CN111043985A - Microscopic 3D (three-dimensional) morphology measuring device and method based on time-sharing multispectral image - Google Patents
Microscopic 3D (three-dimensional) morphology measuring device and method based on time-sharing multispectral image Download PDFInfo
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Abstract
The invention relates to a microscopic 3D appearance measuring device and method based on time-sharing multispectral images, wherein the device comprises: the device comprises a polychromatic light source, a longitudinal dispersion enhanced optical imaging module, a filter wheel, an image sensor and an image display and analysis module, wherein the filter wheel is provided with a plurality of narrow-band optical filters and can be used for separating the polychromatic light source entering the longitudinal dispersion enhanced optical imaging module to form monochromatic light sources with different spectral bands; the image sensor is used for collecting the gray level images of the samples under a plurality of different central wavelength spectral bands in a time-sharing manner. The device can rapidly acquire the microscopic 3D morphology of the sample under the condition of no mechanical motion. The method can realize microscopic 3D shape measurement with high transverse resolution, high longitudinal measurement precision and large longitudinal measurement range in millimeter magnitude.
Description
Technical Field
The invention belongs to the field of optical microscopic imaging, and relates to a microscopic 3D morphology measuring device and method based on time-sharing multispectral images.
Background
The existing microscopic three-dimensional topography measuring technology has a plurality of defects, such as small observation range of scanning type microscopic measurement and low environment anti-interference capability; the interferometric method requires a large number of axial scans, which limits the measurement efficiency of the method; the traditional confocal microscopic measurement single-point mechanical scanning is difficult to realize real-time and rapid three-dimensional measurement; although the parallel confocal measurement technology realizes simultaneous detection of sampling points on the same confocal section, longitudinal scanning or mechanical scanning of auxiliary equipment is still required, and the measurement efficiency and the measurement accuracy are limited to a certain extent by starting and stopping the scanning process for many times and vibration of the mechanical scanning. For example, the conventional application No. cn201510922156.x solves the problem of layer-by-layer longitudinal scanning in the conventional parallel confocal measurement technology, but still needs to acquire sample gray level images before and after the focal plane is focused, and there is mechanical start-stop motion. Therefore, there is a need for an optical measurement method to solve the above problems and achieve high-precision and high-efficiency measurement of microscopic three-dimensional features.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a microscopic 3D appearance measuring device and a microscopic 3D appearance measuring method based on time-sharing multispectral images. The device and the microscopic 3D morphology measurement method based on the time-sharing multispectral image implemented on the device can realize the acquisition of the multispectral image by time sharing under the condition of no mechanical motion and the combination of the pre-calibrated time-sharing multispectral gray difference ID(x, y) and a longitudinal height ZnAnd the relation curve is used for rapidly acquiring the microscopic 3D morphology with high transverse resolution, high longitudinal measurement precision and large longitudinal measurement range in millimeter order.
The purpose of the invention is realized as follows:
the microscopic 3D morphology measuring device based on the time-sharing multispectral image comprises a polychromatic light illumination module and a longitudinal dispersion enhanced optical imaging module.
The compound color light illumination module is sequentially provided with the following components in the light path propagation direction: the device comprises a polychromatic light source, a condenser, a filter wheel, a uniform collimating light lens group, a semi-reflecting and semi-transmitting spectroscope and an objective lens with longitudinal dispersion.
The longitudinal dispersion enhanced optical imaging module is sequentially provided with the following components in the light path propagation direction: the device comprises a sample stage with adjustable longitudinal height, an objective lens with longitudinal dispersion, a semi-reflecting and semi-transmitting spectroscope, a tube lens with longitudinal dispersion, an image sensor and an image display and analysis module.
The polychromatic light illumination module and the longitudinal dispersion enhanced optical imaging module share a semi-reflecting semi-transparent spectroscope and an objective lens with longitudinal dispersion.
The longitudinal dispersion enhanced optical imaging module comprises at least one objective lens with longitudinal dispersion, or comprises at least one tube lens with longitudinal dispersion, or other beneficial combinations.
The longitudinal dispersion enhanced optical imaging module has different focal lengths or image distances for optical signals of different wave bands, namely, under the same object distance, the object clear imaging axial positions of different wave bands are different, and the object surface height can be reversely deduced according to the clear imaging wave bands of the multispectral image sensor.
The longitudinal dispersion enhanced optical imaging module needs to eliminate transverse dispersion.
The filter wheel is provided with N (N is more than or equal to 2) narrow-band filters, and the central wavelengths of the wave bands are respectively lambda1，λ2，…，λnThe multi-color light source entering the longitudinal dispersion enhanced optical imaging module can be subjected to color separation to form single-color light sources with different spectral wave bands.
The image sensor is used for time-sharing acquisition of a sample gray level image I under N (N is more than or equal to 2) different central wavelength spectral bandsn(X, Y), X is more than or equal to 0 and less than or equal to X, Y is more than or equal to 0 and less than or equal to Y, N is more than or equal to 1 and less than or equal to N, wherein X is the total row number of the spectrum gray image, and Y is the total column number of the spectrum gray image.
The microscopic 3D morphology measuring method based on the time-sharing multispectral image, which is realized on the microscopic 3D morphology measuring device based on the time-sharing multispectral image, comprises the following steps:
The microscopic 3D morphology measurement method based on the time-sharing multispectral image can also comprise a time-sharing multispectral gray difference ID(x, y) and a longitudinal height ZnThe relation curve calibration method comprises the following operation steps:
step 5.1, adjusting the sample stage with the adjustable longitudinal height to obtain different central wavelengths lambda in a time-sharing manner1，λ2，…，λnAxial characteristic curve I of axial light intensity and defocusing amount of spectral bandλn，1≤n≤N；
Step 5.2, carrying out axial characteristic curve I on axial light intensity and defocusing amount of different central wavelength spectrum wave bandsλnCarrying out normalization processing;
step 5.3, with wave band lambda1，λ2For example, the actual operation is not limited to λ1，λ2The gray level images of different central wavelength spectrum wave bands are subjected to difference processing Iλ1-Iλ2Obtaining the wave band lambda1，λ2Differential curve ID；
Step 5.4, to differential curve IDThe linear region is subjected to linear function fitting to obtain the time-sharing multispectral gray difference IDAnd a longitudinal height ZnCalibration curve of the relationship.
The microscopic 3D morphology measurement method based on the time-sharing multispectral image can further comprise the step of correcting uneven illumination light of an optical imaging system, wherein the correction processing mode is mainly realized by dividing the obtained gray difference or differential curve by the sum of gray values of corresponding points of the two-waveband gray image.
The microscopic 3D morphology measurement method based on the time-sharing multispectral image can further comprise the step of correcting uneven sample reflectivity, wherein the correction processing mode is mainly realized by multiplying the gray value of the gray image of the spectral band with different central wavelengths obtained in the step by a relative reflection coefficient. The relative reflection coefficient is set to 1 based on the substance with the largest surface reflectivity, and the relative reflection coefficient of other substances is the substance highest reflectivity divided by the substance reflectivity. The gray value of the gray image of the substances with the same height and different reflectivity under the same wave band can be equal through the correction processing mode.
The microscopic 3D morphology measurement method based on the time-sharing multispectral image can further comprise the step of compensating the gray levels of different spectral band images in the time-sharing multispectral gray image imaging process due to the fact that the transmittance of a filter changes with the band, the illumination light intensity changes with the band, or the quantum effect of a multispectral camera changes with the band, wherein the compensation processing mode is mainly realized by multiplying the gray values of the different central wavelength spectral band gray images obtained in the step by a band adjusting coefficient. The band adjustment coefficient obtaining mode is as follows: under the illumination of a uniform multi-color light source in a specific given space, a multi-spectral camera is adopted to obtain N spectral images from a surface-flattened sample with uniform reflectivity for different wave bands, the gray scale adjustment coefficient of the maximum gray scale image is 1, and the gray scale adjustment coefficients of other N-1 wave band images are the gray scale mean value of the maximum gray scale image divided by the gray scale mean value of the wave band image.
Compared with the existing microcosmic 3D topography measurement technology, the invention adopts non-contact scanning, non-single-point scanning or layer-by-layer scanning, can realize the collection of the multispectral gray image by time sharing under the condition of no mechanical movement and combines the time-sharing multispectral gray difference I calibrated in advanceD(x, y) and a longitudinal height ZnAnd the microscopic 3D morphology with high transverse resolution, high longitudinal measurement precision and large longitudinal measurement range in millimeter order can be rapidly obtained by the relation curve.
Drawings
Fig. 1 is a schematic structural diagram of a microscopic 3D topography measuring device based on time-sharing multispectral images.
In the figure: the device comprises a 1-polychromatic light source, a 2-condenser, a 3-filter wheel, a 4-uniform light illuminating lens group, a 5-semi-reflecting and semi-transmitting spectroscope, a 6-objective lens with longitudinal dispersion, a 7-longitudinal height-adjustable sample stage, an 8-tube lens with longitudinal dispersion, a 9-image sensor and a 10-image display and analysis module.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings. It should be understood that the detailed description and specific examples, while indicating the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
Detailed description of the preferred embodiment
The present embodiment is an apparatus embodiment.
The structural schematic diagram of the microscopic 3D topography measuring device based on the time-sharing multispectral image according to the embodiment is shown in fig. 1 of the specification, and the device comprises a polychromatic light illumination module and a longitudinal dispersion enhanced optical imaging module.
The compound color light illumination module is sequentially provided with the following components in the light path propagation direction: the device comprises a polychromatic light source 1, a condenser 2, a filter wheel 3, a uniform collimated light lens group 4, a semi-reflecting and semi-transmitting spectroscope 5 and an objective lens 6 with longitudinal dispersion.
The longitudinal dispersion enhanced optical imaging module is sequentially provided with the following components in the light path propagation direction: the device comprises a sample stage 7 with adjustable longitudinal height, an objective 6 with longitudinal dispersion, a semi-reflecting semi-transmitting spectroscope 5, a tube lens 8 with longitudinal dispersion, an image sensor 9 and an image display and analysis module 10.
The polychromatic light illumination module and the longitudinal dispersion enhanced optical imaging module share a semi-reflecting semi-permeable spectroscope 5 and an objective lens 6 with longitudinal dispersion.
The longitudinal dispersion enhanced optical imaging module comprises at least one objective 6 with longitudinal dispersion, or comprises at least one tube mirror 8 with longitudinal dispersion, or other beneficial combinations.
The longitudinal dispersion enhanced optical imaging module has different focal lengths or image distances for optical signals of different wave bands, namely, under the same object distance, the object clear imaging axial positions of different wave bands are different, and the object surface height can be reversely deduced according to the clear imaging wave bands of the multispectral image sensor.
The longitudinal dispersion enhanced optical imaging module needs to eliminate transverse dispersion.
The filter wheel 3 is provided with N (N is more than or equal to 2) narrow-band filters, and the central wavelengths of the wave bands are respectively lambda1，λ2，…，λnThe multi-color light source entering the longitudinal dispersion enhanced optical imaging module can be subjected to color separation to form single-color light sources with different spectral wave bands.
The image sensor 9 is used for collecting the gray level image I of the sample under N (N is more than or equal to 2) different central wavelength spectral bands in a time-sharing wayn(X, Y), X is more than or equal to 0 and less than or equal to X, Y is more than or equal to 0 and less than or equal to Y, N is more than or equal to 1 and less than or equal to N, wherein X is the total row number of the spectrum gray image, and Y is the total column number of the spectrum gray image.
Detailed description of the invention
This embodiment is an embodiment of a method implemented on the apparatus described in the first embodiment.
The microscopic 3D morphology measuring method based on the time-sharing multispectral image comprises the following steps:
The microscopic 3D topography measuring method based on the time-sharing multispectral image further comprises a time-sharing multispectral gray difference ID(x, y) and a longitudinal height ZnThe relation curve calibration method comprises the following operation steps:
step 5.1, adjusting the sample stage 7 with the adjustable longitudinal height to obtain different central wavelengths lambda in a time-sharing manner1，λ2，…，λnAxial characteristic curve I of axial light intensity and defocusing amount of spectral bandλn，1≤n≤N；
Step 5.2, carrying out axial characteristic curve I on axial light intensity and defocusing amount of different central wavelength spectrum wave bandsλnCarrying out normalization processing;
step 5.3, with wave band lambda1，λ2For example, the actual operation is not limited to λ1，λ2Performing difference processing I on gray level images of different center wavelength spectrum wave bands lambda 1 and lambda 2λ1-Iλ2Obtaining the wave band lambda1，λ2Differential curve ID；
Step 5.4, to differential curve IDThe linear region is subjected to linear function fitting to obtain the time-sharing multispectral gray difference IDAnd a longitudinal height ZnCalibration curve of the relationship.
The microscopic 3D topography measuring method based on the time-sharing multispectral image according to this embodiment may further include performing correction processing on the unevenness of the illumination light of the optical imaging system, where the correction processing is mainly implemented by dividing the obtained gray-scale difference or differential curve by the sum of the gray-scale values of the corresponding points of the two-band gray-scale image.
The microscopic 3D topography measuring method based on the time-sharing multispectral image of the embodiment may further include correcting the unevenness of the reflectance of the specimen, wherein the correcting method is mainly implemented by multiplying the gray values of the spectral band gray images with different central wavelengths obtained in the above steps by a relative reflectance. The relative reflection coefficient is set to 1 based on the substance with the largest surface reflectivity, and the relative reflection coefficient of other substances is the substance highest reflectivity divided by the substance reflectivity. The gray value of the gray image of the substances with the same height and different reflectivity under the same wave band can be equal through the correction processing mode.
The microscopic 3D topography measurement method based on the time-sharing multispectral image in this embodiment may further include performing compensation processing on the gray levels of the different spectral band images in the time-sharing multispectral gray image imaging process due to the fact that the transmittance of a filter changes with the band, or the intensity of illumination light changes with the band, or the quantum effect of the multispectral camera changes with the band, where the compensation processing mode is mainly implemented by multiplying the gray values of the different central wavelength spectral band gray images obtained in the above steps by a band adjustment coefficient. The band adjustment coefficient obtaining mode is as follows: under the illumination of a uniform multi-color light source in a specific given space, a multi-spectral camera is adopted to obtain N spectral images from a surface-flattened sample with uniform reflectivity for different wave bands, the gray scale adjustment coefficient of the maximum gray scale image is 1, and the gray scale adjustment coefficients of other N-1 wave band images are the gray scale mean value of the maximum gray scale image divided by the gray scale mean value of the wave band image.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (8)
1. A microscopic 3D appearance measuring device based on time-sharing multispectral images is characterized in that: comprises a polychromatic light illumination module and a longitudinal dispersion enhanced optical imaging module,
the compound color light illumination module is sequentially provided with the following components in the light path propagation direction: a polychromatic light source (1), a condenser (2), a filter wheel (3), a uniform collimating light lens group (4), a semi-reflecting and semi-transmitting spectroscope (5) and an objective lens (6) with longitudinal dispersion,
the longitudinal dispersion enhanced optical imaging module is sequentially provided with the following components in the light path propagation direction: a longitudinal height adjustable sample stage (7), an objective lens (6) with longitudinal dispersion, a semi-reflecting semi-transmitting spectroscope (5), a tube lens (8) with longitudinal dispersion, an image sensor (9) and an image display and analysis module (10),
the polychromatic light illumination module and the longitudinal dispersion enhanced optical imaging module share the semi-reflecting semi-transparent spectroscope (5) and the objective lens (6) with longitudinal dispersion;
the filter wheel (3) is provided with N pieces of narrow-band filters, and the central wavelengths of the wave bands are respectively lambda1，λ2，…，λnThe multi-color light source entering the longitudinal dispersion enhanced optical imaging module can be subjected to color separation to form single-color light sources with different spectral wave bands;
the image sensor (9) is used for collecting the gray level images I of the samples under N different central wavelength spectral bands in a time-sharing mannern(X, Y), wherein N is more than or equal to 2, X is more than or equal to 0 and less than or equal to X, Y is more than or equal to 0 and less than or equal to Y, N is more than or equal to 1 and less than or equal to N, X is the total row number of the spectrum gray image, and Y is the total column number of the spectrum gray image.
2. The microscopic 3D topography measuring device based on time-shared multispectral images as claimed in claim 1, wherein: the longitudinal dispersion enhanced optical imaging module comprises at least one objective lens (6) with longitudinal dispersion or at least one tube lens (8) with longitudinal dispersion.
3. The microscopic 3D topography measuring device based on time-shared multispectral images as claimed in claim 1, wherein: the longitudinal dispersion enhanced optical imaging module needs to eliminate transverse dispersion.
4. A microscopic 3D morphology measurement method based on time-sharing multispectral images is characterized by comprising the following steps: use of the microscopic 3D topography measuring device based on time-shared multispectral images according to any of the claims 1 to 3, comprising the following operative steps:
step 1, a sample to be detected is placed on a sample table (7) with adjustable longitudinal height;
step 2, adjusting the sample stage (7) with the adjustable longitudinal height to enable the central wave band of monochromatic light to be lambdanWhen the system is used, the spectrum gray level image of the sample collected by the image sensor (9) is clear;
step 3, obtaining the central wave band of the monochromatic light as lambda in a time-sharing way through the image sensor (9)1，λ2，…，λnTime N sample spectrum gray level image In(X, Y), wherein N is more than or equal to 2, X is more than or equal to 0 and less than or equal to X, Y is more than or equal to 0 and less than or equal to Y, N is more than or equal to 1 and less than or equal to N, X is the total row number of the spectrum gray image, and Y is the total column number of the spectrum gray image;
step 4, performing difference processing on each point of the gray level image under the adjacent spectral wave bands, and performing time-sharing multispectral image gray level difference ID(x,y)＝In(x,y)-In-1(x,y)；
Step 5, time-sharing multispectral gray difference I calibrated in advanceDAnd a longitudinal height ZnAnd (4) calculating a reduction sample surface topography Z (x, y) according to the relation curve.
5. The method according to claim 4, wherein the microscopic 3D topography measurement method based on time-sharing multispectral images comprises: the method may further comprise time-sharing multispectral gray-scale difference ID(x, y) and a longitudinal height ZnThe relation curve calibration method comprises the following operation steps:
step 5.1, adjusting the sample stage (7) with the adjustable longitudinal height to obtain different central wavelengths lambda in a time-sharing manner1，λ2，…，λnAxial characteristic curve I of axial light intensity and defocusing amount of spectral bandλn，1≤n≤N；
Step 5.2, carrying out axial characteristic curve I on axial light intensity and defocusing amount of different central wavelength spectrum wave bandsλnCarrying out normalization processing;
step 5.3, different center wavelength spectrum wave bands lambda are processed1，λ2The gray level image is subjected to difference processing Iλ1-Iλ2ObtainingWave band lambda1，λ2Differential curve ID；
Step 5.4, to differential curve IDThe linear region is subjected to linear function fitting to obtain the time-sharing multispectral gray difference IDAnd a longitudinal height ZnCalibration curve of the relationship.
6. The microscopic 3D topography measuring method based on time-sharing multispectral images according to claim 4 or 5, wherein: and the correction processing mode is realized by dividing the obtained gray difference or differential curve by the sum of the gray values of the corresponding points of the gray images of the two wave bands.
7. The microscopic 3D topography measuring method based on time-sharing multispectral images according to claim 4 or 5, wherein: and correcting the uneven reflectivity of the samples in a manner of multiplying the gray values of the obtained gray images of different central wavelength spectral bands by a relative reflection coefficient.
8. The microscopic 3D topography measuring method based on time-sharing multispectral images according to claim 4 or 5, wherein: the method also comprises the step of compensating the gray levels of the images in different spectral bands in the time-sharing multispectral gray level image imaging process due to the fact that the transmittance of a filter changes with the band, the intensity of illumination light changes with the band, or the quantum effect of a multispectral camera changes with the band, wherein the compensation processing mode is realized by multiplying the gray values of the obtained gray levels of the images in different central wavelength spectral bands by a band adjusting coefficient.
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