CN110779894B - System, method and device for acquiring direction of plasma nanorod - Google Patents
System, method and device for acquiring direction of plasma nanorod Download PDFInfo
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- CN110779894B CN110779894B CN201911103432.4A CN201911103432A CN110779894B CN 110779894 B CN110779894 B CN 110779894B CN 201911103432 A CN201911103432 A CN 201911103432A CN 110779894 B CN110779894 B CN 110779894B
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- G—PHYSICS
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- 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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- 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
- G01N2021/4704—Angular selective
- G01N2021/4711—Multiangle measurement
- G01N2021/4714—Continuous plural angles
Abstract
The application provides a system, a method and a device for acquiring the direction of a plasma nanorod, wherein the system comprises: the device comprises an illumination light source, a first polaroid, a second polaroid, a dark-field microscope and a processing device, wherein the illumination light source and an objective lens of the dark-field microscope are oppositely arranged, an objective table of the dark-field microscope carrying nanorods is arranged between the illumination light source and the objective lens, the first polaroid is arranged between the illumination light source and the objective table, the second polaroid is arranged between the objective table and the objective lens, and the polarization direction of the first polaroid is vertical to that of the second polaroid; the processing device is used for acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles, and determining the direction of each nanorod based on the plurality of dark field images. The method and the device utilize the dark field microscope to image the nanorods at different horizontal angles under polarized illumination, can accurately estimate the direction of each nanorod, and have important significance for analyzing complex nanorod samples.
Description
Technical Field
The application relates to the field of spectral analysis, in particular to a system, a method and a device for acquiring the direction of a plasma nanorod.
Background
Plasma is a state of matter existing in parallel with solid, liquid and gaseous states, with equal numbers of positively and negatively charged particles. Plasma nanorods (e.g., gold nanorods) have excellent anisotropic optical and chemical properties, which can provide anisotropic information about the orientation of the probe and its environment, in addition to non-blinking, non-bleaching absorption and scattering properties, and are widely used as biological and biomedical imaging probes.
Dark Field Microscopy (DFM) techniques use oblique illumination to block direct light transmitted through a detail of a sample, based on the physical properties of the light, and observe the sample as reflected and diffracted light. The plasma nanorods have strong optical properties such as Local Surface Plasmon Resonance (LSPR) absorption and scattering, and have strong interaction with electromagnetic radiation, so that scattered light imaging can be obtained by the DFM technology. However, only one light spot formed by scattered light can be observed in a dark field image obtained by imaging, and currently, the scattering-based DFM imaging technology cannot provide the orientation information of the nanorods.
Disclosure of Invention
An object of the embodiments of the present application is to provide a system, a method and a device for obtaining the direction of plasma nanorods, which can accurately estimate the direction of each nanorod, thereby solving the above-mentioned technical problems.
In order to achieve the above purpose, the embodiments of the present application provide the following technical solutions:
in a first aspect, an embodiment of the present application provides a directional acquisition system for plasma nanorods, including: the device comprises an illumination light source, a first polaroid, a second polaroid, a dark-field microscope and a processing device, wherein the illumination light source is opposite to an objective lens of the dark-field microscope, a stage of the dark-field microscope carrying nanorods is arranged between the illumination light source and the objective lens, the first polaroid is arranged between the illumination light source and the stage, the second polaroid is arranged between the stage and the objective lens, and the polarization direction of the first polaroid is perpendicular to that of the second polaroid; the illumination light source is used for providing an initial illumination light beam; the processing device is used for acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles, and determining the direction of each nanorod based on the plurality of dark field images.
According to the scheme, the dark field microscope is used for imaging the nanorods at different horizontal angles under polarized illumination, so that the direction of each nanorod can be accurately estimated, and the method has important significance for analyzing a complex nanorod sample.
Optionally, the nanorods are carried on a slide, and the slide is carried on a stage of a dark-field microscope; the stage rotates the slide at preset angular increments.
Optionally, the system further includes: the first polaroid and the second polaroid are fixed on the adjusting device; the adjusting device is used for controlling the first polaroid and the second polaroid to synchronously rotate according to preset angle increment.
The horizontal angle of the nanorods refers to an angle formed between the nanorods and a reference axis, which is a horizontal axis constructed on the plane of the slide and parallel to the polarization direction of the first polarizer on the horizontal plane. In the two schemes, the first polarizing film and the second polarizing film can be synchronously rotated, so that the polarization direction of the first polarizing film on the horizontal plane is changed, the horizontal angle of the nanorod is changed, and the slide glass can be rotated through the objective table, so that the nanorod is rotated by an angle, and the horizontal angle of the nanorod is changed.
Optionally, the processing device is specifically configured to: acquiring a plurality of dark field images of nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot regions formed by the corresponding nanorods; and determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods.
Optionally, the processing device is specifically configured to: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the variation of the light intensity value with the horizontal angle of the nanorod, and the function is a cosine function.
Optionally, the processing device is specifically configured to: and generating a reconstructed image for representing the position and the direction of each nanorod by taking the direction of the nanorod at the corresponding position in the dark-field image as the long axis direction, taking the first preset length as the long axis length of the nanorod and taking the second preset length as the short axis length of the nanorod.
Optionally, the processing device is specifically configured to: converting the dark field image into a gray level image, and converting the gray level image into a binary image through threshold segmentation, wherein under the condition that the light intensity value of each pixel point in the dark field image is inconsistent, the binary image is obtained by processing the gray level image through a fuzzy c-means algorithm to obtain a corrected image with uniform illumination and then performing threshold segmentation on the corrected image; and clustering each pixel point in the binary image through a region growing algorithm to obtain a plurality of light spot regions.
Optionally, the processing device is specifically configured to: acquiring target images of nanorods acquired by a dark-field microscope at different moments, wherein the target images are images generated in the dark-field microscope after a catalyst is added into a glass slide bearing the nanorods and the nanorods generate a photocatalytic reaction based on the catalyst; and determining the change information of the nano rod in the photocatalytic reaction process according to the plurality of target images at different moments.
In a second aspect, an embodiment of the present application provides a direction-finding method for plasma nanorods, which is applied to a processing device in the direction-finding system as described in the first aspect, and the method includes: acquiring a plurality of dark field images of nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot regions formed by the corresponding nanorods; and determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods.
Under the action of linearly polarized light, the intensities of light spots formed by the same nanorod in different directions are different. According to the scheme, the dark field microscope is used for calculating the intensity value of the scattering light of the nanorods, the orientation of each nanorod is estimated according to the change of the scattering light intensity, and the direction information of the nanorods can be conveniently obtained in a time-saving manner.
Optionally, the determining the direction of each nanorod in the dark-field image according to the light intensity values of the spot areas at the same position in the multiple dark-field images and the mapping relationship between the light intensity values and the horizontal angles of the nanorods includes: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the variation of the light intensity value with the horizontal angle of the nanorod, and the function is a cosine function.
The mapping relation between the light intensity value and the horizontal angle of the nano rod can be represented by cosine function fitting, and the horizontal angle corresponding to the nano rod can be determined according to the cosine function, so that the direction of the nano rod can be obtained.
Optionally, after determining the direction of each nanorod in the dark-field image, the method further comprises: and generating a reconstructed image for representing the position and the direction of each nanorod by taking the direction of the nanorod at the corresponding position in the dark-field image as the long axis direction, taking the first preset length as the long axis length of the nanorod and taking the second preset length as the short axis length of the nanorod.
After the reconstructed image is generated, the positions and the directions of all the nanorods can be directly observed from the reconstructed image, so that the nanorods can be analyzed in the same direction accurately in the subsequent experimental study of the nanorods.
Optionally, after obtaining a plurality of dark-field images of the nanorods acquired by the dark-field microscope at different horizontal angles, the method further includes: converting the dark field image into a gray level image, and converting the gray level image into a binary image through threshold segmentation, wherein under the condition that the light intensity value of each pixel point in the dark field image is inconsistent, the binary image is obtained by processing the gray level image through a fuzzy c-means algorithm to obtain a corrected image with uniform illumination and then performing threshold segmentation on the corrected image; and clustering each pixel point in the binary image through a region growing algorithm to obtain a plurality of light spot regions.
When the illumination is not uniform, a more accurate binary image can be obtained through correction processing, so that each light spot area can be accurately obtained.
Optionally, after determining the direction of each nanorod in the dark-field image, the method further comprises: acquiring target images of nanorods acquired by a dark-field microscope at different moments, wherein the target images are images generated in the dark-field microscope after a catalyst is added into a glass slide bearing the nanorods and the nanorods generate a photocatalytic reaction based on the catalyst; and determining the change information of the nano rod in the photocatalytic reaction process according to the plurality of target images at different moments.
The nanorods generate Local Surface Plasmon Resonance (LSPR) under the action of polarized light emitted by the first polarizer and the catalyst, and photochemical conversion of the nanorod surfaces is promoted. After the direction of each nanorod is obtained, statistics can be performed according to the change information of the nanorods in different directions respectively, so that the accuracy and the stability of an analysis result are improved.
Optionally, the light intensity value of each pixel point in the speckle region is calculated by a formula
Calculating to obtain the light intensity value of the light spot region, wherein the light intensity value of the light spot region is determined by the mean value of the light intensity values of all pixel points in the region or the maximum light intensity value in the region; r, G, B respectively represent the RGB values of the corresponding pixels.
In a third aspect, an embodiment of the present application provides a directional fetching device for plasma nanorods, the device including: the image acquisition module is used for acquiring a plurality of dark field images of the nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot areas formed by the corresponding nanorods; and the direction determining module is used for determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods.
In a fourth aspect, embodiments of the present application provide a storage medium having a computer program stored thereon, where the computer program is executed by a processor to perform the method according to the second aspect.
In a fifth aspect, an embodiment of the present application provides a processing apparatus, including: a processor, a memory and a bus, the memory storing machine-readable instructions executable by the processor, the processor and the memory communicating over the bus when the processing device is running, the machine-readable instructions when executed by the processor performing the method of the second aspect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments of the present application will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and that those skilled in the art can also obtain other related drawings based on the drawings without inventive efforts.
FIG. 1 is a schematic diagram of a directional acquisition system for plasma nanorods provided in an embodiment of the present application;
FIG. 2 is a flowchart of a method for obtaining direction of plasma nanorods according to an embodiment of the present application;
FIG. 3 is a schematic view of the horizontal angle of the nanorods in the example of the present application;
FIG. 4 is another flow chart of a method for obtaining direction of plasma nanorods according to an embodiment of the present application;
FIG. 5 is another flow chart of a method for obtaining direction of plasma nanorods according to an embodiment of the present application;
fig. 6 is a schematic view of a directional acquisition apparatus for plasma nanorods according to an embodiment of the present application.
Icon: 101-an illumination source; 102-a first polarizer; 103-a second polarizer; 104-dark field microscope; 105-a processing device; 106-nanorods; 107-slide.
Detailed Description
The technical solutions in the embodiments of the present application will be described below with reference to the drawings in the embodiments of the present application.
An embodiment of the present application provides a system for obtaining a direction of a plasma nanorod, referring to fig. 1, the system includes: the device comprises an illumination light source 101, a first polaroid 102, a second polaroid 103, a dark-field microscope 104 and a processing device 105, wherein the illumination light source 101 is opposite to an objective lens of the dark-field microscope 104, a stage (not shown) of the dark-field microscope carrying nanorods 106 is arranged between the illumination light source 101 and the objective lens, the first polaroid 102 is arranged between the illumination light source 101 and the stage, the second polaroid 103 is arranged between the stage and the objective lens, and the polarization direction of the first polaroid 102 is perpendicular to the polarization direction of the second polaroid 103.
In a specific implementation, in the direction acquisition system, the illumination light source 101 is used to provide an initial illumination beam; the first polarizer 102 is configured to adjust the initial illumination light beam to obtain a first polarized light, where the first polarized light is a linearly polarized light and is incident on the surface of the nanorod 106 supported on the stage; the second polarizer 103 is used for adjusting the scattered light generated by the nanorods 106 under the action of the linearly polarized light to obtain second polarized light; the dark-field microscope 104 is used for imaging based on the second polarized light emitted by the second polarizer to obtain a dark-field image of the nanorods. The horizontal angle of the nano rod rotates according to the preset angle increment, and the dark field microscope can obtain a plurality of dark field images of the nano rod under different horizontal angles. The processing device 105 is used for acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope 104 at different horizontal angles, and determining the direction of each nanorod based on the plurality of dark field images.
It should be noted that the illumination light source may be a part of the dark field microscope, or may be a separate light source; the processing device can obtain a dark-field image from the dark-field microscope in a data copying mode or perform image transmission with the dark-field microscope through the communication interface; the polarization directions of the first polarizing plate and the second polarizing plate are not limited to be perpendicular in a strict sense, and a slight deviation may be allowed.
Alternatively, the nanorods 106 are carried on a slide 107, and the slide 107 is carried on the stage of a dark field microscope. The slide comprises a slide glass and a cover glass, the nanorods are placed in a reaction area formed by the slide glass and the cover glass, and the slide glass is placed on the object stage.
The obtaining modes of the multiple dark field images of the nano rod under different horizontal angles at least comprise the following two modes:
A. the stage of the dark field microscope controls the rotation of the slide in preset angular increments.
In the mode A, the object stage can be rotated manually, so that the nano rods can form images at a plurality of angles, the object stage can also be accurately controlled to rotate through another control device, and therefore the glass slide rotates according to the preset angle increment, and the nano rods are driven to rotate.
B. The adjustment means controls the first polarizer 102 and the second polarizer 103 to rotate synchronously at preset angular increments.
In the mode B, the direction acquiring system further includes: the first polaroid and the second polaroid are fixed on the adjusting device; the first polarizing plate and the second polarizing plate are rotated in synchronization based on the control of the adjusting means.
In one embodiment, the horizontal angle of the nanorods is changed from 0 ° to 360 ° in a preset angular increment (e.g., 5 °) on the reference axis (horizontal axis constructed on the plane of the slide parallel to the polarization direction of the first polarizer), so that dark-field images of one nanorod at the current horizontal angle can be obtained every 5 °, for a total of 72 dark-field images.
The embodiment of the application provides a direction acquisition method of a plasma nanorod based on the direction acquisition system. This example describes the method by taking gold nanorods (AuNRs) as an example, but it should be understood that the method is also applicable to nanorods formed by other metals, such as copper, zinc, silver, aluminum, etc., and thus the type of the nanorods should not be construed as limiting the examples of the present application. Referring to fig. 2, for an example of a processing device in a direction acquisition system, the method includes the following steps:
s201: and acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles. Before step S201, the method may include the steps of:
step (1): and preparing a CTAB coating AuNRs sample by adopting a seed growth method.
As an embodiment of this example, preparation of AuNRs samples in the experiment was performed in advance. 11.9ml of cetyltrimethylammonium bromide (CTAB) with a concentration of 0.2mol/L and 5ml of chloroauric acid (HAuCl) with a concentration of 2mmol/L4·4H2O) was gently mixed, and then 10. mu.L, 0.15ml, and 0.01ml of silver nitrate (AgNO) were sequentially added to the mixture3) And 0.16ml, 0.1ml of Ascorbic Acid (AA). After the solution became colorless, 0.11ml of a seed solution of gold nanoparticles was added to the solution, and then the tube was rapidly shaken for 10 seconds and left to stand for 24 hours, during which time the red color of the solution gradually changed. The reaction mixture was washed 3 times with deionized water to remove excess reagents.
Step (2): and (3) constructing a self-made flow cell by taking a glass slide and a cover glass as reaction areas.
A simple self-made flow cell was constructed based on the prepared AuNRs sample and the scattered light of the AuNRs sample was observed. In this example, 10. mu.L of AuNRs colloid (i.e., the AuNRs sample obtained in the above step (1), the 10. mu.L of AuNRs colloid including a plurality of gold nanorods) was deposited in the reaction area, the AuNRs colloid was diluted with 100-fold dilution water, after reacting for 30 minutes, the flow cell was washed with purified water, and then nitrogen gas (N-N) was used2) The flow cell is dried.
It should be understood that the above-mentioned dosage, concentration, shaking time, standing time, etc. are only specific examples, and may be appropriately adjusted according to actual conditions when applied, and the embodiment of the present application is not particularly limited thereto. In practical cases, if the prepared nanorods are already available, the preparation steps can be not separately performed.
And (3): one region containing the AuNRs sample was selected and placed under a dark field microscope for DFM imaging. And obtaining a dark field image for each imaging, wherein the dark field image comprises a plurality of light spot areas formed by the corresponding nanorods.
And (4): the nanorods were controlled to rotate in the same direction at preset angle increments and multiple dark field images of the AuNRs in this region were recorded by dark field microscopy.
As an embodiment of this embodiment, the polarization direction of the first polarizer in the horizontal plane is the first direction. As shown in fig. 3, the horizontal angle of the nanorods refers to an angle Φ formed between the nanorods and a reference axis, wherein the reference axis is a horizontal axis constructed on the plane of the glass slide in parallel with the first direction of the first polarizing plate, i.e., when the horizontal angle is 0 °, the long axis of the nanorods is parallel with the polarization direction of the incident linearly polarized light. The horizontal angle phi of the nano rod on the plane of the glass slide is changed every 5 degrees, and the nano rod returns to the initial position after rotating 360 degrees.
The step (4) of controlling the nanorods to rotate in the same direction according to the preset angle increment can be to synchronously rotate the first polarizing film and the second polarizing film, so that the first direction of the first polarizing film is changed, and then the horizontal angle of the nanorods is changed, or the glass slide is rotated through the objective table, so that the nanorods rotate by the angle, and then the horizontal angle of the nanorods is changed.
After step S201, S202 is executed: and determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods.
The embodiment adopts polarization-induced scattering imaging, namely, the initial illumination light beam is linearly polarized, and scattered light generated by the nanorods based on the linearly polarized light is imaged. In a plurality of dark field images obtained based on polarization, under the action of linearly polarized light, the intensities of light spots formed when the same nanorod is in different directions are different. The intensity of the scattered light of the nanorods can indicate the direction of the nanorods in polarization modulation, and the intensity change of the scattered light can be obtained by imaging with a dark-field microscope, so the present embodiment can determine the orientation of each AuNRs by using polarization-excited scattering imaging.
The mapping relationship between the light intensity value and the horizontal angle of the nanorod can be represented by fitting a cosine function, specifically, a function y which represents the change of the light intensity value y with the horizontal angle phi of the nanorod is pre-established as cos (phi). After the dark field images are obtained, the light spot area in each dark field image can be obtained, the light intensity values of the light spot areas at the same position in the obtained multiple dark field images (such as 72 images) are analyzed, and the horizontal angle corresponding to the light intensity values is determined from the pre-constructed cosine function, so that the direction of the nanorod corresponding to each light spot in the dark field images is determined.
The light intensity value of the light spot area is determined by the average value of the light intensity values of all the pixel points in the light spot area or the maximum light intensity value in the light spot area. The dark field image is an image in an RGB format, each pixel point has a corresponding RGB value, the dark field image in the RGB format is converted into an image in an HSI format, wherein H represents chromaticity, S represents saturation, and I represents brightness, and the brightness value I of each pixel point in the image in the HSI format is used as a corresponding light intensity value. Can be used forOptionally, it can also be obtained by the formula
The light intensity value of the pixel is obtained by direct calculation, R, G, B represents the RGB value of the pixel.
Optionally, after step S201, that is, multiple dark-field images of the nanorods at different horizontal angles are obtained, and the light spot regions in the dark-field images are obtained by extraction as follows. Referring to fig. 4, the method includes the following steps:
s301: the dark field image is converted into a grayscale image, and the grayscale image is converted into a binary image by threshold segmentation.
After the dark field image is converted into the gray image, the processing device firstly determines the light intensity value of each pixel point, and converts the gray image into a binary image according to a preset segmentation threshold under the condition of uniform illumination, namely when the light intensity values on each pixel point in the dark field image are consistent (for example, a variance is calculated for all the light intensity values, and the light intensity values are considered to be consistent when the variance is smaller than the threshold). Under the condition of non-uniform illumination, namely when the light intensity value of each pixel point in the dark field image is inconsistent (for example, when the variance is not smaller than the threshold value, the light intensity value is considered to be inconsistent), the gray image is processed by adopting a fuzzy c-means algorithm to obtain a corrected image with uniform illumination, and the corrected image is converted into a binary image according to a preset segmentation threshold value.
In this embodiment, the preset segmentation threshold may be determined by an automatic threshold detection method, for example, by identifying a grayscale image or a corrected image by a maximum inter-class variance method and automatically obtaining the segmentation threshold.
S302: and clustering each pixel point in the binary image through a region growing algorithm to obtain a plurality of light spot regions.
After obtaining a plurality of light spot areas in each dark field image, calculating the light intensity value of each light spot area according to the RGB value of each pixel point in the light spot area, determining the direction of each nanorod according to the light intensity value, and completing the analysis of the dark field image.
Optionally, in fig. 2, after step S202, that is, obtaining the direction of each nanorod in the dark-field image, the method further includes:
s203: and generating a reconstructed image according to the direction of each nanorod.
In the present embodiment, a reconstructed image representing the position and direction of each nanorod is generated with the direction of the nanorod at the corresponding position in the dark-field image as the long axis direction, the first preset length as the long axis length of the nanorod, and the second preset length as the short axis length of the nanorod. The positions and orientations of all nanorods can be directly observed from the generated reconstructed image.
Wherein the first preset length is greater than the second preset length. In the reconstructed image, each nanorod is generated according to the same long axis length and the same short axis length, and therefore, all nanorods in the image are drawn to the same size. In the process of preparing the gold nanorods in the steps (1) and (2), the gold nanorods are uniform as much as possible, so that the expression of the generated reconstructed image can be more accurate.
Thousands of nanorods can be arranged on one dark field image or one reconstructed image, and after the direction of each nanorod is obtained, the statistical analysis of the nanorods in different directions is facilitated. Because no method can obtain the orientation of each nanorod in a nanorod sample, and the analysis of the nanorods is performed based on light spots in a dark field image, the influence of whether the direction of the nanorods has on the aspects of the speed of the photocatalytic reaction, the number of gain and loss electrons and the like in the process of generating the photocatalytic reaction cannot be considered, and therefore the system and the method in the embodiment have important significance for the experimental study of the nanorods. In practical application, when dark field analysis is performed, the method described in this embodiment can further accurately perform analysis in the same direction, and the accuracy and stability of data can be improved.
An application of the direction obtaining method provided in this embodiment is described below, with reference to fig. 5, the application includes the following steps:
s401: and acquiring target images of the nanorods at different moments, which are acquired by a dark field microscope, wherein the target images are generated after a catalyst is added into a slide bearing the nanorods.
The glass slide is provided with a deposition groove, a certain volume of AuNRs sample is deposited in the deposition groove, and a photocatalyst is added into the deposition groove and can catalyze the AuNRs to generate a photocatalytic reaction. The photocatalyst is, for example, P-mercaptoaniline (P-ATP).
S402: and determining the change information of the nano rod in the photocatalytic reaction process according to the plurality of target images at different moments.
Under the action of linearly polarized light emitted by the first polarizer and a photocatalyst, thermal electrons are transferred to acceptor molecules of an adjacent structure through Local Surface Plasmon Resonance (LSPR) excitation of the plasma metal nanorods, and photochemical conversion of the surfaces of the nanorods is promoted. In the process, the scattered light of AuNRs is changed, so that the red shift of the spectrum is caused, the RGB color of the light spot imaged in a dark field microscope is changed, and the intensity of the scattered light is obviously reduced due to the loss of electrons on the surface of the nano-rods. Therefore, by analyzing the change of the color of the light spot or the change of the scattered light intensity in the dark field image, the speed and the degree of the photocatalytic reaction can be determined, and the oxidation process in the photocatalytic reaction can be monitored.
The change information of the nanorods in the photocatalytic reaction process in the embodiment includes but is not limited to: the speed of the photocatalytic reaction, the number of electrons lost, whether the catalytic oxidation-reduction reaction occurs, and the like. Specifically, according to the change of the color of the same light spot, the specific change of the spectrum can be estimated, and further the quantity of the electron gain and loss can be quantitatively estimated.
In this embodiment, statistics is performed on the variation information of the nanorods in different directions, for example, the nanorods horizontally and vertically arranged are selected to study the variation information in the photocatalytic reaction process according to the reconstructed image.
The inventor conducts research and analysis on the light intensity change of AuNRs in all directions, and the light intensity change value of the AuNRs in the horizontal direction is similar to that of the AuNRs in the vertical direction in the same time interval, which means that the photocatalytic activity of the AuNRs is independent of the arrangement direction of the nanorods. The above findings of the inventor have guiding significance for the subsequent experimental study of the nanorods.
Compared with the prior art, the system and the method for acquiring the direction of the plasma nanorod can accurately estimate the direction of the gold nanorod (AuNRs), the scattering light intensity of the AuNRs is measured by using a dark-field microscope, and the orientation of the AuNRs is disclosed. By utilizing the anisotropic optical property of plasma scattering, the AuNRs sample can be rotated relative to linear polarization illumination under the effects of dark field microscopy and cross polarization, and only a part of AuNRs can be selectively imaged to obtain a dark field scattering image of the rotated sample under the linear polarization illumination. By utilizing a digital image processing technology, the position and direction information of AuNRs is extracted from a series of polarized dark field images, and an integral light scattering image is reconstructed, which has important significance for the analysis of a complex nanorod sample and can promote the application of the plasma nanorod in the research of rapid chemical reaction and biological process. Compared with the scanning electron microscope technology, the method is more convenient and time-saving, and observation can be carried out under any conditions. The method can further be used to monitor chemical reactions and provide highly repeatable and reliable results.
Based on the same inventive concept, an embodiment of the present application further provides a direction obtaining apparatus for plasma nanorods, please refer to fig. 6, the apparatus includes:
the image acquisition module 501 is configured to acquire a plurality of dark-field images of nanorods acquired by a dark-field microscope at different horizontal angles, where each dark-field image includes a plurality of light spot regions formed by the corresponding nanorods;
the direction determining module 502 is configured to determine a direction of each nanorod in the dark-field image according to the light intensity values of the spot areas at the same position in the multiple dark-field images and the mapping relationship between the light intensity values and the horizontal angles of the nanorods.
Optionally, the direction determining module 502 is specifically configured to: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the variation of the light intensity value with the horizontal angle of the nanorod, and the function is a cosine function.
Optionally, the apparatus further comprises: and the image reconstruction module is used for generating a reconstructed image for representing the position and the direction of each nanorod by taking the direction of the nanorod at the corresponding position in the dark field image as the long axis direction, taking the first preset length as the long axis length of the nanorod and taking the second preset length as the short axis length of the nanorod.
Optionally, the apparatus further comprises: the light spot extraction module is used for converting the dark field image into a gray level image and converting the gray level image into a binary image through threshold segmentation, wherein under the condition that the light intensity value of each pixel point in the dark field image is inconsistent, the binary image is obtained by processing the gray level image through a fuzzy c-means algorithm to obtain a corrected image with uniform illumination and then performing threshold segmentation on the corrected image; and clustering each pixel point in the binary image through a region growing algorithm to obtain a plurality of light spot regions.
Optionally, the apparatus further comprises: the reaction analysis module is used for acquiring target images of the nanorods at different moments, which are acquired by a dark-field microscope, wherein the target images are images generated in the dark-field microscope after a catalyst is added into a glass slide bearing the nanorods and the nanorods generate a photocatalytic reaction based on the catalyst; and determining the change information of the nano rod in the photocatalytic reaction process according to the plurality of target images at different moments.
The implementation principle and the generated technical effect of the direction obtaining device of the plasma nanorods provided in the embodiments of the present application are the same as those of the method embodiments described above, and for brief description, corresponding contents in the method embodiments described above may be referred to where no embodiment is mentioned in part of the embodiments of the device, and are not repeated herein.
Embodiments of the present application also provide a storage medium, which stores a computer program, and when the computer program is executed by a processor, the method for obtaining the direction of the plasma nanorods as provided in the above embodiments of the present application is performed.
The embodiment of the present application further provides a processing apparatus, which includes a processor and a memory, where the memory stores at least one instruction, at least one program, a code set, or an instruction set, and the at least one instruction, the at least one program, the code set, or the instruction set is loaded and executed by the processor, so as to implement the direction obtaining method for the plasma nanorods, provided by the above embodiment. The processing device may further comprise a communication bus, wherein the processor and the memory communicate with each other via the communication bus. The memory may include high-speed random access memory (as a cache) and may also include non-volatile memory, such as at least one magnetic disk storage device, flash memory device, or other volatile solid-state storage device. A communication bus is a circuit that connects the described elements and enables transmission between the elements. For example, the processor receives commands from other elements through the communication bus, decodes the received commands, and performs calculations or data processing according to the decoded commands.
The processing device can acquire a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles, and the direction of each nanorod in the dark field images is acquired according to the light intensity values of the light spot regions at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods. The processing device may include, but is not limited to, a desktop computer, a notebook computer, or other computing device having image processing capabilities and data analysis capabilities.
In the embodiments provided in the present application, it should be understood that the disclosed apparatus and method may be implemented in other ways. The above-described embodiments of the apparatus are merely illustrative, and for example, the division of the units is only one logical division, and there may be other divisions when actually implemented, and for example, a plurality of units or components may be combined or integrated into another system, or some features may be omitted, or not executed. In addition, the shown or discussed mutual coupling or direct coupling or communication connection may be an indirect coupling or communication connection of devices or units through some communication interfaces, and may be in an electrical, mechanical or other form.
In addition, units described as separate parts may or may not be physically separate, and parts displayed as units may or may not be physical units, may be located in one place, or may be distributed on a plurality of network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the solution of the embodiment.
Furthermore, the functional modules in the embodiments of the present application may be integrated together to form an independent part, or each module may exist separately, or two or more modules may be integrated to form an independent part.
It should be noted that the functions, if implemented in the form of software functional modules and sold or used as independent products, may be stored in a computer readable storage medium. Based on such understanding, the technical solution of the present application or portions thereof that substantially contribute to the prior art may be embodied in the form of a software product stored in a storage medium and including instructions for causing a computer device (which may be a personal computer, a server, or a network device) to execute all or part of the steps of the method according to the embodiments of the present application. And the aforementioned storage medium includes: various media capable of storing program codes, such as a usb disk, a removable hard disk, a Read-Only Memory (ROM), a Random Access Memory (RAM), a magnetic disk, or an optical disk.
In this document, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions.
The above description is only an example of the present application and is not intended to limit the scope of the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (8)
1. A directional acquisition system of plasma nanorods, comprising: the device comprises an illumination light source, a first polaroid, a second polaroid, a dark-field microscope and a processing device, wherein the illumination light source is opposite to an objective lens of the dark-field microscope, an objective table of the dark-field microscope carrying nanorods is arranged between the illumination light source and the objective lens, the first polaroid is arranged between the illumination light source and the objective table, the second polaroid is arranged between the objective table and the objective lens, and the polarization direction of the first polaroid is always perpendicular to that of the second polaroid;
the illumination light source is used for providing an initial illumination light beam;
the processing device is used for acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles, and determining the direction of each nanorod based on the dark field images, wherein each dark field image is obtained by changing the included angle between the nanorod and a horizontal axis, and the horizontal axis is parallel to the polarization direction of the first polarizer;
the processing device is also used for acquiring a plurality of dark field images of the nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot regions formed by the corresponding nanorods;
determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods;
determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot regions at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods, including: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the change of the light intensity value along with the horizontal angle of the nano rod, and the function is a cosine function;
after acquiring a plurality of dark field images of the nanorods acquired by the dark field microscope under different horizontal angles, the method further comprises the following steps:
converting the dark field image into a gray level image, and converting the gray level image into a binary image through threshold segmentation, wherein under the condition that the light intensity value of each pixel point in the dark field image is inconsistent, the binary image is obtained by processing the gray level image through a fuzzy c-means algorithm to obtain a corrected image with uniform illumination and then performing threshold segmentation on the corrected image;
and clustering each pixel point in the binary image through a region growing algorithm to obtain a plurality of light spot regions.
2. The system of claim 1, wherein the nanorods are carried on a slide, and the slide is carried on a stage of a dark field microscope; the stage rotates the slide at preset angular increments.
3. The system of claim 1, further comprising: the first polaroid and the second polaroid are fixed on the adjusting device; the adjusting device is used for controlling the first polaroid and the second polaroid to synchronously rotate according to preset angle increment.
4. A direction-finding method of plasma nanorods, applied to a processing device in the direction-finding system as claimed in any one of claims 1-3, the method comprising:
acquiring a plurality of dark field images of nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot regions formed by the corresponding nanorods;
determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods;
determining the direction of each nanorod in the dark field image according to the light intensity values of the light spot regions at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods, including: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the variation of the light intensity value with the horizontal angle of the nanorod, and the function is a cosine function.
5. The method of claim 4, wherein after determining the orientation of each nanorod in the dark-field image, the method further comprises:
and generating a reconstructed image for representing the position and the direction of each nanorod by taking the direction of the nanorod at the corresponding position in the dark-field image as the long axis direction, taking the first preset length as the long axis length of the nanorod and taking the second preset length as the short axis length of the nanorod.
6. The method of claim 4, wherein after determining the orientation of each nanorod in the dark-field image, the method further comprises:
acquiring target images of nanorods acquired by a dark-field microscope at different moments, wherein the target images are images generated in the dark-field microscope after a catalyst is added into a glass slide bearing the nanorods and the nanorods generate a photocatalytic reaction based on the catalyst;
and determining the change information of the nano rod in the photocatalytic reaction process according to the plurality of target images at different moments.
7. The method of claim 4, wherein the light intensity value of each pixel in the speckle region is formulated
Calculating to obtain the light intensity value of the light spot region, wherein the light intensity value of the light spot region is determined by the mean value of the light intensity values of all pixel points in the region or the maximum light intensity value in the region; r, G, B respectively represent the RGB values of the corresponding pixels.
8. A direction-finding device of plasma nanorods, characterized by a processing device configured in the direction-finding system of any one of claims 1-3, the device comprising:
the image acquisition module is used for acquiring a plurality of dark field images of the nanorods acquired by a dark field microscope under different horizontal angles, wherein the dark field images comprise a plurality of light spot areas formed by the corresponding nanorods;
the direction determining module is used for determining the direction of each nanorod in the dark field images according to the light intensity values of the light spot areas at the same position in the dark field images and the mapping relation between the light intensity values and the horizontal angles of the nanorods;
the direction determining module is specifically configured to: determining a horizontal angle corresponding to the light intensity value from a pre-established function, wherein the horizontal angle is used as the direction of the corresponding nanorod; wherein the function represents the variation of the light intensity value with the horizontal angle of the nanorod, and the function is a cosine function.
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