CN107430052A - Particles for imaging analytical equipment - Google Patents
Particles for imaging analytical equipment Download PDFInfo
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- CN107430052A CN107430052A CN201680003512.0A CN201680003512A CN107430052A CN 107430052 A CN107430052 A CN 107430052A CN 201680003512 A CN201680003512 A CN 201680003512A CN 107430052 A CN107430052 A CN 107430052A
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/39—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using tunable lasers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
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- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
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- Spectroscopy & Molecular Physics (AREA)
- Dispersion Chemistry (AREA)
- Optics & Photonics (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention relates to a kind of particles for imaging analytical equipment, specifically, there is provided particles for imaging analytical equipment, described device are analyzed particle properties such as particle size, shapes based on the image shot by image collecting device to particle.Therefore, the present invention includes：Sample capillary, it is injected with analysis object particle；Objective table, the sample capillary is placed with thereon；Underlying light source, it is to irradiation light below the sample capillary；Optical system, using the underlying light source to sample capillary irradiation light, the particle on sample capillary that the light passes through accordingly causes light scattering, and carries out paths chosen to the image based on the scattering light by image collecting device for it；Image collecting device, shot in special time from the optical system and obtain the scattering optical image of paths chosen；And computer, it analyzes particle properties using the result of the scattering optical image frame progress signal transacting in the related input information of analysis object particle and the special time shot to described image harvester.
Description
Technical field
The present invention relates to one kind to be imaged (imaging) particle analysing device, is analyzed in particular to a kind of particles for imaging
Device, described device analyze particle based on the image shot using image collecting device (camera) to particle
The particle properties such as size, shape.
Background technology
Particle analysing device is the device for analyzing the characteristics of particle such as size, the shape of particle.The accurate survey of particle size
It is essential when being scheduled on kinetics of polymerization (generation, growth and the cohesion of particle), in addition, particle size analysis is being slapped
It is used as important analysis information in terms of the physics, chemistry, engineering properties of holding final material
In the method being measured to the particle of nanosized, with regard to dynamic light scattering (Dynamic light
Scattering, DLS) for method, by laser (laser) by light directive nano-particle, and to the degree of the light scattering
It is measured, so as to determine the size of particle.The shortcomings that methods described, is, it is necessary to be measured from all angles, not only numb
It is tired, and the light released may impact to nano-particle, so as to which the confidence level of the Nanoparticle Size determined is not high.
In addition, the problem of DLS devices, is, it is harsher to the composition of laser optical system, arrangement and the requirement of preservation,
And maintenance and repair is costly.In addition, the problem of DLS devices is, angle of scattering and measure angle are fixed, and are difficult to ensure that measure knot
The credibility of fruit, and can not also test.On DLS theoretical foundation, angle of scattering and the relatively low thing of dependence for determining angle
Although there is also exist more, thus, only matter for the definitely untrustworthy material of the measured value in specific angle
The credibility of Partical size determination result can be ensured by the value being measured in multiple angles.
In addition, for DLS devices, the actual change of the sample occurred in Partical size determination can not be observed, therefore
With restriction fundamentally when understanding phenomenon, and it is limited in, the sample just influenceed by LASER Light Source is (for example, by laser
The sample of decomposition, have vital biota sample etc.) for, it can not be analyzed.
The content of the invention
Therefore, the present invention is in order to solve described problem, and is proposed to meet the demand, and its object is to provide one
Kind particles for imaging analytical equipment, described device analyze grain based on the image shot to particle by image collecting device
The size of son, the particle properties such as shape.
The purpose of the present invention is not limited to purpose mentioned above, NM other objects of the present invention and advantage
Understood by following explanation, and practitioner can be expressly understood that according to following record, pass through embodiments of the invention
Can definitely it understand.Furthermore it is possible to be readily understood by, objects and advantages of the present invention in right by going out
Existing method and combinations thereof can be achieved.
For realizing the embodiments of the invention of the purpose, it is related to particles for imaging analytical equipment, and described device includes：
Sample capillary, it is injected with analysis object particle；Objective table, the sample capillary is placed with thereon；Underlying light source, its is right
Irradiation light below the sample capillary；Optical system, its using the underlying light source to sample capillary irradiation light, accordingly
The particle on sample capillary that the light passes through causes light scattering, and by image collecting device to based on the scattering light
Image carries out paths chosen；Image collecting device, shot in special time from the optical system and obtain dissipating for paths chosen
Penetrate optical image；And computer, it is using the related input information of the analysis object particle and to described image harvester
Scattering optical image frame (frame) in the special time of shooting carries out the result of signal transacting to analyze particle properties.
In addition, an alternative embodiment of the invention is related to particles for imaging analytical equipment, described device includes：Sample, it is wrapped
Include analysis object particle；Objective table, the sample is placed with thereon；Light source, it is arranged at the upper side of the sample or bottom
Side is so as to the sample irradiation reflected light or transmitted light；Optical system, it is utilizing shape of the light source to sample irradiation light
Under state, paths chosen is carried out to the particle image on sample by image collecting device；Image collecting device, it is to from the light
The particle image that system obtains paths chosen is shot；And computer, it shoots using to described image harvester
Particle image carry out signal transacting result to analyze particle properties, and the objective table or optical system are adjusted
So that the variable focal point of the particle, and the different focus particle images of image collecting device shooting are closed in real time
Into so as to analyze particle properties.
In addition, the still another embodiment of the present invention is characterised by, including：Sample capillary, it is injected with analysis pair
As particle；Objective table, the sample capillary is placed with thereon；LASER Light Source, it is under particles track analytical model, to described
The side irradiation light of sample capillary；Underlying light source, its under particles for imaging analytical model to the sample capillary below
Irradiation light；Optical system, its under particles track analytical model using the LASER Light Source to sample capillary irradiation light, accordingly
The particle on sample capillary that the light passes through causes light scattering, and by image collecting device to based on the scattering light
Particle image carries out paths chosen, and under particles for imaging analytical model, is irradiated using the underlying light source to sample capillary
Light, the particle on sample capillary that the light passes through accordingly cause light scattering, and by image collecting device to based on described
The particle image for scattering light carries out paths chosen；Image collecting device, it is under particles track analytical model, in special time
Shooting obtains the scattering light bead sub-image of paths chosen from the optical system, and under particles for imaging analytical model, specific
Shooting obtains the scattering optical image of paths chosen from the optical system in time；And computer, it is analyzed in particles track
Under pattern, using the related input information of the analysis object particle and in the special time of described image harvester shooting
Scattering light particle image frame carry out the result of signal transacting to follow the trail of particle movement, and particle properties is analyzed, in particles for imaging
Under analytical model, using the related input information of the analysis object particle and described image harvester is shot it is specific when
In the result of scattering optical image frame progress signal transacting analyze particle properties.
In addition, the still another embodiment of the present invention is characterised by, including：Sample, it includes analyzing object particle；Carry
Thing platform, the sample is placed with thereon；LASER Light Source, it irradiates under particles track analytical model to the side of the sample
Light；Light source, it is under real-time multifocal particle analysis pattern, to the sample irradiation reflected light or transmitted light；Optical system,
It utilizes the LASER Light Source to sample irradiation light under particles track analytical model, the grain on sample that the light passes through accordingly
Son causes light scattering, and carries out paths chosen to the particle image based on the scattering light by image collecting device, and in reality
When multifocal particle analysis pattern under, in the state of sample irradiation light, passing through image collecting device pair using the light source
Particle image on sample carries out paths chosen；Image collecting device, it is under particles track analytical model, in special time
Shooting obtains the scattering light bead sub-image of paths chosen from the optical system, and under real-time multifocal particle analysis pattern,
Shooting obtains the particle image of paths chosen from the optical system；And computer, it is under particles track analytical model, profit
With the scattering light in the related input information of analysis object particle and the special time shot to described image harvester
The result that particle image frame carries out signal transacting moves to follow the trail of particle, and analyzes particle properties, in real-time multifocal particle point
Under analysis pattern, the result that signal transacting is carried out using the particle image shot to described image harvester is special to analyze particle
Property, the objective table or optical system are adjusted under real-time multifocal particle analysis pattern, so that the focus of the particle
It is variable, and the different focus particle images of image collecting device shooting are synthesized in real time, so as to analyze particle properties.
In addition, the still another embodiment of the present invention is characterised by, including：Sample, it includes analyzing object particle；Carry
Thing platform, the sample is placed with thereon；Light source, its under particles for imaging analytical model or real-time multifocal particle analysis pattern,
Transmitted light is irradiated to the lower side of the sample, or reflected light is irradiated to the upper side of the sample；Optical system, it is being imaged
Using the light source to sample irradiation light under particle analysis pattern, the particle on sample that the light passes through accordingly causes light to dissipate
Penetrate, and paths chosen is carried out to the image based on the scattering light by image collecting device, and in real-time multifocal particle point
Under analysis pattern, using the light source in the state of sample irradiation light, by image collecting device to the particle shadow on sample
As carrying out paths chosen；Image collecting device, it is shot from the optics under particles for imaging analytical model in special time
System obtains the scattering optical image of paths chosen, and under real-time multifocal particle analysis pattern, shoots from the optical system
Obtain the particle image of paths chosen；And computer, it utilizes the analysis object particle under particles for imaging analytical model
Scattering optical image frame in related input information and the special time shot to described image harvester carries out signal transacting
Result analyze particle properties, under real-time multifocal particle analysis pattern, utilize what described image harvester was shot
The result that particle image carries out signal transacting analyzes particle properties；The load is adjusted under real-time multifocal particle analysis pattern
Thing platform or optical system, so that the variable focal point of the particle, and in real time to the different focuses of image collecting device shooting
Particle image is synthesized, so as to analyze particle properties.
In addition, the still another embodiment of the present invention is characterised by, including：Sample, it includes analyzing object particle；Carry
Thing platform, the sample is placed with thereon；LASER Light Source, it irradiates under particles track analytical model to the side of the sample
Light；Light source, it irradiates under particles for imaging analytical model or real-time multifocal particle analysis pattern to the lower side of the sample
Transmitted light, or reflected light is irradiated to the upper side of the sample；Optical system, it is under particles track analytical model using described
For LASER Light Source to sample irradiation light, the particle on sample that the light passes through accordingly causes light scattering, and passes through image collector
Put and paths chosen is carried out to the particle image based on the scattering light, and under particles for imaging analytical model, utilize the light source
To sample irradiation light, the light passes through accordingly the particle on sample causes light scattering, and by image collecting device to based on
The image of the scattering light carries out paths chosen, under real-time multifocal particle analysis pattern, is utilizing the light source to sample
In the state of irradiation light, paths chosen is carried out to the particle image on sample by image collecting device；Image collecting device, its
Under particles track analytical model, shot in special time from the optical system and obtain the scattering light particle shadow of paths chosen
Picture, under particles for imaging analytical model, shot in special time from the optical system and obtain the scattering shadow of paths chosen
Picture, under real-time multifocal particle analysis pattern, shoot from the optical system and obtain the particle image of paths chosen；And meter
Calculation machine, it is adopted using the related input information of the analysis object particle and to described image under particles track analytical model
The result that scattering light particle image frame in the special time of acquisition means shooting carries out signal transacting moves to follow the trail of particle, and point
Particle properties is analysed, under particles for imaging analytical model, using the related input information of the analysis object particle and to the figure
As the result of the scattering optical image frame progress signal transacting in special time that harvester is shot analyzes particle properties, in reality
When multifocal particle analysis pattern under, utilize the particle image that is shot to described image harvester to carry out the result of signal transacting
To analyze particle properties；The objective table or optical system are adjusted under real-time multifocal particle analysis pattern, so that institute
The variable focal point of particle is stated, and the different focus particle images of image collecting device shooting are synthesized in real time, so as to analyze
Particle properties.
The particles track analytical equipment of the present invention is imaged (imaging) mode by details in a play not acted out on stage, but told through dialogues (Dark Field) and can carried out
The nano-particle specificity analysises such as Nanoparticle Size measure, and in a liquid state can be to obtaining the nanometer of wide scope distribution
The respective particle size of particle and quantity are analyzed, and the quantity of different particle size particles is visualized, so as to
It is shown in graphical form to user, and enables a user to by real-time imaging directly to observe each particle
Distribution, dilution property, the relatively low sample of density (concentration) can be used, so as to reduce product cost and economy, according to
The image shot by image collecting device to the mobility of particle is tracked, so as to pass through display user
Visually can be visible, and the quantity of particle can be counted under low concentration environment.
In the particles track analytical equipment of the present invention, carried out relative to the specific length axis of orientation of sample with special angle
Light irradiation, thus the scope of scattering light bead sub-image needed for particle properties analysis can be extended (expansion of laser light hot spot
(Laser Spot) sample mensuration region).
For the particles for imaging analytical equipment of the present invention, while real space image is observed, reciprocal can be analyzed
Space scattering information, and the actual change of caused sample in Partical size determination can be observed.In addition, because do not make
With the laser optical system of complexity, so without the composition of laser optical system, arrangement, maintenance and repair etc., and will not be because of light
Sample damage occurs for source, and also sample to being decomposed by LASER Light Source or by other influences etc. can be carried out unconfined
Analysis.Can in addition, also being turned into and the measure at unmeasured low measure angle by existing DLS modes Partical size determination device
Energy.In addition, existing DLS modes Partical size determination device determines just getable information by multiple, by once determining just
It may obtain, and the sample that can be a few μ l to sample (sample) standard of the light microscope for routine carries out particle
Size measurement.
The present invention's is had the effect that based on real-time multifocal particles for imaging analytical equipment：In the absence of particles for imaging
(do not increase specification (spec) or not under conditions of the part change of analytical equipment, compound can carry based on real-time multifocal imaging
Particle analysis function, and the sharp parts (surface boundary) of particle can be clearly shot, and analysis object grain can be expanded
The particle properties analyst coverage such as subcategory, shape of particle can be enriched by the image collecting device filmed image based on imaging and surveyed
The market demand such as fixed, overcomes existing microscopical shorter lens focus (lens focus) scope, can analyze multiple types and
The particle properties of size, and it can be seen that the size and form of actual particle.
The set composite of the present invention has the effect that：According to analytical model common component, so as to perform its function,
And then compared to manufacture particles track analytical equipment and particles for imaging analytical equipment and real-time multifocal particle analysing device respectively, energy
Manufacturing cost is enough reduced, and in particles track analytical equipment and particles for imaging analytical equipment and real-time multifocal particle analysing device
Structure, specification, function, processing procedure need not apply any deformation can also realize, and performed respectively in set composite
When particles track analytical model and particles for imaging analytical model and real-time multifocal particle analysis pattern, in part, processing procedure
Between any interference is not present, so as to analyze particle properties in the case where not producing function reduction.
For the sample capillary installation module of the present invention, being readily able to will be smaller big with 1mm X 1mm X 80mm
Small sample capillary is equipped on analytical equipment, so as to assign user's operation ease and handling easiness.As long as will
The sample capillary for being injected with sample is positioned over sample capillary installation module, therefore enables to sample required when changing
The prior operation of preparation of samples such as cleaning it is relatively simple and easy, module is installed by sample capillary and enables to irradiation light accurate
Sample capillary really is focused on so as to assign measure accuracy and confidence level, relative to laser light irradiation axis and sample hair
Tubule side axle causes the various change of angle of laser beam irradiation desirably, as particles track analytical model and particles for imaging
The sample capillary that can be used in analytical model in the form of sharing installs module, reduces components number, so as to
Reduce manufacturing cost.
Brief description of the drawings
Fig. 1 is one embodiment pie graph for being directed to the particles track analytical equipment according to the present invention.
Fig. 2 is the stereogram for the particles track analytical equipment for showing Fig. 1.
Fig. 3 is the explanation figure for the image for obtaining and handling in the particles track analytical equipment for show Fig. 1.
Fig. 4 is the pie graph according to another embodiment of the particles track analytical equipment of the present invention.
Fig. 5 is the pie graph according to the still another embodiment of the particles track analytical equipment of the present invention.
Fig. 6 is the pie graph according to one embodiment of the particles for imaging analytical equipment of the present invention.
Fig. 7 is the stereogram for the particles for imaging analytical equipment for showing Fig. 6.
Fig. 8 is explanation figure of the displaying according to the particles for imaging analysis method of the present invention.
Fig. 9 is the real-time multifocal filming image of particles for imaging analytical equipment and the explanation figure of building-up process for showing Fig. 6.
Figure 10 is the set composite with particles track analytical model and particles for imaging analytical model according to the present invention
The stereogram of one embodiment.
Figure 11 is one embodiment stereogram according to sample capillary (capillary) the installation module of the present invention.
Figure 12 a to Figure 12 c are another embodiment stereograms according to the sample capillary installation module of the present invention.
Figure 13 is one embodiment stereogram according to the set composite with sample capillary installation module of the present invention.
Embodiment
In the following description, in order that obtaining of the invention be understood more fully hereinafter and showing the specific detailed of the present invention
Content, even if without these specific details, also it is readily able to implement the present invention according to their deformation, this leads for technology
In domain with usual knowledge personnel for be self-evident.In addition, when illustrating of the invention, for related to the present invention
Illustrating in the case of being judged as unnecessarily obscuring the purport of the present invention for known technology, it is detailed to omit its
Explanation.
Hereinafter, referring to the drawings to being described in detail according to a preferred embodiment of the invention, and to understand according to this hair
Illustrated centered on part necessary to bright operation and effect.
Fig. 1 is one embodiment pie graph for being directed to the particles track analytical equipment according to the present invention, and Fig. 2 is displaying Fig. 1
Particles track analytical equipment stereogram, Fig. 3 be in the particles track analytical equipment for be illustrated in Fig. 1 obtain and handle image
Explanation figure.
As shown in figure 1, LASER Light Source 11, objective table 12, sample hair are included according to the particles track analytical equipment of the present invention
Tubule (capillary) 13, optical system 14, image collecting device 15 and computer 16 etc..
The particles track analytical equipment of the present invention also includes microscope ocular (microscope oculars), thereby using
Person can directly use eye observation sample.
Although it has not been shown on the figures, still computer 16 also includes image capture plate (capture board), signal transacting
Portion, display (display) etc..Here, image capture plate and signal processing part may also set up in image collecting device 15.
For the particles track analytical equipment of the present invention, contain (dissolving) analysis object grain to the injection of sample capillary 13
The solution (suspension) of sub (sample), and in the state of this sample capillary 13 is positioned on objective table 12, to swash
The light (laser beam (laser beam)) of radiant 11 is irradiated by the form of sample capillary 13, and is located at this laser
The particle of the carry out Brownian movement (brownian motion) of the inner space for the sample capillary 13 that beam passes through causes light to carry out
Scattering, and paths chosen is carried out to the particle based on the scattering light using optical system 14, so as to pass through at the appointed time
Image collecting device 15 is shot the (image frame of the particle moved in the form of Brownian movement to scattering light bead sub-image
(frame) (capture) is captured, example：30~60 seconds).Also, the regulation shot by computer 16 to image collecting device 15
Scattering light particle image frame in time carries out signal transacting, so as to which the movement to each particle is tracked, and to particle
The particle properties such as size, shape, concentration, number are analyzed (measure), so as to by the display of computer 16 to user
Vision offer is provided.
The analysis object particle of the present invention can be the material (nanoparticle) of nanometer unit, and the particle is big
Small can be several nanometers (nanometer)~thousands of nanometers (example：1nm~2000nm) all categories particle.Illustratively
Property, analysis object particle can be opened as biological nano material (bio-nanoparticles) suitable for drug delivery system
Hair, viral vaccine, nanotoxicology and Biological indicators, protein cohesion, extracellular cyst (allochthon (Exosomes) and cell
Microvesicle) etc..
The particles track analytical equipment of the present invention is by light scattering (light scattering) and Brownian movement (brownian
Motion) it is applied to microscope (microscope).Detailed description below is carried out to it.
According to scattering light particle image frame, the position of the particle changed over time is followed the trail of using image tracing method, it is excellent
Selection of land tracking particle centre position, the scattering light particle image frame be by image collecting device 15 at the appointed time to
What the particle of Brownian movement form movement was shot and obtained.And difference is calculated according to the shift position of the particle of tracking
The displacement of time particle.Also, with according to the mean square displacement (MSD of different time particle displacement:Mean Squared
Displacement based on), calculated using Stokes-Einstein relation (Stocks-Einstein relation)
Diffusion coefficient (diffusion coefficient), so as to obtain Hydrodynamic diameter (hydrodynamic diameter),
So as to determine the size of particle.Also, based on the particle size of the measure, real-time rendering particle size distribution curve
(particle size distribution profile), and vision offer is carried out to user by display.
With the mean square displacement (MSD being directed to according to different time particle displacement of the present invention:Mean Squared
Displacement based on), calculated using Stokes-Einstein relation (Stocks-Einstein relation)
Diffusion coefficient (diffusion coefficient), so as to obtain Hydrodynamic diameter (hydrodynamic diameter),
So as to determine the size of particle.The algorithm (algorithm) is illustrated as follows.
Stokes-einstein formula (Stocks-Einstein relation) is such as following【Numerical expression 1】It is shown.
【Numerical expression 1】
Here, DmRepresent diffusion coefficient (diffusion co-efficient), KBRepresent Boltzmann constant
(boltzman constant), T represent temperature (temperature), and η represents viscosity (viscocity), dnRepresent that particle is big
Small (Hydrodynamic diameter, sphere equivalent hydrodynamic diameter particle size).
The random motion of particle in solution, i.e. Brownian movement are related to the mean square displacement of particle.In addition, particle is according to the time
Spatially (Gaussian distribution) spreads in solution in the form of Gaussian Profile, now, makes for each time
The value of Gaussian Profile morphological character caused by must spreading is such as following as mean square displacement【Numerical expression 2】It is shown.
【Numerical expression 2】
<MSD2>=2Dmt
Here,<MSD2>Represent mean square displacement, DmDiffusion coefficient is represented, t represents the time.
As it appears from the above, it can be obtained according to the scattering light particle image frame shot by image acquisition device 15 in solution
The mean square displacement of the particle of Brownian movement is carried out, by what is so tried to achieve【Numerical expression 2】Mean square displacement be substituting to【Numerical expression 1】Si Tuo
Ke Si-einstein formula, then it can determine the size of particle.
As it was previously stated, according to scattering light particle image frame, the particle average moving distance of two-dimensional phase is utilized【X, y】To obtain
Diffusion coefficient, in fact, particle carries out three-dimensional Brownian movement in solution.Thus, it is contemplated that clapped to by image collecting device 15
The scattering light particle image frame taken the photograph is bidimensional image, so as to such as following【Numerical expression 3】Shown applicable two-dimentional variable, so as to stoke
This-einstein formula deformed, and then can export Partical size determination formula.
【Numerical expression 3】
Here, t may also indicate that ' 1/FPS (frame per second, number of pictures per second).
LASER Light Source 11 can be implemented as laser diode (laser diode) etc., and the laser diode has 642nm
The wave band of (red), 532 (greens), 488nm (blueness), 405nm (purple) etc..
Optical system 14 can be implemented as object lens etc., and the object lens have 20 multiplying powers (x20).
Image collecting device 15 can be implemented as CCD camera (CCD camera), electron multiplication CCD (Electron
Multiplied Charged Coupled Device, EMCCD), cmos cameras (CMOS camera) etc..For example, can be with
Use (super) high speed for shooting the frame of the normal image harvester of 30 frame (30fps) per second, shooting 1,000 frame per second~3,000
Image collecting device.
Sample capillary 13 can be implemented as the capillary of lineal hexahedral shape, and be surrounded to it by outer layer of glass
Inner space injection sample, so as to make its both ends seal.The apparent size of sample capillary 13 can be illustratively
1mm X 1mm X 80mm, also, length 80mm can be according to changes such as analysis subject sample species.And sample capillary 13
As long as outer layer of glass thickness it is known in regular supplies level used in microscope.Thus, sample of the invention
Capillary 13 is 1mm X 1mm X 80mm sizes, and it is then 0.08cm to be scaled volume3, as 0.08ml, even if injection
Having that the inner space volume of sample is identical with apparent volume, maximum can carry out particle properties analysis using 0.08ml samples,
Only user is required using less sample size.In addition, the sample capillary 13 of the present invention is used as capillary pipe structure, with hair
Tubule phenomenon sucks to sample, so as to easily carry out sample injection, and because is 1mm X 1mm X 80mm
Smaller size, so in sample capillary particle movement do not influenceed by convection phenomena, grain is enabled to by Brownian movement
Son is mobile, thus allows for the more accurately particle properties such as Partical size determination analysis.
Objective table 12 is connected with motor (motor) (being not shown in accompanying drawing), and drives control by the motor of computer 16
System or user's directly operation etc. can move up and down (the variable focal point of example, optical system 14 and image collecting device 15
When), move left and right, the movement of six direction of principal axis such as side pitching (tilting) (example, grasping during horizontal tilt horizontal).
In addition, as in Fig. 2 stereograms, the specific part of the objective table 12 of sample capillary 13 is being placed formed with groove
(hole), it is inscape extra in particles track analytical equipment, shared with particles for imaging analytical model to realize
Property, the particles for imaging analytical model is that reference picture 10 illustrates later.In other words, in particles track analytical equipment
In, because the irradiation light of Laser Power Devices 11 passes through the side of sample capillary 13, performing Figure 10 particles for imaging analysis
During pattern, it is necessary to which so that the irradiation light of underlying light source 102 is from the bottom of sample capillary 105, portion side passes through on the side, therefore excellent
Selection of land, groove is formed in objective table 12.
The advantages of particles track analytical equipment of the foregoing present invention, is as follows.
The nanoparticles such as Nanoparticle Size measure can be carried out by being imaged (imaging) mode by details in a play not acted out on stage, but told through dialogues (Dark Field)
Sub-feature is analyzed, and in a liquid state can be to obtaining the respective particle size of nano-particle and quantity of wide scope distribution
Analyzed, and the quantity of different particle size particles is visualized, so as to be shown with chart to user, and
And enable a user to directly observe the distribution of each particle by real-time imaging, dilution property, density can be used
(concentration) relatively low sample, so as to reduce product cost and economy, according to passing through movement of the image collecting device to particle
Property the image that is shot be tracked, so as to visually can be visible by display user, and low dense
The quantity of particle can be counted under degree environment.
Next, another main feature again of the present invention is illustrated.
As shown in drawings, sample capillary 13 is positioned on objective table 12, so that the laser beam of LASER Light Source 11
The side of (light) to the length direction of sample capillary 13 is irradiated, in the present invention, relative to the light source of LASER Light Source 11
The side axle of axle and sample capillary 13, laser beam is irradiated with predetermined angular (crooked trunnion axis).In other words, using light source to
During sample irradiation light, light irradiation is carried out at a particular angle relative to the specific length axis of orientation of sample.
In other words, although it is also possible that the side axle of the light source axle of LASER Light Source 11 and sample capillary 13 is with 90 °
Angle along level carry out light irradiation, but it is described in the case of, pass through the sample of the smaller size with 1mm X 1mm X 80mm
The visual field (field of view, FOV) that the light of product capillary 13 to scatter light bead sub-image diminishes, so as to want from image
The scattering light bead sub-image that harvester 15 obtains may diminish.
Thus, in the present invention, in order to expand the region of the scattering light bead sub-image required for particle properties analysis, sentence is changed
Talk about, for expansion of laser light hot spot (Laser Spot) sample mensuration region, relative to the specific length axis of orientation of sample with spy
Determine angle and realize light irradiation.
Relative to the specific length axis of orientation of sample, illumination is realized with special angle (example, preferably 10 °~80 °, 20 °)
Penetrate, the laser by the side of the length direction of the sample capillary 13 on this mode objective table relative to LASER Light Source 11
Beam direction of illumination is crooked, or can use following manner：So that the laser beam direction of illumination of LASER Light Source 11 is relative to sample
The side surface direction of capillary 13 is crooked.
Side axle relative to the light source axle and sample capillary 13 of the LASER Light Source 11 of the present invention is (askew with defined angle
Oblique trunnion axis) irradiation laser beam, thus relative to the sample capillary 13 with 1mm X 1mm X 80mm sizes, optical system
The 14 and FOV (field of view) of image collecting device 15, i.e. laser facula (Laser Spot) sample mensuration region are illustrated
Property it is minimum can be 30 μm of 50 μm of X, 80 μm of X etc..
The scattering light bead sub-image that Fig. 3 a displayings are shot by image collecting device 15, in other words, show relative to sharp
The side axle of the light source axle of radiant 11 and sample capillary 13 irradiates laser beam so as to obtain with predetermined angular (crooked trunnion axis)
The scattering light bead sub-image obtained.In fig. 3 a, particle is shown in the diagonal of image collecting device filmed image, and it is right
The angle of linea angulata refers to illumination firing angle.
Next, as shown in Figure 3 b, in the present invention, the differentiation algorithm that each particle is tracked is prompted.Fig. 3 b exhibitions
Show that the scattering light bead sub-image shot to image collecting device 15 is applicable the different particles differentiation tracing algorithm of the present invention and handled
Image, such image provided by the display of computer 16 to user.
In Fig. 3 b image, line represents the motion track caused by the Brownian movement of different particles, and in image, left side line
Represent the motion track of the first particle, in image, right-hand line represents the motion track of the second particle.Can by such image
So that user is intuitive to see different particle behavior analysis results.
For the different particles of the present invention distinguish tracing algorithm, matched (Pattern matching) by image mode
Particle label (labelling) is assigned, for example, assigning identification information to particle, so as to be made a distinction to each particle, and then is led
Go out the motion track of each particle.
As the example of different particles differentiation tracing algorithm, on the scattering light particle image frame obtained at the appointed time,
By with changing over time and particle that adjacent image interframe closely moves change is characterized as identical particle, and to described
The particle of discrimination assigns intrinsic indivedual identification informations, and the position of the particle of same identification information is endowed to each image frame
Put coordinate to be drawn (plotting), so as to generate track.
Different particles distinguish another example of Trajectory Arithmetic, and the scattering light particle obtained at the appointed time influences frame
On, it will change over time and there is adjacent image interframe the particle of identical image to be characterized as identical particle, and to described
The particle of discrimination assigns intrinsic indivedual identification informations, and the position of the particle of same identification information is endowed to each image frame
Put coordinate to be drawn (plotting), so as to generate track.
Next, in the present invention, during to sample 13 irradiation light of capillary, multiple LASER Light Sources can be used.As showing
Example, laser beam synthesizer is set between two LASER Light Sources and the side of sample capillary 13, so that from each laser
The laser beam that light source is irradiated is inputted to laser beam synthesizer, and can be to cause the laser beam synthesized in laser beam synthesizer
The form exported to the side of sample capillary 13 carries out light irradiation.As another example, set causing two LASER Light Sources
It is placed in the state of the side of sample capillary 13, to pass through sample capillary from the laser beam that each LASER Light Source is irradiated
The form of the same area of the inner space of pipe 13 is focused (focusing), thus allows for light irradiation.
As described above, it is to improve the light that sample capillary 13 obtains using multiple LASER Light Sources in the present invention.
For example, during using 30mW laser, bright reduction that sample capillary 13 obtains, so that can be obtained by image collecting device 15
The scattering light bead sub-image obtained has the limitation of maximum 33 frame per second.Thus, in the present invention, carried using multiple LASER Light Sources
The light that high sample capillary 13 obtains, and so that the frame of the scattering light bead sub-image obtained by image collecting device 15
Increase to more than 33 frame per second, so as to shorten the overall minute required for particles track analysis.
Next, the particles track analysis method of the present invention is illustrated.
The phase of the solution (suspension) for the analysis object particle (sample) that (dissolving) injects to sample capillary 13 will be included
Close information and condition determination relevant parameter (parameter) input computer 16.Here, sample message and condition determination parameter can
To be temperature, number of pictures per second, analyze amount of images, image collecting device configuration information, image intensity (intensity), sample
Information (example, water, ethanol (alcohol), toluene (toluene) etc.) etc..Preferably, according to the sample message so inputted and
Condition determination parameter causes light irradiation control, filming image control, image frame signal transacting to control and differently set, so as to
It can be handled.
Also, as it was previously stated, to be positioned on objective table 12 sample capillary 13, relative to the light source of LASER Light Source 11
The side axle of axle and sample capillary 13 irradiates laser beam with defined angle (crooked trunnion axis).Thus, passed through positioned at laser beam
The particle of the carry out Brownian movement (brownian motion) of the inner space for the sample capillary 13 crossed causes light scattering.
Afterwards, as it was previously stated, paths chosen is carried out by optical system 14 to the particle based on scattering light, so as to provide
(the shadow of the particle moved in the form of Brownian movement is shot to scattering light bead sub-image by image collecting device 15 in time
As frame (frame) acquisition (capture), example：30~60 seconds).
Also, the scattering light particle image frame in the stipulated time shot by computer 16 to image collecting device 15 enters
Row signal transacting, so as to which the movement to each particle is tracked, and the particle such as the size to particle, shape, concentration, number is special
Property analyzed (measure), so as to carry out vision offer to user by the display of computer 16.Now, as it was previously stated,
With according to the mean square displacement (MSD of different time particle displacement:Mean Squared Displacement) based on, make
Diffusion coefficient (diffusion is calculated with Stokes-einstein formula (Stocks-Einstein relation)
Coefficient), so as to obtain Hydrodynamic diameter (hydrodynamic diameter), so as to determine the size of particle.
Next, reference picture 4, is illustrated to another embodiment of the particles track analytical equipment according to the present invention.
Fig. 4 is another embodiment pie graph according to the particles track analytical equipment of the present invention.
As shown in figure 4, particles track analytical equipment according to another embodiment of the invention is said including reference picture 1
Bright LASER Light Source 11, objective table 12, sample capillary 13, optical system 14, image collecting device 15, computer 16, and
Scattering light filter house (filtering) 17 is set between optical system 14 and image collecting device 15.
Scattering light is produced due to irradiating the laser beam of LASER Light Source 11 to sample capillary 13, now, passes through sample hair
The countless particles of tubule 13 and can produce multiple scattering interference.For so a variety of scattering interference, to pass through image
Based on the scattering light bead sub-image that harvester 15 obtains when analyzing particle properties, cause to hinder accuracy, credible
Degree, the influence of reappearance.
As shown in drawings, in the present invention, the configuration scattering light filtering between optical system 14 and image collecting device 15
During portion 17 so that scattering light filter house 17 focuses on place (also referred to as imaging plane) positioned at the output of optical system 14, and causes
Required location of the image collecting device 15 positioned at the rear end of scattering light filter house 17.
Scattering light selects caused by scattering the particle for the sample capillary 13 that light filter house 17 to input from optical system 14
Selecting property by other words so that by multiple scattering disturb scattering light be cut off, so as to prevent multiple scattering interference make
Into influence.Scattering light filter house 17 can be implemented as pin hole (pin hole), and the pin hole has several nanometers to several microns big
It is small.Optical system 14 can be implemented as object lens.
Next, reference picture 5, has another embodiment to say the particles track analytical equipment according to the present invention
It is bright.
Fig. 5 is the pie graph according to the still another embodiment of the particles track analytical equipment of the present invention.
As shown in figure 5, carried out according to the particles track analytical equipment of the still another embodiment of the present invention including reference picture 1
LASER Light Source 11, objective table 12, sample capillary 13, optical system 14, image collecting device 15, the computer 16 of explanation, and
And including light scattering preventing portion 18 and level meter 19.
For the particles track analysis of the present invention, in the state of sample capillary 13 is positioned on objective table 12,
It is irradiated in the form of causing the light (laser beam) of LASER Light Source 11 by sample capillary 13, now, passes through sample capillary
The light of pipe 13 is reflected in internal part of exterior object or particles track analytical equipment etc., and sample is incident upon so as to reenter
Capillary 13.Because reflected light is re-shoot in such sample capillary 13, so that the scattering light particle based on reflected light
Image mixes with the scattering light bead sub-image of the light based on LASER Light Source 11, so as to as so that the degree of accuracy in particle properties measure
The reason for decline.
Thus, in the present invention, on objective table 12, light is set to reflect preventing portion in the rear end place of sample capillary 13
18, preventing portion 18 is reflected by light by the light (laser beam) of sample capillary 13 and absorbed.Such light reflection preventing portion 18 can
It is embodied as with material, material, any part of characteristic absorbed to light (laser beam), for example, it may be attachment black
Plate (sheet) of cloth etc..
In addition, as illustrated, the present invention particles track analytical equipment objective table 12 on set level meter 19, so as to
The horizontality of objective table 12 is grasped, and when the horizontality of objective table 12 is crooked, the axle of objective table 12 is adjusted, from
And horizontality can be corrected.For example, when realizing level meter 19 with electronic type, the level of level meter 19 is grasped by computer 16
State, motor is automatically controlled so as to be adjusted to the axle of objective table 12, or during mechanically realizing level meter 19, user
The horizontality of level meter 19 is grasped, motor can be directly controlled by computer 16 so as to be adjusted to the axle of objective table 12.
In the present invention as described above, the horizontality of objective table 12 is corrected by level meter 19, so as to anti-
The sample capillary 13 being only positioned on objective table 12 tilts to side, and can prevent the sample capillary in non-standard state
Convection phenomena occurs for the inside of pipe 13.As a result, the focusing of the light (laser beam) irradiated from light source 11 to sample capillary 13 is suitable
Profit is realized, and prevents convection phenomena, so as to improve particle properties measure performance.
Next, reference picture 6 illustrates to Fig. 8 to the particles for imaging analytical equipment of the present invention.
Fig. 6 is according to the pie graph of one embodiment of the particles for imaging analytical equipment of the present invention, and Fig. 7 is displaying Fig. 6
The stereogram of particles for imaging analytical equipment, Fig. 8 are explanation figure of the displaying according to the particles for imaging analytical equipment of the present invention, and Fig. 9 is
Show Fig. 6 real-time multifocal filming image of particles for imaging analytical equipment and the explanation figure of building-up process.
As shown in fig. 6, underlying light source 61, objective table 62, sample hair are included according to the particles for imaging analytical equipment of the present invention
Tubule (capillary) 63, optical system 64, image collecting device 65 and computer 66 etc..In addition, particles for imaging analytical equipment
It may also include top power supply 67.
The particles for imaging analytical equipment of the present invention also includes microscope ocular (microscope oculars), thereby using
Person also can directly use eye observation sample.
Although it has not been shown on the figures, still computer 66 also includes image capture plate (capture board), signal transacting
Portion, display (display) etc..Here, image capture plate and signal processing part may also set up in image collecting device 65.
For the particles for imaging analytical equipment of the present invention, contain (dissolving) analysis object grain to the injection of sample capillary 63
The solution (suspension) of sub (sample), and in the state of this sample capillary 63 is positioned on objective table 62, under causing
The light of portion's light source 61 is irradiated through the form of the sample capillary 63, and is caused positioned at the inside of sample capillary 63
The particle of the carry out Brownian movement (brownian motion) in space causes transmitted light to be scattered, and the scattering light is led to
Cross optical system 64 and carry out paths chosen, so as to be shot at the appointed time by image collecting device 65 to scattering light
(scattering optical image frame (frame) captures (capture), example：30~60 seconds).Also, by computer 66 to image collector
Put scattering optical image frame in the stipulated time of 65 shootings and carry out signal transacting, so as to the size of particle, shape, concentration, individual
The particle properties such as number are analyzed (measure), so as to carry out vision offer to user by the display of computer 66.
The analysis object particle of the present invention can be the material (nanoparticle) of nanometer unit, and the particle is big
Small can be several nanometers (nanometer)~thousands of nanometers (example：1nm~2000nm) all categories particle.Illustratively
Property, analysis object particle can be opened as biological nano material (bio-nanoparticles) suitable for drug delivery system
Hair, viral vaccine, nanotoxicology and Biological indicators, protein cohesion, extracellular cyst (allochthon (Exosomes) and cell
Microvesicle) etc..
Underlying light source 61 can be implemented as 300W Halogen lamp LED (halogen lamp) etc. as general white light source.
Top light source 67 is embodied as general white light source, or can be implemented as optical diode (laser diode) etc..Institute
State top light source 67 to use when for that can not realize that the sample of mobile measure carries out particle properties analysis by transmitted light, example
Such as, it can be separated caused by the temperature change of metal solid solution analysis, solder (Solder) melting point analysis, abnormity
Phase separation analysis of macromolecule mixture or block copolymerization polymeric membrane (film) etc..In other words, metal and height are being contained
Top light source 67 is used during the particle properties analysis of impermeable (impermeableness) material of molecule.
Optical system 64 can be implemented as object lens etc., and the object lens have 20 multiplying powers (x20).
Image collecting device 65 can be implemented as CCD camera (CCD camera), electron multiplication CCD (Electron
Multiplied Charged Coupled Device, EMCCD), CMOS camera (CMOS camera) etc..For example, can be with
Use (super) high speed for shooting the frame of the normal image harvester of 30 frame (30fps) per second, shooting 1,000 frame per second~3,000
Image collecting device.
Sample capillary 63 can be implemented as the capillary of lineal hexahedral shape, and be surrounded to it by outer layer of glass
Inner space injection sample, so as to make its both ends seal.The apparent size of sample capillary 63 can be illustratively
1mm X 1mm X 80mm, also, length 80mm can be according to changes such as analysis subject sample species.And sample capillary 63
As long as outer layer of glass thickness it is known in regular supplies level used in microscope.Thus, sample of the invention
Capillary 63 is 1mm X 1mm X 80mm sizes, and it is then 0.08cm to be scaled volume3, as 0.08ml, even if injection
Having that the inner space volume of sample is identical with apparent volume, maximum can carry out particle properties analysis using 0.08ml samples,
Only user is required using less sample size.In addition, the sample capillary 63 of the present invention is used as capillary pipe structure, with hair
Tubule phenomenon sucks to sample, so as to easily carry out sample injection, and because is 1mm X 1mm X 80mm
Smaller size, so particle in sample capillary is moved not by convection phenomena image, grain is enabled to by Brownian movement
Son motion, thus allow for the more accurately particle properties such as Partical size determination analysis.
Objective table 62 is connected with motor (motor) (being not shown in accompanying drawing), and drives control by the motor of computer 66
System or user's directly operation etc. can move up and down (the variable focal point of example, optical system 64 and image collecting device 65
When), move left and right, the movement of six direction of principal axis such as side pitching (tilting) (example, grasping during horizontal tilt horizontal).
The particles for imaging molecular device of the present invention is stated light scattering (light scattering) and Brownian movement
(brownian motion) is applied to microscope (microscope).Detailed description below is carried out to it.
It is irradiated in the form of causing the light of underlying light source 61 to be transmitted to sample capillary 63, so as to be located at sample capillary
The particle of the progress Brownian movement of the inner space of pipe 63 causes transmission light-scattering layer.
Thus, obtained by image collecting device 65 and carry out the generation of change over time that the particle of Brownian movement is triggered
The change of scattering light, so as to carry out Fourier (Fourier) analysis to the scattering optical image frame, and then to scatter light
The formal layout that time/spatial information is decomposed, and by determining the size of nano-particle to its parsing.
In other words, the difference image between the scattering optical image frame obtained by image collecting device 65 is tried to achieve
(difference image), (example：When being shot 2 seconds with per second 3,000 frames, difference image 5,999), to the difference diagram
As carrying out two-dimensional Fourier transform.The scattered light intensity of each pixel (Pixel) unit can be perceived by such difference image
The change of degree, the spatial distribution of scattering light can be obtained by the two-dimensional Fourier transform of difference image, to scattering light space point
Cloth is parsed, so as to obtain the space time information of the scattering light according to time change.
Hereafter, to Fourier transformation result perform azimuth scan (Azimuthal Scan), the azimuth scan try to achieve from
The center of circle in center being averaged apart from residing value to identical pixel.Fig. 8 left side (a) is displaying two-dimensional Fourier transform value
The concept map of azimuth scan, Fig. 8 right side (b) are to show over time to change and the absolute value changes of caused azimuth scan value
Chart.In Fig. 8 left side (a), from the central q of Fourier transformation value1、q2、q3It is corresponding with angle of scattering respectively.Thus, at this
In invention, the Fourier transformation value tried to achieve according to difference image includes polygonal scattered information, is become Fourier according to this concentric circles
The average value for the aggregate value that value adds up is changed, is drawn, is then obtained such as Fig. 8 right side (b) in graphical form according to time change
As a result.
Particle size is determined using numeric value analysis skill and technique to the azimuth scan value thus tried to achieve, now, use is following【Numerical expression
4】。
【Numerical expression 4】
Here, | FD(q, Δ t) |2Represent in reciprocal space according to the absolute of the azimuth scan value of the distance q at a distance of center
Value, characteristics of the A (q) on particle, B (q) represent image collecting device caused noise (noise) in itself, and τ (q) represents relaxation
Time (relaxtion time).
【Numerical expression 4】Relaxation time τ (q) relative to diffusion coefficient Dm(diffusionco-efficient) and reciprocal is empty
Between positional value (q) have it is following【Numerical expression 5】Relation.
【Numerical expression 5】
【Numerical expression 5】Diffusion coefficient DmWith with it is described【Numerical expression 1】Stokes-einstein formula (Stokes-
Einstein relation) identical relation.
With described【Numerical expression 4】、【Numerical expression 5】And【Numerical expression 1】Based on, it is right【Numerical expression 4】Perform non-linear curve fitting
(nonlinear curve fitting), so as to speculate【Numerical expression 4】Three kinds of constants (unknown number), that is, speculate A (q), B (q), τ
(q), and by the τ (q) of supposition value substitute into【Numerical expression 5】, so as to basis【Numerical expression 1】Speculate particle size dh。
Furthermore it is preferred that when carrying out the particles for imaging analysis of the present invention, sample message and condition determination parameter are inputted
To computer 66.In other words, the solution for the analysis object particle (sample) that (dissolving) injects to sample capillary 63 will be contained
(suspension) relevant information and condition determination relevant parameter input computer 66.Here, sample message and condition determination parameter can
To be image number, average time, image section (image interval), radius section (radius interval), temperature
It is worth input, light source displacements sensor, the removal of background dust image etc..Preferably, according to the sample message and survey thus inputted
Fixed condition parameter causes light irradiation control, filming image control, image frame signal transacting to control and differently set, so as to
Access processing.
The particles for imaging analytical equipment of the present invention as described above has the following advantages that.
While real space image is observed, reciprocal space scattered information can be analyzed, and particle size can be surveyed
The actual change of caused sample is observed in fixed.In addition, because without using complicated laser optical system, without swashing
The composition of light optical system, arrangement, maintenance and repair etc., and sample damage will not occur because of light source, also can be to passing through laser
Light source is decomposed or sample by other influences etc. carries out unconfined analysis.It is in addition, big by existing DLS modes particle
It is small measure device and it is unmeasured it is low measure angle measure be also possibly realized.In addition, existing DLS modes Partical size determination dress
Put by the repeatedly getable information of measure, may be obtained by once determining, and can be to the optics for routine
The sample that microscopical sample (sample) standard is a few μ l carries out Partical size determination.
Next, another principal character again of the present invention is illustrated.
Fig. 9 is the real-time multifocal filming image of particles for imaging analytical equipment and the explanation figure of building-up process for showing Fig. 6.
As it appears from the above, the particles for imaging device application light scattering and Brownian movement of the present invention, big so as to not only carry particle
The particle properties analytic functions such as small measure, and carry based on real-time multifocal particles for imaging analytic function.
As shown in figure 9, motor control etc. carried out by using computer 66 so that objective table 62 is to optical system 64
Direction hoisting depth, so that real-time focal point (focusing on (focusing)) is variable, and for being positioned on objective table 62
The particle of sample 91, not confocal particle image is shot in the form of real time high-speed by image collecting device 65.And lead to
The not confocal particle image of the composograph harvester 65 of computer 66 shooting is crossed, so as to divide the characteristic of particle
Analysis.
In other words, in the real-time multifocal particle analysis pattern of the present invention so that variable focal point, so that each
The sharp parts of focus and particle (that is, surface boundary, edge (edge)) match, now, real time high-speed it is each to focusing on
The not confocal particle image of individual particle surface boundary is shot.Also, to be shot not by image collecting device 65
With being synthesized centered on each particle surface boundary of focus particle image, so as to manufacture single particle image, and then analyze
Its particle properties.
The particle properties can be the size of particle, area, major axis, short axle, sphericity, girth, form etc..In addition, point
Analysis object particle can be a few nm units particle the solid powder of several μm~a few mm scopes or the streaming of several μm of units it is thin
Born of the same parents etc., and foreign bodies detection of metal dust and plate (panel) etc. can be applied to.
As real-time focal point as described above (focusing) flexible method, the motor control that can be carried out by using computer 66
System etc. so that objective table 62 is to the direction of optical system 64 according to the rules apart from lifting or falling head, and be used as another
Example, the motor control etc. that can be carried out by using computer 66 so that optical system 64 to the direction of objective table 62 according to
Predetermined distance is lifted or falling head.Certainly, user also can manually adjust objective table 62 or optical system 64, from
And enable to real-time focal point (focusing) variable.
Under the real-time multifocal particle analysis pattern of the present invention, image collecting device 65 for shoot 1,000 frame per second with
On high-speed image sampling device.
In addition, according to species for analyzing object particle etc., the light source that sample is irradiated can alternatively be used and be based on
The transmitted light of underlying light source 61 or the reflected light based on top light source 67.
The present invention as described above based on real-time multifocal particles for imaging analytical equipment the advantages of it is as follows.
(rule are not increased under conditions of the part change of the particles for imaging analytical equipment illustrated in the absence of reference picture 6
Lattice (spec)), compound it can carry based on real-time multifocal particles for imaging analytic function, and can clearly shoot particle
Sharp parts (surface boundary), and the analysis particle properties analyst coverage such as object particle kind can be expanded, by based on into
The image collecting device filmed image of picture can the market demand such as sufficient shape of particle measure, overcome existing microscopical shorter
Lens focus (lens focus) scope, can analyze multiple types and the particle properties of size, and it can be seen that actual particle
Size and form.
In addition, this hair that particles for imaging analytical equipment of the invention or reference picture 9 that reference picture 6 illustrates illustrate
The scattering light filter house that bright implementation multifocal particle analysing device enables to reference picture 4 to illustrate is arranged at optical system
Between 64 and image collecting device 65.Omit and such scattering light filter house is illustrated.
In addition, this hair that particles for imaging analytical equipment of the invention or reference picture 9 that reference picture 6 illustrates illustrate
The level meter that bright real-time multifocal particle analysing device enables to reference picture 5 to illustrate is arranged on objective table 62.Save
Slightly such level meter is illustrated.
Figure 10 is the set composite with particles track analytical model and particles for imaging analytical model according to the present invention
The stereogram of one embodiment.
As shown in Figure 10, according to the compound dress with particles track analytical model and particles for imaging analytical model of the present invention
Put including LASER Light Source 101, underlying light source 102, top light source 103, objective table 104, sample capillary 105, optical system
106th, image collecting device 107 and it is not shown in computer of accompanying drawing etc..
In addition, Figure 10 set composite can also include the scattering light filter house that illustrates of reference picture 4, reference picture 5 is entered
The light reflection preventing portion and level meter of row explanation.
The set composite of the present invention causes the function and ginseng referring to figs. 1 to Fig. 5 particles track analytical equipments illustrated
Performed in one apparatus according to the function of Fig. 6 to Fig. 8 particles for imaging analytical equipments illustrated, compound execution particle chases after
Track analytical model and particles for imaging analytical model.
Certainly, set composite of the invention is able to carry out the work(referring to figs. 1 to the 5 particles track analytical equipments illustrated
The real-time multifocal particle analysis function that can be illustrated with reference picture 9.
In addition, the set composite of the present invention is able to carry out referring to figs. 1 to Fig. 5 particles track analytical equipments illustrated
The real-time multifocal that function, the reference picture 9 for the particles for imaging analytical equipment that function, reference picture 6 to Fig. 8 illustrate illustrate
The function of particle analysing device.
In other words, set composite of the invention performs particles track analytical model and particles for imaging analytical model, is holding
During row particles track analytical model, LASER Light Source 101, objective table 104, sample capillary 105, optical system 106, image are used
Harvester 107, when performing particles for imaging analytical model, using underlying light source 102, objective table 104, sample capillary 105,
Optical system 106, image collecting device 107.
As described above, the set composite of the present invention is according to analytical model common component, so as to perform its function, and then
Compared to particles track analytical equipment and particles for imaging analytical equipment and real-time multifocal particle analysing device is manufactured respectively, can drop
Low manufacturing cost, and in particles track analytical equipment and particles for imaging analytical equipment and the knot of real-time multifocal particle analysing device
Structure, specification, function, processing procedure, which need not apply any deformation, can also realize, and perform particle respectively in set composite
When follow-up analysis pattern and particles for imaging analytical model and real-time multifocal particle analysis pattern, between part, processing procedure not
Any interference be present, so as to analyze particle properties in the case where not producing function reduction.
Next, the sample capillary installation module to the present invention is specifically described.
Figure 11 is to be according to one embodiment stereogram of the sample capillary installation module of the present invention, Figure 12 a to Figure 12 c
According to another embodiment stereogram of the sample capillary of present invention installation module, Figure 13 is that have sample according to the present invention
One embodiment stereogram of the set composite of capillary installation module.
The particles track illustrated referring to figs. 1 to Fig. 5 is all equipped on according to the sample capillary of present invention installation module
What particles for imaging analytical equipment that analytical equipment, reference picture 6 to Fig. 9 illustrate, reference picture 10 illustrated chases after with particle
On the objective table 12,62,104 of the set composite of track analytical model and particles for imaging analytical model so that sample capillary 13,
63rd, 105 it is positioned over the sample capillary installation module.
As shown in figure 11, according to the sample capillary of the first embodiment of the present invention install module 200 upper side along
For its length direction formed with mounting groove 201, sample capillary 13 is installed on the mounting groove 201, and mould is installed from sample capillary
The lateral side opposite side of side one of block 200 is formed with through slot 202, so that installing module 200 towards sample capillary
Side irradiation light it is incident, and projected by sample capillary 13.
As figure 12 a shows, according to the second embodiment of the present invention sample capillary installation module 300 upper side along
For its length direction formed with mounting groove 301, sample capillary 13 is installed on the mounting groove 301, and mould is installed from sample capillary
The lateral side opposite side of side one of block 300 formed with through slot 302, and as sample capillary installation module 300 side
The blocked construction of opposite side, reflect preventing portion 303 formed with the light that reference picture 5 illustrates in this side opposite side.Change sentence
Talk about, as figure 12 a shows, when the side opposite side of sample capillary installation module 300 forms light reflection preventing portion 303, i.e.,
Make particles track analytical equipment be not provided with light reflection preventing portion 18 can also.
As shown in Figure 12b, according to the third embodiment of the invention sample capillary installation module 400 upper side along
Its length direction is installed on the mounting groove 401 formed with mounting groove 401, sample capillary 13,63,105, from sample capillary
The lateral side opposite side of side one of module 400 is installed formed with through slot 402, installed in sample capillary under module 400
Portion face is formed with the lower channel 403 with particular size.
In other words, the load of lower channel 403 and Fig. 7 of sample capillary installation module 400 particles for imaging analytical equipment
The hole of the objective table 104 of the hole of thing platform 62 or Figure 10 set composite matches, and the light irradiated from underlying light source 61,102 passes through
The hole of objective table 104, the lower channel 403 of sample capillary installation module 400, through sample capillary 63,105.Such figure
What 12b sample capillary installation module 400 can share in particles track analytical model and particles for imaging analytical model
Form is used.
As figure 12 a shows, Figure 12 b sample capillary installation module 400 is the side that sample capillary installs module 400
The blocked construction of opposite side, this side opposite side can reflect preventing portion formed with light.
As shown in fig. 12 c, according to the fourth embodiment of the invention sample capillary installation module 500 upper side along
Its length direction is installed on the mounting groove 501 formed with mounting groove 501, sample capillary 13,63,105, from sample capillary
The lateral side opposite side of side one of module 500 is installed formed with through slot 502, in the upper of sample capillary installation module 500
Portion side is formed with lid 503, and the lid 503 is formed with the upper slot 504 with particular size.
In other words, Figure 12 c sample capillary installation module 500 is in order that its internal sample capillary must be arranged at
13rd, 63,105 outside not by the foreign matters such as dust permeate and lid 503 is set, and cause come from sample capillary 13,63,
105 scattering light projects to upper direction, therefore lid 503 is formed with the upper slot 504 as scattering light exit path.
In addition, the lid 503 of Figure 12 c sample capillary installation module 500 fixtured specimen in a manner of hinge (hinge)
The side and the side of lid 503 of capillary installation module 500, so as to as the construction for make it that lid 503 is opened and closed.Example
Such as, in the state of lid 503 is opened so that sample capillary 13,63,105 rises and can start to survey after closing lid 503
It is fixed.
In addition, the sample capillary as Figure 12 c installs the structure that the both ends of the mounting groove 501 of module 500 are blocked, energy
Enough cut through the both ends of mounting groove 501 possibly into the foreign matter such as dust permeate.
In addition, the bottom surfaces in Figure 12 c sample capillary installation module 500 can be also formed with particular size
Portion's groove, the structure that the side opposite side as sample capillary installation module 500 is blocked, can be in the side opposite side
Form light reflection preventing portion.
In addition, Figure 11 to Figure 12 c sample capillary installs module 200,300,400,500 to prevent light from reflecting, energy
Enough carry out being similar to Al2O3It is surface-treated etc. unglazed anodic oxidation (Anodizing).
In addition, as shown in figure 13, Figure 11 to Figure 12 c sample capillary installs module 200,300,400,500 by tight
Gu part is fixed and is equipped on objective table 12,62,104.
As another example, in fig. 13, sample capillary installs the bottom appearance of module 200,300,400,500
Alongst can also formed with tongue and groove, the upper side of objective table 12,62,104 also can formed with sample hair
The corresponding tongue and groove of tongue and groove of tubule installation module 200,300,400,500.Described sample capillary is enabled to pacify
Die-filling piece 200,300,400,500 of tongue and groove is fastened and fixed with the tongue and groove of objective table 12,62,104.By such recessed
Tongue realizes limiter (stopper) and is oriented to fastening function.
In addition, as shown in figure 13, for the light irradiation direction of fixed laser light source 11,101, fixed component can be equipped on
901.Described LASER Light Source fixed component 901, which may be disposed at, to carry out illumination to the side of sample capillary 13,63,105
The arbitrary position penetrated, for example, the upper side of objective table 12,62,104 can be arranged at etc., the sample capillary 13,63,
105 are equipped on sample capillary installation module 200,300,400,500.
For the present invention, module 200,300,400,500 and fixed component 901 are installed using sample capillary, relatively
Laser beam is irradiated with defined angle (crooked trunnion axis) in laser light irradiation axis and sample capillary side axle, so as to assign
Give device form convenience and accuracy, the sample capillary 13,63,105 along sample capillary install module 200,
300th, 400,500 length direction carries, and the fixed component 901 is consolidated to the light irradiation direction of LASER Light Source 11,101
It is fixed.In other words, when wanting to be set as any value by the angle of laser light irradiation axis and sample capillary side axle, with
The installation direction of the crooked LASER Light Source fixed component 901 of form to match with this angle, or the installation of crooked sample capillary
The installation direction of module 200,300,400,500.
The advantages of above-mentioned sample capillary of the invention installs module is as follows.
It is readily able to the sample capillary with the smaller sizes of 1mm X 1mm X 80mm being equipped on analytical equipment, so as to
User's operation ease and handling easiness can be assigned.As long as the sample capillary for being injected with sample is positioned over sample hair
Tubule install module, therefore enable to the prior operation of the preparation of samples such as sample cleaning required when changing relatively simple and
Simplicity, module is installed by sample capillary and enables to irradiation light to focus on sample capillary exactly so as to assign survey
Determine accuracy and confidence level, relative to laser light irradiation axis and sample capillary side axle laser beam is irradiated desirably
The various change of angle, it can be used as in particles track analytical model and particles for imaging analytical model in the form of sharing
Sample capillary installation module, reduce components number, so as to reduce manufacturing cost.
In addition, in method of the invention as previously shown, computer programing can be passed through.And realize described program
Code (code) and code segment (code segment) can be easy by the computer programmer (programmer) of art
Deduce on ground.In addition, the program storage of the establishment is counted in the recording medium (information recording medium) that can be readable by a computer
The interpretation of calculation machine simultaneously performs, thus come realize the present invention method.And the recording medium includes can be by computer interpretation
The recording medium of whole forms.
In addition, in detailed description of the invention, although having carried out related description to specific embodiment, do not taking off
From in the limit of the scope of the present invention, various deformation can be carried out naturally.Therefore, the scope of the present invention can not be by illustrated reality
Apply example limitation and limit, should be not only according to right described later, and should be according to impartial with this right
To determine.
Claims (8)
1. a kind of particles for imaging analytical equipment, it includes：
Sample capillary, it is injected with analysis object particle；
Objective table, the sample capillary is placed with thereon；
Underlying light source, it is to irradiation light below the sample capillary；
Optical system, its sample capillary using the underlying light source to sample capillary irradiation light, accordingly the light transmission
On particle cause light scattering, and paths chosen is carried out to the image based on the scattering light by image collecting device；
Image collecting device, shot in special time from the optical system and obtain the scattering optical image of paths chosen；And
Computer, it is using the related input information of the analysis object particle and to the specific of described image harvester shooting
Scattering optical image frame in time carries out the result of signal transacting to analyze particle properties.
2. particles for imaging analytical equipment according to claim 1, it is characterised in that
The computer carries out Fourier transformation to the difference image of the scattering optical image frame in special time, so as to scatter light
Change is obtained in the form of time/spatial information, time/space scattering optical information and the analysis object using the acquisition
Particle associated temperature and viscosity determine particle size.
3. particles for imaging analytical equipment according to claim 2, it is characterised in that
The computer is pushed away by the azimuth scan value of the time according to acquired in the Fourier transformation/space scattering optical information
The relaxation time of survey, reciprocal space scatter the relational expression measure particle size between optical position value and diffusion coefficient.
4. particles for imaging analytical equipment according to claim 3, it is characterised in that
The computer is by the related temperature of the diffusion coefficient tried to achieve from the relational expression, the analysis object particle and viscosity generation
Enter Stokes-einstein formula, so as to determine particle size.
5. particles for imaging analytical equipment according to claim 1, it is characterised in that
The related input information of the analysis object particle includes image number, average time, image section, radius section, temperature
Degree, sample message.
6. particles for imaging analytical equipment according to claim 1, it is characterised in that
Also including scattering light filter house, the scattering light filter house is arranged between the optical system and image collecting device,
So that the scattering optical image from optical system input selectively passes through.
7. particles for imaging analytical equipment according to claim 6, it is characterised in that
The output that the scattering light filter house is arranged at optical system focuses on place, and it is described make it that image collecting device is arranged at
Scatter the locality of the rear end of light filter house.
A kind of 8. particles for imaging analytical equipment, it is characterised in that including：
Sample, it includes analyzing object particle；
Objective table, the sample is placed with thereon；
Light source, it is arranged at the upper side of the sample or lower side, so as to the sample irradiation reflected light or transmitted light；
Optical system, its using the light source in the state of sample irradiation light, by image collecting device on sample
Particle image carries out paths chosen；
Image collecting device, it shoots to the particle image that paths chosen is obtained from the optical system；And
Computer, it analyzes particle using the result of the particle image progress signal transacting shot to described image harvester
Characteristic,
The objective table or optical system are adjusted, so that the variable focal point of the particle, and image is adopted in real time
The different focus particle images of acquisition means shooting are synthesized, so as to analyze particle properties.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160009385A KR101766838B1 (en) | 2016-01-26 | 2016-01-26 | Particle Analyzer Microscope |
| KR10-2016-0009385 | 2016-01-26 | ||
| PCT/KR2016/011439 WO2017131316A1 (en) | 2016-01-26 | 2016-10-12 | Imaging particle analyzing apparatus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107430052A true CN107430052A (en) | 2017-12-01 |
| CN107430052B CN107430052B (en) | 2020-03-27 |
Family
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201680003512.0A Expired - Fee Related CN107430052B (en) | 2016-01-26 | 2016-10-12 | Imaging particle analysis device |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR101766838B1 (en) |
| CN (1) | CN107430052B (en) |
| WO (1) | WO2017131316A1 (en) |
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| KR101858533B1 (en) | 2016-10-17 | 2018-05-16 | 윈포시스(주) | Particle Analyzer Microscope |
| KR101986903B1 (en) * | 2017-09-29 | 2019-06-07 | 포항공과대학교 산학협력단 | Appratus and methods for optical detection and analysis of particle |
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Also Published As
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| KR101766838B1 (en) | 2017-08-23 |
| WO2017131316A1 (en) | 2017-08-03 |
| CN107430052B (en) | 2020-03-27 |
| KR20170089234A (en) | 2017-08-03 |
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