CN214951799U - Staring type confocal microscopic morphology spectrum four-dimensional detection system - Google Patents
Staring type confocal microscopic morphology spectrum four-dimensional detection system Download PDFInfo
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Abstract
The utility model discloses a staring confocal micro-morphology spectrum four-dimensional detection system, which comprises an excitation unit, a front optical unit, a micro-imaging unit, a middle scanning unit and a rear receiving unit; exciting light of the sample to be detected is led in from the excitation unit and enters the middle scanning unit; the galvanometer system of the middle scanning unit performs two-dimensional scanning, and the emergent exciting light is reflected by the beam splitter to enter the microscope objective and focus a sample to be measured; imaging the reflected light to form a conjugate microscopic image; one path of signal fluorescence or reflected light is finally emitted to an external photoelectric detector after returning, and the other path of signal fluorescence or reflected light is emitted to the hyperspectral light splitting module to form a fluorescence map or a reflection map. The system fuses the three-dimensional microscopic morphology and the fluorescence or reflection map data into four-dimensional spectral morphology microscopic data, has extremely high spectral resolution and depth resolution, and has great application value in the fields of in-situ microbial detection, industrial sample microstructure detection and the like.
Description
Technical Field
The utility model belongs to the technical field of optics, concretely relates to staring type confocal micro morphology spectrum four-dimensional detection system.
Background
At present, hyperspectral imaging is based on a multichannel spectral technology, optical imaging and spectral measurement are integrated, and image information and corresponding spectral information of a target can be acquired simultaneously. The hyperspectral imaging can analyze, measure and process the structure and the components of a substance, has the advantages of high analysis precision, wide measurement range and the like, and is widely applied to the fields of petroleum, materials, agriculture, geological exploration, biochemistry, medical sanitation, environmental protection, safety detection and the like. However, the traditional hyperspectral imaging technology can only acquire the spectral information of the object to be measured, cannot acquire the three-dimensional morphology information of the object to be measured, and cannot restore the real four-dimensional information of the object with morphology and chromaticity characteristics. The traditional three-dimensional reconstruction technology cannot acquire spectral information of an object. The microscope system can magnify and observe detailed image information of a sample to be detected, is an important tool for detecting the microscopic properties of the sample, and is a difficult point in the industry at present how to realize the integrated acquisition of the microscopic three-dimensional appearance and the spectral information of the sample to be detected. In addition, when the hyperspectral imaging technology is used for microscopic detection, most of commercial hyperspectral imagers can only work in a single detection mode and are difficult to be used for multi-mode detection of samples with different characteristics.
SUMMERY OF THE UTILITY MODEL
In order to overcome the problems in the prior art, the utility model discloses a staring type confocal micro-morphology spectrum four-dimensional detection system.
A staring confocal micro-topography spectrum four-dimensional detection system comprises an excitation unit, a preposed optical unit, a micro-imaging unit, a middle scanning unit and a rear receiving unit; the exciting unit comprises a reflector and an optical fiber collimator, exciting light of a sample to be detected is led in from the exciting unit and is incident to the middle scanning unit through the adjustment of the reflector; the middle scanning unit sequentially comprises a telecentric lens, a scanning lens, a vibrating mirror system, a first beam splitter and a filter plate, the scanning lens and the telecentric lens optimize light beams and expand the light beams, and the vibrating mirror system performs two-dimensional scanning; the preposed optical unit comprises an objective table, a microscope objective, a displacement table and a beam splitter, exciting light emitted from the middle scanning unit is reflected by the beam splitter to be incident into the microscope objective and focused on a sample to be detected on the objective table, the displacement table is used for adjusting the distance of the sample to be detected to realize focusing, and a fluorescence signal or a reflected light signal excited by the sample to be detected returns along an original light path; the microscopic imaging unit comprises a fourth focusing lens and a first camera, and light reflected by a sample to be detected is imaged at the position of the first camera through the fourth focusing lens to form a conjugate microscopic image; the rear end receiving unit comprises a second beam splitter, a first focusing lens, a pinhole, a second focusing lens and a hyperspectral light splitting module, signal fluorescence or reflected light enters the second beam splitter in the rear end receiving unit through the middle scanning unit along an original light path, is focused at the position of the pinhole through the first focusing lens and is emitted to an external photoelectric detector, and the other path of signal fluorescence or reflected light enters the hyperspectral light splitting module through the second focusing lens to form a fluorescence map or a reflection map.
The hyperspectral light splitting module sequentially comprises a slit, a collimating lens, a prism-grating-prism, a third focusing lens and a second camera, light is emitted from the middle scanning unit to the position of the slit and collimated into parallel light by the collimating lens, and after passing through the prism-grating-prism, the light with different wavelengths is focused on different positions of a light-sensitive surface of the second camera through the third focusing lens, so that a spectral image is formed.
The image surface of the microscopic imaging unit is conjugated with the fluorescence spectrum or reflection spectrum image surface.
The displacement table in the front optical unit is replaced by a liquid zoom lens for realizing rapid zooming and improving the scanning speed of the system.
The utility model has the advantages that:
the confocal probe principle is adopted to detect the three-dimensional shape information and the fluorescence or reflection spectrum information of the sample to be detected. Fusing the three-dimensional shape data and the fluorescence or reflection map data of the sample to be detected into a four-dimensional data cube. The system has the characteristics of high depth resolution, high spectral resolution, excellent data fusion precision and the like. The new spectrum appearance data volume greatly improves the accuracy of microscopic detection and analysis of the substance.
The utility model discloses consider the disappearance that three-dimensional appearance that the micro-detection was right down the high spectral imaging resumes and the single problem of detection mode, gather the three-dimensional appearance data of the sample that awaits measuring, fluorescence or reflection map data through the confocal probe principle of copolymerization. The three-dimensional appearance data and the fluorescence or reflection map data of the sample to be detected are fused into the four-dimensional spectral appearance data set, so that the means and the method for detecting the object by the system are increased, and the availability and the richness of the system are greatly improved. The method has important significance for analyzing the microscopic properties of the sample to be detected, and has great application value in the measurement fields of in-situ microbial detection, industrial sample microstructure detection and the like.
Drawings
FIG. 1 is a schematic diagram of a four-dimensional detection system for a staring confocal micro-topography spectrum;
in the figure, an object stage 1, a microscope objective 2, a displacement stage 3, a beam splitter 4, a telecentric lens 5, a scanning lens 6, a galvanometer system 7, a first beam splitter 8, a filter 9, a second beam splitter 10, a first focusing lens 11, a pinhole 12, a second focusing lens 13, a hyperspectral beam splitting module 14, a slit 15, a collimating lens 16, a prism-grating-prism 17, a third focusing lens 18, a reflector 19, an optical fiber collimator 20, a fourth focusing lens 21, a first camera 22 and a second camera 23.
Detailed Description
The invention will be further elucidated with reference to the drawings.
A staring confocal micro-topography spectrum four-dimensional detection system comprises an excitation unit, a front optical unit, a micro-imaging unit, a middle scanning unit and a rear receiving unit. The system can simultaneously acquire three-dimensional shape data, fluorescence or reflection map data of a sample to be detected based on a confocal probe principle. And fusing the three-dimensional shape data and the fluorescence or reflection map data of the sample to be detected into a four-dimensional spectral shape data set. The exciting light is led into the middle scanning unit from the exciting unit, and the galvanometer system realizes scanning by changing the incident angle of the exciting light to the scanning lens. The displacement platform makes the sample be placed on the focal plane of the microscope objective by adjusting the distance between the microscope objective and the sample to be measured. A spot on the sample to be measured is excited to fluoresce or the spot produces a reflected light signal. The exciting light can be focused at different depth positions of the sample to be measured, only the fluorescence signal on the focal plane of the microscope objective lens in the front optical unit can be collected by the rear receiving unit, and the fluorescence signal of the non-focal plane can be blocked by the confocal pinhole in the rear receiving unit. The sample to be detected can realize the fluorescence map imaging of different depths through tomography.
The confocal micro-topography detection is to continuously change the distance between a displacement table of a front optical unit and a sample to be detected, and position the axial distance between the sample to be detected and a microscope objective lens through the maximum value of a focused light intensity signal in an acquired rear-end receiving unit, so as to accurately obtain the accurate three-dimensional position data of the surface point of the sample to be detected. Then, fluorescence or reflection spectrum data are collected through a hyperspectral light splitting module of the rear-end receiving unit; and finally, scanning the two-dimensional points of the sample to be detected through a galvanometer system and obtaining the three-dimensional appearance of the sample to be detected and the hyperspectral fluorescence or reflectance spectrum of each space point of the sample to be detected.
As shown in fig. 1, the excitation unit includes a mirror 19 and a fiber collimator 20, and excitation light of a sample to be measured is introduced from the excitation unit and enters the central scanning unit through adjustment of the mirror 19. The system can scan the three-dimensional appearance of the sample to be measured through a confocal probe. The optical properties of the sample to be tested can be characterized by collecting fluorescence data or reflectance profile data.
The middle scanning unit sequentially comprises a telecentric lens 5, a scanning lens 6, a vibrating mirror system 7, a first beam splitter 8 and a filter plate 9, the scanning lens 6 and the telecentric lens 5 optimize light beams and expand the beams, and the vibrating mirror system 7 performs two-dimensional scanning. The galvanometer system 7 realizes scanning by changing the incident angle of the scanning lens 6, and the scanning lens 6 and the telecentric lens 5 optimize the scanning image surface and improve the diameter of light spots, so that scanning light beams with different angles are always converged at the entrance pupil of the micro objective lens through a reflector (beam splitter) of the front optical unit. The laser light is emitted from the central scanning unit to the front optical unit.
The front-mounted optical unit comprises an objective table 1, a microscope objective 2, a displacement table 3 and a beam splitter 4, exciting light emitted from the middle scanning unit is reflected by the beam splitter 4 to enter the microscope objective 2 and is focused on a sample to be detected on the objective table 1, the displacement table 3 is used for adjusting the distance of the sample to be detected to realize focusing, and a fluorescence signal or a reflected light signal excited by the sample to be detected returns along an original light path. The displacement table 3 in the front optical unit can be replaced by a liquid zoom lens to realize rapid zooming and improve the scanning speed of the system.
The microscopic imaging unit comprises a fourth focusing lens 21 and a first camera 22, and light reflected by a sample to be measured is imaged at the position of the first camera 22 through the fourth focusing lens 21 to form a conjugate microscopic image. The image surface of the microscopic imaging unit is conjugated with the image surface of the fluorescence spectrum or the reflection spectrum, so that a color microscopic image can be observed in the microscopic imaging unit, and the color microscopic image can be used for quickly focusing and positioning the position of a sample to be measured in a view field. The system fuses the three-dimensional microscopic morphology and confocal fluorescence or reflection spectrum data into a four-dimensional spectral morphology microscopic data set.
The rear end receiving unit comprises a second beam splitter 10, a first focusing lens 11, a pinhole 12, a second focusing lens 13 and a hyperspectral light splitting module 14, signal fluorescence or reflected light enters the second beam splitter 10 in the rear end receiving unit through a middle scanning unit along an original light path, is focused at the position of the pinhole 12 through the first focusing lens 11 and is emitted to an external photoelectric detector, and the other path of signal fluorescence or reflected light enters the hyperspectral light splitting module through the second focusing lens 13 to form a fluorescence map or a reflection map. The external photoelectric detector can adopt a single photoelectric detector or a double-channel photoelectric detector, if the single photoelectric detector is replaced by the double-channel photoelectric detector, the differential confocal detection is implemented, and therefore the three-dimensional reconstruction precision of the system can be further improved to a certain extent.
The hyperspectral light splitting module sequentially comprises a slit 15, a collimating lens 16, a prism-grating-prism 17, a third focusing lens 18 and a second camera 23, light is emitted from the middle scanning unit to the slit 15 and collimated into parallel light by the collimating lens 16, and after passing through the prism-grating-prism 17, light with different wavelengths is focused on different positions of a light-sensitive surface of the second camera 23 through the third focusing lens 18, so that a spectral image is formed. The system scans the position of a point to be detected on the surface of a sample to be detected through points to realize fluorescence or reflection map scanning imaging in a staring state.
The embodiments in the above description can be further combined or replaced, and the embodiments are only described in the preferred embodiments of the present invention, and are not limited to the concept and scope of the present invention, and various changes and modifications made by the technical solutions of the present invention by those of ordinary skill in the art without departing from the design concept of the present invention all belong to the protection scope of the present invention. The scope of the invention is given by the appended claims and any equivalents thereof.
Claims (4)
1. A staring type confocal micro-topography spectrum four-dimensional detection system is characterized in that: the device comprises an excitation unit, a preposed optical unit, a microscopic imaging unit, a middle scanning unit and a rear receiving unit;
the exciting unit comprises a reflector and an optical fiber collimator, exciting light of a sample to be detected is led in from the exciting unit and is incident to the middle scanning unit through the adjustment of the reflector;
the middle scanning unit sequentially comprises a telecentric lens, a scanning lens, a vibrating mirror system, a first beam splitter and a filter plate, the scanning lens and the telecentric lens optimize light beams and expand the light beams, and the vibrating mirror system performs two-dimensional scanning;
the preposed optical unit comprises an objective table, a microscope objective, a displacement table and a beam splitter, exciting light emitted from the middle scanning unit is reflected by the beam splitter to be incident into the microscope objective and focused on a sample to be detected on the objective table, the displacement table is used for adjusting the distance of the sample to be detected to realize focusing, and a fluorescence signal or a reflected light signal excited by the sample to be detected returns along an original light path;
the microscopic imaging unit comprises a fourth focusing lens and a first camera, and light reflected by a sample to be detected is imaged at the position of the first camera through the fourth focusing lens to form a conjugate microscopic image;
the rear end receiving unit comprises a second beam splitter, a first focusing lens, a pinhole, a second focusing lens and a hyperspectral light splitting module, signal fluorescence or reflected light enters the second beam splitter in the rear end receiving unit through the middle scanning unit along an original light path, is focused at the position of the pinhole through the first focusing lens and is emitted to an external photoelectric detector, and the other path of signal fluorescence or reflected light enters the hyperspectral light splitting module through the second focusing lens to form a fluorescence map or a reflection map.
2. The system of claim 1, wherein: the hyperspectral light splitting module sequentially comprises a slit, a collimating lens, a prism-grating-prism, a third focusing lens and a second camera, light is emitted from the middle scanning unit to the position of the slit and collimated into parallel light by the collimating lens, and after passing through the prism-grating-prism, the light with different wavelengths is focused on different positions of a light-sensitive surface of the second camera through the third focusing lens, so that a spectral image is formed.
3. The system of claim 1, wherein the image plane of the microscopic imaging unit is conjugate to the fluorescence or reflectance image plane.
4. The system of claim 1, wherein the displacement stage of the front optical unit is replaced by a liquid zoom lens for fast zooming and increasing the scanning speed of the system.
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