CN105784646B - It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference - Google Patents
It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference Download PDFInfo
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- CN105784646B CN105784646B CN201610191384.9A CN201610191384A CN105784646B CN 105784646 B CN105784646 B CN 105784646B CN 201610191384 A CN201610191384 A CN 201610191384A CN 105784646 B CN105784646 B CN 105784646B
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- 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
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
- G01N21/554—Attenuated total reflection and using surface plasmons detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
Abstract
The invention discloses a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference.The present invention is for there is complex biological environment existing for macromolecular mass, utilize the surface plasma body resonant vibration coupling effect of noble metal nano particles pair, a kind of method is disclosed, macromolecular ambient noise is matched and rejected while this method can realize the noble metal nano particles of orientation random distribution to best shooting condition.This method may be implemented in each noble metal nano particles pair for orientation random distribution of developing while loss in the biotic environment of macromolecular mass noise jamming without target.
Description
Technical field
The invention belongs to the single molecular recognitions in life science and nano-probe technical field, are related to metal nano
The detection of probe pair is related specifically to sparse in tested surface be scattered and multiple nano metal probes that orientation is random are to most preferably enhancing
Acquisition methods while amplitude image.
Background technology
Noble metal nano particles with surface plasmon resonance effect, such as gold nanoparticle have high brightness, nothing
The features such as photobleaching, bio-compatibility are good, thus as a kind of marker for the probe biomolecule being concerned.Not only such as
This, they have unique surface plasmon resonance effect, extremely strong absorption are shown in visible near infrared band, especially
When two particles are close together, and surface plasma body resonant vibration coupling occurs, the electricity of enhancing is generated in the gap location of the particle pair
, than the delustring intensity increase several times of single particle.Therefore the use of noble metal nano particles pair is in enhancing molecular image figure
Picture and bio-sensing sensitivity etc. have important value and wide application space.But this humidification is strong
Dependent on shooting condition, it is desirable that the polarization state of exciting light, which needs to match with the axle center of the line of centres composition to particle, can just obtain
Best enhancing effect is obtained, such as shown in Figure 1, when the polarization state of exciting light is parallel with the axis of particle pair, enhancing is maximum；When
When the two is vertical, enhancing effect is minimum.Multiple particles pair, the axis of each pair of particle can be existed simultaneously in the test of sample to be tested
Space arrangement and position distribution be all random, if only being matched partial target therein, and by its image enhancement
It displays, other target images for not obtaining enhancing can be submerged and lose, it is therefore desirable to the particle of all random alignments
To all carrying out enhancing condition coupling.In addition to this, it is excellent in their sizes in the biological sample also there are large biological molecule group
Gesture is enough to flood the signal of noble metal nano particles pair.Therefore it is lost in the biotic environment for thering is macromolecular mass to interfere without target
Noble metal nano particles are detected to being the key that carry out quantitative biochemical analysis, proposition of the invention there can be macromolecular mass interference
Biotic environment in realize multi-target particle to the Auto-matching of shooting condition and without particle to the best enhancing image of loss
Develop simultaneously.
Invention content
The technical problem to be solved by the present invention is to tested surface inner orientation is realized in the biotic environment for having macromolecular mass to interfere
The automatic shooting condition matching of the best enhancing image of multiple nano metal probes pair of random alignment and simultaneously acquisition methods.
The technical scheme is that：
It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference, it is special
Sign is that noble metal nano particles are the metal nanoparticle with surface plasmon resonance effect, is continuously changed in tested plane
Become the angle between the axis of noble metal nano particles pair and incident light excitation polarization state so that expensive per a pair in tested plane
The axis of metallic obtains equal polarization state matching machine meeting in measurement accuracy.Specifically comprise the following steps：
Step 1：Set surface plasma body resonant vibration coupled wave of the wavelength of incident light as noble metal nano particles to generation
It is long；
Step 2：Projecting direction of the polarization state of incident light in tested plane is P, continuously rotates P's with the step-length of setting
The integral multiple of direction half cycle or half cycle；A step-length is often rotated, measures and records a width amplitude image；After completing all rotations,
Obtain n width amplitude images；
Step 3：The n width image that step 2 obtains about tested plane coordinate system is resetted and is overlapped；For in tested plane
All amplitude images are carried out comparison in situ by same location point, if change rate of strength is less than the threshold value of setting, the test point
Impurity is judged as, and is rejected；
Step 4：Step 3 is repeated, impurity is carried out to each point being tested in plane and judges and rejects；
Step 5：All amplitude images after impurity will be eliminated and carry out superposition in situ, the image generated after superposition is each pair of
Noble metal nano particles all obtain incident light polarization state and the matched enhancing image of its axis；
The setting principle of the step-length is：Value range is 0 to 45 degree, and step-length is set smaller, the identification essence of target
Degree is higher, and it is longer to measure the spent time；
The change rate of strength of the amplitude image for maximum value of the same measured point in n width images divided by
Minimum value；The value range of the change rate of strength threshold value of amplitude image is more than or equal to 2.
The effect of the present invention be can be highly sensitive in having complex biological environment existing for macromolecular realization by noble metal
Nano-particle develops to the multi-targets recognition of label and simultaneously.
Description of the drawings
Attached drawing 1 be between particle coupled electric field amplitudes with incident ray polarization direction and change of the particle to the angle of the line of centres
Change relationship simulation result figure.In figure：Diameter is respectively the grain that the centre of sphere spacing that two gold particles of 60nm are formed is 65nm
Son is right；X-axis is for incident ray polarization direction with particle to the angle (unit of the line of centres：Degree)；Y-axis is the sub- coupled electric field of double grains
Amplitude relative to X be 90 degree when normalization.
Attached drawing 2 is that single noble metal nano particles, large biological molecule group, noble metal are entering the test sample coexisted
Enhance change schematic diagram during ray polarised light different directions angle.
In figure：1 single noble metal nano particles；2 noble metals pair；
3 and 4：Large biological molecule group；5 are present in enhancing electric field of the particle to gap location；
Represent the polarization state direction of incident light.
Specific embodiment
Attached drawing 2 for single noble metal nano particles, large biological molecule, noble metal to the test sample coexisted in incidence
Enhance change schematic diagram during light polarization direction difference, the polarization state of incident light from the X-axis angle with tested plane coordinate system be 0 degree
It is starting point according to being rotated clockwise, 4 rotation positions is illustrated in figure.By taking a pair of of gold goal as an example, the diameter of two gold goals
Respectively 60nm, centre of sphere spacing are 65nm, as shown in Figure 1, the changing with incident ray polarization direction during rotation
Become, strong variation occurs for the surface plasma body resonant vibration coupling enhancing intensity of the sub- gap location of double grains.When polarization state and double grains
When the line of centres is parallel, coupling enhancing is maximum；And when polarization state is perpendicular to double grains subcenter line, coupling enhancing disappears.It is single
A metal nanoparticle and large biological molecule group do not have surface plasma body resonant vibration and couple enhancement effect, in rotary course,
The change rate of its oscillator intensity is much smaller than the change rate of the oscillator intensity of bimetal nano particles.It is a kind of not by the more of background interference
Extracting method while the enhancing image of target noble metal nano particles pair, it is characterized in that noble metal nano particles are with surface
The metal nanoparticle of plasma resonance effect continuously changes the axis of noble metal nano particles pair with entering in tested plane
Penetrate the angle between light excitation polarization state so that the axis per a pair of of noble metal in tested plane obtains in measurement accuracy
Polarization state matching machine meeting on an equal basis is obtained, is included the following steps：
Step 1：Set surface plasma body resonant vibration coupled wave of the wavelength of incident light as noble metal nano particles to generation
It is long；
Step 2：Projecting direction of the polarization state of incident light in tested plane is P, continuously rotates P's with the step-length of setting
The integral multiple of direction half cycle or half cycle；A step-length is often rotated, measures and records a width amplitude image；After completing all rotations,
Obtain n width amplitude images；
Step 3：The n width image that step 2 obtains about tested plane coordinate system is resetted and is overlapped；For in tested plane
All amplitude images are carried out comparison in situ by same location point, if change rate of strength is less than the threshold value of setting, the test point
Impurity is judged as, and is rejected；
Step 4：Step 3 is repeated, impurity is carried out to each point being tested in plane and judges and rejects；
Step 5：All amplitude images after impurity will be eliminated and carry out superposition in situ, the image generated after superposition is each pair of
Noble metal nano particles all obtain incident light polarization state and the matched enhancing image of its axis；
It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference, it is special
Sign is that the setting principle of step-length is：Value range is 0 to 45 degree, and step-length is set smaller, and the accuracy of identification of target is higher, is measured
The spent time is longer；
It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference, it is special
The change rate of strength for being amplitude image is maximum value divided by minimum value of the same measured point in n width images；
It is a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference, it is special
The value range for being the change rate of strength threshold value of amplitude image is more than or equal to 2.
Claims (3)
- It is 1. a kind of not by extracting method while the enhancing image of the multiple target noble metal nano particles pair of background interference, feature It is noble metal nano particles for the metal nanoparticle with surface plasmon resonance effect, continuously changes in tested plane Angle between axis and incident light the excitation polarization state of noble metal nano particles pair so that your every a pair of gold in tested plane The axis for belonging to particle obtains equal polarization state matching machine meeting in measurement accuracy, includes the following steps：Step 1：Set surface plasma body resonant vibration coupled wavelength of the wavelength of incident light as noble metal nano particles to generation；Step 2：Projecting direction of the polarization state of incident light in tested plane is P, and the direction of P is continuously rotated with the step-length of setting The integral multiple of half cycle or half cycle；A step-length is often rotated, measures and records a width amplitude image；After completing all rotations, n is obtained Width amplitude image；Step 3：The n width image that step 2 obtains about tested plane coordinate system is resetted and is overlapped；For same in tested plane All amplitude images are carried out comparison in situ by a location point, if change rate of strength is less than the threshold value of setting, which is judged to It is set to impurity, and is rejected；The change rate of strength of amplitude image for maximum value of the same measured point in n width images divided by Minimum value；Step 4：Step 3 is repeated, impurity is carried out to each point being tested in plane and judges and rejects；Step 5：All amplitude images after impurity will be eliminated and carry out superposition in situ, the image generated after superposition is your each pair of gold Belong to nano-particle and all obtain incident light polarization state and the matched enhancing image of its axis.
- It is 2. as described in claim 1 a kind of not by the same of the enhancing image of the multiple target noble metal nano particles pair of background interference When extracting method, it is characterized in that the setting principle of step-length is：Value range is 0 to 45 degree.
- It is 3. as claimed in claim 1 or 2 a kind of not by the enhancing image of the multiple target noble metal nano particles pair of background interference While extracting method, spy be the change rate of strength threshold value of amplitude image value range be more than or equal to 2.
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| CN107101942B (en) * | 2017-05-17 | 2019-04-23 | 大连理工大学 | A kind of probe being used to polarize micro-imaging based on bimetal nano particles |
| CN108414423A (en) * | 2018-04-27 | 2018-08-17 | 成都理工大学 | The scanning electron microscope imaging method of the micro-nano hole of rock |
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| US7760421B2 (en) * | 2004-04-06 | 2010-07-20 | Solaris Nanosciences, Inc. | Method and apparatus for enhancing plasmon polariton and phonon polariton resonance |
| CN102539400A (en) * | 2011-12-31 | 2012-07-04 | 广东工业大学 | High-accuracy fluorescence anisotropy microscopic imaging device and method |
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| CN104020084A (en) * | 2014-06-17 | 2014-09-03 | 大连理工大学 | Method for recognizing precious metal nano particles from dielectric medium scattering background |
| CN104484894A (en) * | 2014-11-18 | 2015-04-01 | 中国科学院大学 | Multi-wavelength lamination imaging technology facing to three-dimensional information recovery |
| CN104711314A (en) * | 2015-02-05 | 2015-06-17 | 李乐 | Method for cell imaging by adopting polarized light microscope to observe nano particles |
| WO2015134847A1 (en) * | 2014-03-07 | 2015-09-11 | Trustees Of Boston University | Polarization enhanced interferometric imaging |
| CN105182556A (en) * | 2015-09-02 | 2015-12-23 | 鲁东大学 | Multi-focus array light spot generation device and method |
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- 2016-03-28 CN CN201610191384.9A patent/CN105784646B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7760421B2 (en) * | 2004-04-06 | 2010-07-20 | Solaris Nanosciences, Inc. | Method and apparatus for enhancing plasmon polariton and phonon polariton resonance |
| CN102539400A (en) * | 2011-12-31 | 2012-07-04 | 广东工业大学 | High-accuracy fluorescence anisotropy microscopic imaging device and method |
| WO2015134847A1 (en) * | 2014-03-07 | 2015-09-11 | Trustees Of Boston University | Polarization enhanced interferometric imaging |
| CN104020085A (en) * | 2014-06-17 | 2014-09-03 | 大连理工大学 | Optical detection and microimaging method of micro-nano particles not subjected to influence of background |
| CN104020084A (en) * | 2014-06-17 | 2014-09-03 | 大连理工大学 | Method for recognizing precious metal nano particles from dielectric medium scattering background |
| CN104484894A (en) * | 2014-11-18 | 2015-04-01 | 中国科学院大学 | Multi-wavelength lamination imaging technology facing to three-dimensional information recovery |
| CN104711314A (en) * | 2015-02-05 | 2015-06-17 | 李乐 | Method for cell imaging by adopting polarized light microscope to observe nano particles |
| CN105182556A (en) * | 2015-09-02 | 2015-12-23 | 鲁东大学 | Multi-focus array light spot generation device and method |
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