its abundance is detected by fluorescence power.
The present embodiment includes based on microsphere supported particle detecting method：
Step a11, prepares immune microsphere, and microballoon and antibody or aptamer are incubated jointly, so that antibody or aptamer is fixed on micro-
Ball surface obtains immune microsphere；In this process, the extra antibody or aptamer not combined with microballoon is washed away；In this implementation
In example, microballoon uses polystyrene microsphere.
Step b11, immune microsphere and detected sample are incubated, the target protein or nucleic acid in detected sample with
Antibody or aptamer on immune microsphere are specifically bound, to be fixed on immune microsphere；
Step c11 will be combined with the immune microsphere and carrying fluorophor of target biological molecules made from above-mentioned steps b11
Antibody or aptamer combine, by specific recognition, by the target biological molecules mark fluorescent on immune microsphere；
Step d11 carries out the immune microsphere sample for being combined with target biological molecules in sample bin chamber unit 2 from side
Heating generates thermophoretic effect in sample bin chamber unit 2, the immune microsphere for being combined with target biological molecules is converged in the sample
Low temperature side in product warehouse unit 2, and so that signal is amplified since fluorescent marker is enriched with；In this process, by generating heat
Swimming, since target biological molecules are captured by immune microsphere to which equivalent dimension becomes larger, by the amplification of highly enriched and signal rather than mesh
Mark biomolecule, which is in free state equivalent dimension very little signal, to be amplified.
Step e11 is combined with target biological molecules by what is built up to the low temperature side in the sample bin chamber unit 2
Immune microsphere, acquisition are combined with the corresponding index information of the immune microsphere of target biological molecules and analyze corresponding index.
In this process, it obtains light irradiation front and back fluorescence signal and excretion body is obtained by the fluorescence signal before and after analysis laser irradiation
The abundance of surface target protein；Using a series of aptamers that can combine different target proteins, excretion body surface face protein graphical spectrum is obtained.
This example captures the protein marker that dissociates in ovarian cancer patient whole blood using the immune microsphere of pan coating antibody,
Thermophoresis is generated using infrared laser, detection is amplified to protein marker fluorescence signal, and determines the rich of testing protein marker
Degree, result meet with traditional detection method result, and molecular information is provided for cancer detection.In the present embodiment, for ovary
Cancer chooses EpCAM, CA-125, CA19-9, CD24, HER2, MUC18, EGFR, CLDN3, CD45, CD41, D2-40 as albumen
Immune microsphere is respectively prepared in the corresponding specific antibody of these protein markers (being bought from abcam companies) by marker, each
Microballoon is prepared separately in antibody, specifically for a kind of detection of marker.There is normal stream for the preparation of coated antibody immune microsphere
Journey can refer to, herein briefly narration：The antibody of 1 micron polystyrene microspheres of diameter and 5 μ g/ml concentration is incubated at room temperature 1
Hour, after incubation extra unreacted antibody is removed with ultrafiltration.It is not limited to 1 micron in the diameter of this microballoon, as long as size reaches
It can be converged to thermophoresis；Material is not limited to polystyrene, as long as can antibody and egg to be measured successfully be had no effect on for antibody attachment
The active material of white marker can be used, and antibody concentration and incubation temperature time are not limited to concrete numerical value described in this example,
Change with reference to actual use antibody brand batch and specific experiment condition.
The present embodiment is prepared for 11 kinds of immune microspheres using above step, to detect above-mentioned 11 kinds of markers respectively, will take
1.1 μ L of patients serum divide equally 11 parts after diluting 100 times, mix be incubated at room temperature 1 hour with 11 kinds of immune microspheres respectively, will have
There is the microballoon of the antibody and capture testing protein marker of fluorescent marker to be incubated, fluorescent marker is carried out to protein marker.
And it is detected using the detecting system of the various embodiments described above.Above-mentioned step is repeated to 10 ovarian cancer patients and 10 Healthy Peoples
Suddenly, 11 kinds of albumen marker expression amounts in 20 blood serum samples are measured, shown in Fig. 7 and Fig. 8.Due to the heterogeneity of cancer,
The marker expression of specific each patient's serum is not exactly the same, but generally expression quantity is apparently higher than healthy sample.Important
It is each protein marker as single cancer detection standard, accuracy rate is not high.Using the sum of expressing quantity in 11 as inspection
Mark is accurate, and survey can accurately distinguish oophoroma and healthy sample.It will be greatly improved using the marker for more having diagnostic significance
Accuracy rate of diagnosis, but cost increases with marker quantity and is risen, especially for the more rare costliness of antibody of certain marker.
The detection method of the present embodiment, everyone only needs 1ng antibody to each marker, and cost is not necessarily to other expensive reagents less than 1 yuan.
So far, it has been combined preferred embodiment shown in the drawings and describes technical scheme of the present invention, still, this field
Technical staff is it is easily understood that protection scope of the present invention is expressly not limited to these specific implementation modes.Without departing from this
Under the premise of the principle of invention, those skilled in the art can make the relevant technologies feature equivalent change or replacement, these
Technical solution after change or replacement is fallen within protection scope of the present invention.
Claims (10)
1. a kind of based on microsphere supported particle detection systems, which is characterized in that including heating unit, sample bin chamber unit,
In,
The heating unit, to the sample heating into the sample bin chamber unit；
The immune microsphere fluid for being combined with target biological molecules is mounted in the sample bin chamber unit, the immune microsphere is anti-
Body or aptamer are fixed on microsphere surface and are made, and the immune microsphere for being combined with target biological molecules uses fluorescent marker,
After the heating unit heats the sample bin chamber unit, thermophoretic effect is generated in the sample bin chamber unit, it will be described outer
It secretes body and converges in the lower side of temperature in the sample bin chamber unit, by the indoor target biological molecules that are combined with of sample
The fluorescence signal amplification marked on immune microsphere, to be detected.
2. according to claim 1 based on microsphere supported particle detection systems, which is characterized in that the system also includes
Signal gathering unit, the signal gathering unit obtain amplified fluorescence signal, obtain the mesh that immune microsphere surface is combined
Mark biomolecule abundance, by by various aptamers respectively from the different targets on the immune microsphere surface for being combined with target biological molecules
Protein binding obtains testing result.
3. according to claim 2 based on microsphere supported particle detection systems, which is characterized in that the sample warehouse list
Member includes loading the immune microsphere for being combined with target biological molecules and to provide the closed sample room for generating thermophoretic effect space,
The sample room includes：To close the sample room and build up the second thermal conductive surface of the micro-nano particle.
4. according to claim 3 based on microsphere supported particle detection systems, which is characterized in that second thermal conductive surface
Neighbouring temperature is less than the temperature of the micro-nano particle fluid, to be combined with target biological molecules with described in second thermal conductive surface
Immune microsphere fluid between generate the temperature difference, generate thermophoretic effect, drive and be combined with the immune microspheres of target biological molecules to the
Two thermal conductive surface displacements.
5. according to claim 3 based on microsphere supported particle detection systems, which is characterized in that the heating unit is
Laser on the outside of the sample bin chamber unit is set, is irradiated to the sample bin chamber unit, light beam passes sequentially through described
It is combined with the immune microsphere fluid and the second thermal conductive surface of target biological molecules, the target biological molecules that are combined with to be immunized
Microspheres solution generates thermophoretic effect.
6. according to claim 5 based on microsphere supported particle detection systems, which is characterized in that also wrap the sample room
It includes：To close the first thermal conductive surface of the sample room, second thermal conductive surface and the first thermal conductive surface can be such that light beam passes through.
7. according to claim 6 based on microsphere supported particle detection systems, which is characterized in that second thermal conductive surface
It is sapphire material or diamond material for transparent material；
First thermal conductive surface is any one of glass, polymethyl methacrylate, dimethyl silicone polymer, sapphire or appoints
Several combinations.
8. according to claim 1-7 any one of them based on microsphere supported particle detection systems, which is characterized in that described micro-
Ball is polystyrene microsphere.
9. a kind of based on microsphere supported particle detecting method, which is characterized in that including：
Immune microsphere is prepared, microballoon and antibody or aptamer are incubated jointly, so that antibody or aptamer is fixed on microsphere surface, is exempted from
Epidemic disease microballoon；
Immune microsphere and detected sample are incubated, on the target protein or nucleic acid in detected sample and immune microsphere
Antibody or aptamer are specifically bound, to be fixed on immune microsphere；
The immune microsphere obtained above for being combined with target biological molecules is combined with the antibody or aptamer for carrying fluorophor, it will
Target biological molecules mark fluorescent on immune microsphere；
The immune microsphere sample for being combined with target biological molecules loaded in sample bin chamber unit is heated, in sample warehouse
Unit generates thermophoretic effect, the immune microsphere for being combined with target biological molecules is converged in low in the sample bin chamber unit
Warm side, and so that signal is amplified since fluorescent marker is enriched with, to be detected.
10. according to claim 9 based on microsphere supported particle detecting method, which is characterized in that by the sample
The immune microsphere for being combined with target biological molecules that low temperature side in product warehouse unit is built up, acquisition are combined with target organism point
The immune microsphere corresponding index information of son simultaneously analyzes corresponding index.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109374890A (en) * | 2018-11-07 | 2019-02-22 | 国家纳米科学中心 | Breast cancer detection system and method based on the extracellular vesica detection of thermophoresis |
| CN110106233A (en) * | 2019-03-25 | 2019-08-09 | 杭州师范大学 | A kind of digital pcr detection method of extracellular vesica/excretion body |
| CN111999225A (en) * | 2019-12-19 | 2020-11-27 | 瑞芯智造(深圳)科技有限公司 | Method for detecting concentration of micro-nano particles |
| CN113188959A (en) * | 2021-04-29 | 2021-07-30 | 国家纳米科学中心 | Micro-nano biological particle thermophoresis detection device and method |
| CN113588938A (en) * | 2021-09-09 | 2021-11-02 | 徐州医科大学 | Immunomagnetic bead and preparation method and application thereof |
| CN113766970A (en) * | 2019-04-25 | 2021-12-07 | 公立大学法人大阪 | Method and system for collecting fine objects |
| CN115485544A (en) * | 2020-05-08 | 2022-12-16 | 公立大学法人大阪 | Detection device, detection system and detection method for tiny objects |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101428197A (en) * | 2007-10-09 | 2009-05-13 | 古河电子北美公司 | Thermophoretic fractionalization of small particles |
| CN104313139A (en) * | 2014-10-14 | 2015-01-28 | 深圳先进技术研究院 | Monomolecular cell detection method and application thereof as well as monomolecular cell detection kit |
| US20150316480A1 (en) * | 2014-05-05 | 2015-11-05 | Nanotemper Technologies Gmbh | Thermophoresis measurements in nanoliterdroplets |
| US20150322499A1 (en) * | 2014-05-12 | 2015-11-12 | National Yang-Ming University | Method for analyzing molecule using thermophoresis |
| CN208140539U (en) * | 2018-04-08 | 2018-11-23 | 国家纳米科学中心 | Based on microsphere supported particle detection systems |
-
2018
- 2018-04-08 CN CN201810306124.0A patent/CN108593910B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101428197A (en) * | 2007-10-09 | 2009-05-13 | 古河电子北美公司 | Thermophoretic fractionalization of small particles |
| US20150316480A1 (en) * | 2014-05-05 | 2015-11-05 | Nanotemper Technologies Gmbh | Thermophoresis measurements in nanoliterdroplets |
| US20150322499A1 (en) * | 2014-05-12 | 2015-11-12 | National Yang-Ming University | Method for analyzing molecule using thermophoresis |
| CN104313139A (en) * | 2014-10-14 | 2015-01-28 | 深圳先进技术研究院 | Monomolecular cell detection method and application thereof as well as monomolecular cell detection kit |
| CN208140539U (en) * | 2018-04-08 | 2018-11-23 | 国家纳米科学中心 | Based on microsphere supported particle detection systems |
Non-Patent Citations (2)
| Title |
|---|
| GA-YEON LEE ET AL.: "Thermophoretic immunoassay based on autodisplayed Z-domains for the diagnosis of C-reactive protein", 《SENSORS AND ACRUATORS B》 * |
| STEFAN DUHR ET AL.: "Two-dimensional colloidal crystals formed by thermophoresis and convection", 《APPLIED PHYSICS LETTERS》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109374890A (en) * | 2018-11-07 | 2019-02-22 | 国家纳米科学中心 | Breast cancer detection system and method based on the extracellular vesica detection of thermophoresis |
| CN110106233A (en) * | 2019-03-25 | 2019-08-09 | 杭州师范大学 | A kind of digital pcr detection method of extracellular vesica/excretion body |
| CN113766970A (en) * | 2019-04-25 | 2021-12-07 | 公立大学法人大阪 | Method and system for collecting fine objects |
| CN111999225A (en) * | 2019-12-19 | 2020-11-27 | 瑞芯智造(深圳)科技有限公司 | Method for detecting concentration of micro-nano particles |
| WO2021120944A1 (en) * | 2019-12-19 | 2021-06-24 | 瑞芯智造(深圳)科技有限公司 | Method for measuring concentration of micro/nano particles |
| CN115485544A (en) * | 2020-05-08 | 2022-12-16 | 公立大学法人大阪 | Detection device, detection system and detection method for tiny objects |
| CN113188959A (en) * | 2021-04-29 | 2021-07-30 | 国家纳米科学中心 | Micro-nano biological particle thermophoresis detection device and method |
| CN113588938A (en) * | 2021-09-09 | 2021-11-02 | 徐州医科大学 | Immunomagnetic bead and preparation method and application thereof |
| CN113588938B (en) * | 2021-09-09 | 2023-10-27 | 徐州医科大学 | Immunomagnetic bead and preparation method and application thereof |
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