CN113267252A - Staring type confocal microscopic morphology spectrum four-dimensional detection system - Google Patents

Staring type confocal microscopic morphology spectrum four-dimensional detection system Download PDF

Info

Publication number
CN113267252A
CN113267252A CN202110532184.6A CN202110532184A CN113267252A CN 113267252 A CN113267252 A CN 113267252A CN 202110532184 A CN202110532184 A CN 202110532184A CN 113267252 A CN113267252 A CN 113267252A
Authority
CN
China
Prior art keywords
unit
light
sample
lens
fluorescence
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202110532184.6A
Other languages
Chinese (zh)
Inventor
何赛灵
罗晶
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang University ZJU
Original Assignee
Zhejiang University ZJU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang University ZJU filed Critical Zhejiang University ZJU
Priority to CN202110532184.6A priority Critical patent/CN113267252A/en
Publication of CN113267252A publication Critical patent/CN113267252A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/28—Investigating the spectrum
    • G01J3/2823—Imaging spectrometer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B11/00—Measuring arrangements characterised by the use of optical techniques
    • G01B11/24—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures
    • G01B11/25—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures by projecting a pattern, e.g. one or more lines, moiré fringes on the object
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/0208—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using focussing or collimating elements, e.g. lenses or mirrors; performing aberration correction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/021—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using plane or convex mirrors, parallel phase plates, or particular reflectors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/28—Investigating the spectrum
    • G01J3/2803—Investigating the spectrum using photoelectric array detector
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/255—Details, e.g. use of specially adapted sources, lighting or optical systems
    • 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
    • G01N21/4738—Diffuse reflection, e.g. also for testing fluids, fibrous materials
    • 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
    • G01N21/4738—Diffuse reflection, e.g. also for testing fluids, fibrous materials
    • G01N21/474—Details of optical heads therefor, e.g. using optical fibres
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G02B21/0048—Scanning details, e.g. scanning stages scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361—Optical details, e.g. image relay to the camera or image sensor

Landscapes

  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pathology (AREA)
  • Optics & Photonics (AREA)
  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

The invention discloses a staring confocal micro-topography 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, wherein the excitation unit is used for exciting a focus point of a user; 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 measurement fields of in-situ microbial detection, industrial sample microstructure detection and the like.

Description

Staring type confocal microscopic morphology spectrum four-dimensional detection system
Technical Field
The invention belongs to the technical field of optics, and particularly relates to a staring confocal micro-topography 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.
Disclosure of Invention
In order to overcome the problems in the prior art, the invention discloses a staring type confocal micro-topography 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 invention has the beneficial effects 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 invention takes the problems of the loss of three-dimensional shape recovery of hyperspectral imaging under microscopic detection and single detection mode into consideration, and acquires the three-dimensional shape data, fluorescence or reflectance map data of a sample to be detected by the confocal probe principle. 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 is further elucidated with reference to the drawing.
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 as preferred examples of the present invention, and do not limit the concept and scope of the present invention, and various changes and modifications made to the technical solution of the present invention by those skilled in the art without departing from the design concept of the present invention 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 front 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 microscopic imaging unit according to claim 1, characterized in that the image plane of the microscopic imaging unit is conjugated to a fluorescence or reflectance image plane.
4. The confocal three-dimensional topography detection system of claim 1, wherein the displacement stage in the front optical unit is replaced with a liquid zoom lens for fast zooming and increasing the scanning speed of the system.
CN202110532184.6A 2021-05-17 2021-05-17 Staring type confocal microscopic morphology spectrum four-dimensional detection system Pending CN113267252A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110532184.6A CN113267252A (en) 2021-05-17 2021-05-17 Staring type confocal microscopic morphology spectrum four-dimensional detection system

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110532184.6A CN113267252A (en) 2021-05-17 2021-05-17 Staring type confocal microscopic morphology spectrum four-dimensional detection system

Publications (1)

Publication Number Publication Date
CN113267252A true CN113267252A (en) 2021-08-17

Family

ID=77231039

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110532184.6A Pending CN113267252A (en) 2021-05-17 2021-05-17 Staring type confocal microscopic morphology spectrum four-dimensional detection system

Country Status (1)

Country Link
CN (1) CN113267252A (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113959363A (en) * 2021-09-24 2022-01-21 江苏锐精光电研究院有限公司 Line scanning type spectrum confocal device based on high-dispersion cylindrical lens and cylindrical concave grating
CN114460020A (en) * 2022-01-30 2022-05-10 清华大学深圳国际研究生院 Hyperspectral scanning system and method based on digital micro-reflector
WO2023082374A1 (en) * 2021-11-09 2023-05-19 浙江大学 Gaze-type fast hyperspectral pulse laser radar system
CN117456222A (en) * 2023-08-31 2024-01-26 台州安奇灵智能科技有限公司 Hyperspectral microscopic imaging rapid identification, classification and counting method for mixed bacteria
CN117991487A (en) * 2024-03-01 2024-05-07 瑆科仪器(广州)有限公司 Line scanning confocal optical imaging system
CN118465996A (en) * 2024-07-09 2024-08-09 北京攸维医疗科技有限公司 Polarization confocal imaging method and system

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113959363A (en) * 2021-09-24 2022-01-21 江苏锐精光电研究院有限公司 Line scanning type spectrum confocal device based on high-dispersion cylindrical lens and cylindrical concave grating
WO2023082374A1 (en) * 2021-11-09 2023-05-19 浙江大学 Gaze-type fast hyperspectral pulse laser radar system
CN114460020A (en) * 2022-01-30 2022-05-10 清华大学深圳国际研究生院 Hyperspectral scanning system and method based on digital micro-reflector
CN114460020B (en) * 2022-01-30 2023-11-17 清华大学深圳国际研究生院 Hyperspectral scanning system and method based on digital micro-reflector
CN117456222A (en) * 2023-08-31 2024-01-26 台州安奇灵智能科技有限公司 Hyperspectral microscopic imaging rapid identification, classification and counting method for mixed bacteria
CN117991487A (en) * 2024-03-01 2024-05-07 瑆科仪器(广州)有限公司 Line scanning confocal optical imaging system
CN118465996A (en) * 2024-07-09 2024-08-09 北京攸维医疗科技有限公司 Polarization confocal imaging method and system

Similar Documents

Publication Publication Date Title
CN113267252A (en) Staring type confocal microscopic morphology spectrum four-dimensional detection system
CN103439254B (en) A kind of point pupil confocal laser Raman spectra test method and device
CN102768015B (en) Fluorescence response follow-up pinhole microscopic confocal measuring device
CN104567674B (en) Bilateral fitting confocal measurement method
JP2003526814A (en) Microscope with automatic focusing device suitable for high-throughput screening
CN108267445A (en) Three-dimensional two-photon mating plate is micro- and spectrum multi-modal imaging device and method
CN102818768A (en) Multifunctional biomedical microscope
CN108120702A (en) A kind of super resolution fluorescence lifetime imaging method and device based on parallel detecting
Loerke et al. Super-resolution measurements with evanescent-wave fluorescence-excitation using variable beam incidence
CN202814861U (en) Multifunctional biomedical microscope
CN108051909A (en) A kind of extended focal depth micro imaging system of combination optical tweezer function
CN110960198A (en) Near-infrared two-region confocal microscopic imaging system based on multi-dimensional adjusting frame
EP1848983B1 (en) Fluorescence correlation microscopy with real-time alignment readout
CN104697967B (en) High-space resolution laser twin shaft confocal spectroscopic mass spectrum micro imaging method and device
JP2009540346A (en) Interference confocal microscope
CN107478332B (en) Annular light beam confocal longitudinal high-resolution imaging device
CN207440383U (en) A kind of extended focal depth micro imaging system of combination optical tweezer function
CN109187491A (en) Postposition is divided pupil differential confocal Raman, LIBS spectrum micro imaging method and device
CN106770154B (en) Space self-focusing laser differential confocal Raman spectrum detection method and device
CN214951799U (en) Staring type confocal microscopic morphology spectrum four-dimensional detection system
KR20150146074A (en) Multi-modal microscope
CN104567676B (en) Bilateral fitting differential confocal measurement method
CN216898783U (en) Confocal micro-measuring apparatu of line scanning spectrum
CN214011030U (en) Multi-mode microscopic hyperspectral imager
CN109211874A (en) Postposition is divided pupil confocal laser Raman spectra test method and device

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination