CN216117309U - Biological microscopic vision pre-focusing device based on spectrum confocal principle - Google Patents
Biological microscopic vision pre-focusing device based on spectrum confocal principle Download PDFInfo
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Abstract
The utility model discloses a biological microscopic vision pre-focusing device based on a spectrum confocal principle. The device comprises a spectrum confocal sensing module, a micro-optical module, a slide clamping device, a focusing movement device and a scanning movement device. The spectrum confocal sensing module comprises a dispersion lens, a wide spectrum light source, an optical fiber light path, a spectrum measurement module and a control system; the microscopic optical module comprises a microscope objective, an imaging lens, a multiplying power conversion lens, a condenser lens, a light source and a digital camera device; the slide clamping device is used for fixing the standard biological slide and ensuring that the standard biological slide does not slide when moving horizontally; the focusing motion device is used for driving the slide clamping device and the microscope objective to move mutually; and the scanning motion device is used for driving the slide clamping device to move in an X-Y plane so as to complete the scanning of the standard biological slide. The utility model is used for automatic focusing of the biological digital microscope, and has the characteristics of high focusing speed, accurate layered focus prediction and the like.
Description
Technical Field
The utility model belongs to the field of automatic focusing, and particularly relates to a biological microscopic vision pre-focusing device based on a spectrum confocal principle. The utility model can be used for biological microscopic camera shooting-based equipment and instruments, and is particularly suitable for quick pre-focusing of a digital section scanner.
Background
The digital slice scanner scans and collects the traditional pathological glass slices by using a digital camera system to obtain a high-resolution digital Image for displaying the cell tissue condition of a patient, automatically carries out high-precision multi-view seamless splicing and processing on the obtained Image, and obtains a high-quality full-slice digital Image (WSI). By sharing the WSI data, remote pathological diagnosis and interpretation communication can be realized, and the sharing and utilization of high-quality medical expert resources are facilitated.
The imaging quality of digital slice scanners relies on high precision focus control, and therefore the autofocus technique of the microscopic vision system is of paramount importance. In order to improve efficiency, the conventional digital slice scanner selects some feature points on the slide, focuses the feature points to obtain the focal position, and then fits the feature focal points to a focal plane, which is also called a "focal topographic map". This process is called "pre-focusing". An autofocus microscope based on a liftable automatic stage is disclosed in the ZL200820169109.8 patent, but rapid high frequency response focusing cannot be achieved due to the high weight of the stage. Focusing was achieved by moving the image splitter lens and the microscope objective lens in patent application 03136023.8 filed by olympus optics, japan. An automatic focusing device based on a mobile object stage and a slice scanning device based on the automatic focusing device are disclosed in a patent of 'a digital slice real-time scanning automatic focusing tracking method' (ZL 201310549338.8) filed by Miaodi group of industries and Co., Ltd and a patent application 201410008180.8.
In the above techniques, feature points are focused by an image pickup device, and a "focused topographic map" is fitted. These techniques suffer from two significant drawbacks: firstly, the characteristic points are not properly selected, the phenomenon of imaging blurring of a large area is easy to occur, and the possibility of influencing diagnosis exists; secondly, the process of searching the focus needs to drive the focusing movement device to move, synchronously acquire images and calculate the position of the focus, and occupies a large amount of time. The first disadvantage is complex in cause, and focusing errors can be caused by insufficient details of the under-lens view under the characteristic points or the existence of foreign matters in non-tissue areas, which is difficult to avoid; the second disadvantage is that the pre-focus takes almost one third of the total scan time. Therefore, developing a new pre-focusing technique is necessary for further development of digital slice scanners.
Disclosure of Invention
The utility model aims to provide a faster and more accurate biological microscopic vision pre-focusing technology. The utility model is based on the spectrum confocal principle, realizes the rapid determination of the focus position by measuring the thickness of the cover glass and the tissue layer, has the characteristics of high focusing speed, high focusing precision and the like, and is particularly suitable for the focusing application of a digital pathological section scanning system.
In order to achieve the purpose, the utility model adopts the following technical scheme to solve the problem:
biological microscopic vision pre-focusing device based on spectrum confocal principle, including spectrum confocal sensing module, micro-optical module, slide clamping device, focus telecontrol equipment and scanning telecontrol equipment, wherein:
the spectrum confocal sensing module consists of a dispersion lens, a wide spectrum light source, an optical fiber light path, a spectrum measurement module and a control system; the microscopic optical module consists of a microscope objective, an imaging lens, a multiplying power conversion lens, a condenser lens, a light source and a digital camera device; the slide clamping device is used for fixing the standard biological slide and ensuring that the standard biological slide does not slide when moving horizontally; the focusing motion device is used for driving the slide clamping device and the microscope objective to move mutually; and the scanning motion device is used for driving the slide clamping device to move in an X-Y plane so as to complete the scanning of the standard biological slide.
The measuring range of the spectrum confocal sensing module is preferably between 0.1mm and 1.5mm, and most preferably 0.4 mm; the longitudinal resolution of the spectrum confocal sensing module is less than 0.2 μm, and is preferably 25 nm; the dispersion lens is used for respectively focusing light with different wavelengths to realize axial dispersion of polychromatic light; the wide-spectrum light source consists of a white light LED and a collimating lens and generates multi-wavelength composite light; the optical fiber optical path consists of a Y-shaped optical fiber, a light guide optical fiber and an optical fiber connector; the spectrum measurement module comprises a first lens, a grating light splitter, a second lens and a light detection sensor;
the Y-shaped optical fiber is provided with three ports, namely an optical inlet, an optical outlet, a beam merging port and the like to form a small-hole confocal structure, the optical inlet of the Y-shaped optical fiber is connected to the wide-spectrum light source, the optical outlet of the Y-shaped optical fiber is connected to the optical inlet of the spectrum measuring device, and the beam merging port of the Y-shaped optical fiber is connected to one end of the optical fiber connector; the other end of the optical fiber connector is connected with a light guide optical fiber, and the other end of the light guide optical fiber is connected with the dispersion lens;
white light emitted by the broad spectrum light source sequentially passes through the light inlet of the Y-shaped optical fiber, the optical fiber connector and the light guide optical fiber, enters the dispersion lens, is divided into lights with different wavelengths, and is respectively focused at different axial positions; light focused on the upper surface of the cover glass, the lower surface of the cover glass, the upper surface of the tissue and the lower surface of the tissue is reflected to enter the dispersion lens, sequentially passes through the light guide optical fiber, the optical fiber connector and the beam combining port of the Y-shaped optical fiber and then enters the light inlet of the spectrum measuring device from the light outlet of the Y-shaped optical fiber; the light entering the spectrum measuring device is collimated by the first lens and then enters the grating light splitter; the grating light splitter reflects incident light at different angles according to different wavelengths, and the incident light forms a dispersed light beam through the second lens and finally enters the optical detection sensor; the light detection sensor is a linear array detector, and can adopt a CMOS (complementary metal oxide semiconductor) or CCD (charge coupled device) image sensor;
the control system is the prior mature technology and comprises a logic control circuit, a light intensity driving and controlling circuit, a digital signal processor and the like; the logic control circuit controls the light detection sensor to acquire spectral information; the light intensity driving and controlling circuit controls the wide-spectrum light source to work and adjusts the luminous intensity of the wide-spectrum light source; and the digital signal processor converts the spectral information into the distance between the upper surface and the lower surface of the cover glass and the thickness information of the biological tissue layer to be observed.
The utility model also provides a digital section scanner based on the device and the method, and the focusing motion device comprises a Z-direction piezoelectric motion platform, a spectrum confocal sensing module, a plane reflecting mirror and a piezoelectric drive controller. The planar reflector is arranged on the movable side of the Z-direction piezoelectric motion platform and is used for reflecting dispersed light emitted by the spectrum confocal sensing module; the stroke of the Z-direction piezoelectric motion platform is smaller than the measuring range of the spectrum confocal sensing module; and the piezoelectric driving controller utilizes the displacement information of the plane reflector acquired by the spectrum confocal sensing module to carry out closed-loop focusing motion control.
Compared with the prior art, the utility model has the beneficial effects that:
first, using the principle of spectral confocal, "focal topography" can be acquired quickly by planar scanning. In the process, the visual field under the mirror is not required to be imaged, the focusing device is not required to be driven to move, and the frequency of the acquired data can reach more than 1 kHz. Therefore, the technology can greatly improve the pre-focusing efficiency.
Secondly, the focusing position covering the whole cover glass area can be collected in a short time, the reflection intensity can be reflected through the amplitude of the spectrum peak, and the impurity area can be obviously distinguished. Statistical analysis of a larger number of foci also makes it easier to eliminate erroneous foci. And the position accuracy obtained by spectral confocal is an order of magnitude higher than that of the conventional method. Therefore, the method can improve the accuracy of the focusing topographic map and ensure the definition of the image.
Drawings
FIG. 1 is a schematic diagram of the biological microscopic vision pre-focusing device based on the spectrum confocal principle according to the present invention;
FIG. 2 is a schematic diagram of an alternative of a biological microscopic vision pre-focusing device based on the spectrum confocal principle;
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings and examples.
The device comprises a spectrum confocal sensing module, a micro-optical module, a slide clamping device, a focusing movement device and a scanning movement device. The spectrum confocal sensor measures the distance to the upper surface/lower surface of the cover glass and the thickness of a biological tissue layer to be observed at one time; the spatial relationship between the object focus of the microscope objective and the measured value of the spectral confocal sensor is determined through calibration, and then the measured value of the spectral confocal sensor directly drives the focusing movement device to realize the accurate automatic focusing of biological microscopic vision. The utility model also provides a digital slice scanner based on the device and the method. The method is suitable for the construction of the focusing topographic map of the digital section scanner, can also be used for the automatic focusing of the biological digital microscope, and has the characteristics of high focusing speed, accurate layered focus prediction and the like.
The embodiment of the utility model relates to a novel device and a novel method for realizing biological microscopic vision pre-focusing based on a spectrum confocal principle, which can be used for focusing application of a digital section scanner and also can be used for automatic focusing of a biological digital microscope.
Fig. 1 shows an embodiment of a biological microscopic vision pre-focusing device based on the spectral confocal principle, which includes a spectral confocal sensing module, a microscopic optical module, a slide holding device 114, a focusing movement device 115, and a scanning movement device 116, wherein:
in this embodiment, the spectrum confocal sensing module includes a dispersive lens 101, a wide spectrum light source 102, an optical fiber light path 103, a spectrum measurement module 107, and a control system 106; the microscopic optical module comprises a microscopic objective lens 108, an imaging lens 109, a multiplying power conversion mirror 110, a condenser lens 111, a light source 112 and a digital camera device 113; the slide clamping device 114 is used for fixing the standard biological slide and ensuring that the standard biological slide does not slide when moving horizontally; the focusing motion device 115 is used for driving the slide holding device 114 and the microscope objective 108 to move mutually; the scanning motion device 116 is used to drive the slide holding device 114 to move in the X-Y plane to complete the scanning of the standard biological slide.
In this embodiment, the slide holding device 114 is mounted on the focusing motion device 115, and the focusing motion device 115 is mounted on the scanning motion device 116.
In this embodiment, the range of the spectral confocal sensing module is preferably between 0.1mm and 1.5mm, and most preferably 0.4 mm; the longitudinal resolution of the spectrum confocal sensing module is less than 0.2 μm, and is preferably 25 nm; the dispersion lens 101 is used for focusing light with different wavelengths respectively to realize axial dispersion of polychromatic light; the broad spectrum light source 102 consists of a white light LED and a collimating lens and generates multi-wavelength composite light; the optical fiber light path 103 consists of a Y-shaped optical fiber 103-1, a light guide optical fiber 103-3 and an optical fiber connector 103-2; the spectrum measuring module comprises a first lens 107-1, a grating light splitter 107-2, a second lens 107-3 and a light detection sensor 107-1.
In this embodiment, the Y-shaped optical fiber 103-1 has three ports, i.e., an optical inlet, an optical outlet, and a beam combining port, and forms a small-aperture confocal structure, wherein the optical inlet is connected to the broad-spectrum light source 102, the optical outlet is connected to the optical inlet of the spectrum measuring device 107, and the beam combining port is connected to one end of the optical fiber connector 103-2; the other end of the optical fiber connector 103-2 is connected with the light guide fiber 103-3, and the other end of the light guide fiber 103-3 is connected with the interface terminal 101-1 of the dispersion lens 101.
In this embodiment, the white light emitted by the broad spectrum light source 102 sequentially passes through the light inlet of the Y-shaped optical fiber 103-1, the optical fiber connector 103-2, and the light guide fiber 103-3, enters the dispersion lens 101, is divided into lights with different wavelengths, and is focused at different axial positions; the light focused on the upper surface of the cover glass, the lower surface of the cover glass, the upper surface of the tissue and the lower surface of the tissue is reflected to enter the dispersion lens 101, sequentially passes through the light guide fiber 103-3, the fiber connector 103-2 and the beam merging port of the Y-shaped fiber 103-1, and then enters the light inlet of the spectrum measuring device 107 through the light outlet of the Y-shaped fiber 103-1; the light entering the spectrum measuring device 107 is collimated by the first lens 107-1 and then enters the grating beam splitter 107-2; the grating light splitter 107-2 reflects incident light at different angles according to different wavelengths, and forms a dispersed light beam through the second lens 107-3, and finally the dispersed light beam is incident on the light detection sensor 107-4; the light detecting sensor 107-4 is a line array type detector, and may employ a CMOS (complementary metal oxide semiconductor) or CCD (charge coupled device) image sensor.
The control system 106 consists of a logic control circuit, a light intensity driving and controlling circuit and a digital signal processor; the logic control circuit controls the light detection sensor 107-1 to acquire spectral information; the light intensity driving and controlling circuit controls the wide-spectrum light source 102 to work and adjusts the light emitting intensity of the wide-spectrum light source; and the digital signal processor converts the spectral information into the distance between the upper surface and the lower surface of the cover glass and the thickness information of the biological tissue layer to be observed.
Fig. 2 shows another embodiment of a biological microscopic vision pre-focusing device based on the spectral confocal principle, which includes a spectral confocal sensing module, a microscopic optical module, a slide holding device 114, a focusing moving device 201 and a scanning moving device 116. In this embodiment, the focus motion device 115 is separate from the scan motion device 116, and the slide holding device 114 is secured to the scan motion device 116 by a height adjustment adapter 202.
The utility model also provides a digital section scanner based on the device and the method, and the focusing motion device comprises a Z-direction piezoelectric motion platform, a spectrum confocal sensing module, a plane reflecting mirror and a piezoelectric drive controller. The planar reflector is arranged on the movable side of the Z-direction piezoelectric motion platform and is used for reflecting dispersed light emitted by the spectrum confocal sensing module; the stroke of the Z-direction piezoelectric motion platform is smaller than the measuring range of the spectrum confocal sensing module; and the piezoelectric driving controller utilizes the displacement information of the plane reflector acquired by the spectrum confocal sensing module to carry out closed-loop focusing motion control.
The above-described embodiments are merely preferred embodiments of the present invention, which should not be construed as limiting the utility model. Various changes and modifications may be made by one of ordinary skill in the pertinent art without departing from the spirit and scope of the present invention. Therefore, the technical scheme obtained by adopting the mode of equivalent replacement or equivalent transformation is within the protection scope of the utility model.
Claims (5)
1. The biological microscopic vision pre-focusing device based on the spectrum confocal principle is characterized by comprising a spectrum confocal sensing module, a microscopic optical module, a slide clamping device, a focusing movement device and a scanning movement device;
the spectrum confocal sensing module comprises a dispersion lens, a wide spectrum light source, an optical fiber light path, a spectrum measurement module and a control system; the microscopic optical module comprises a microscope objective, an imaging lens, a multiplying power conversion lens, a condenser lens, a light source and a digital camera device; the slide clamping device is used for fixing the standard biological slide and ensuring that the standard biological slide does not slide when moving horizontally; the focusing motion device is used for driving the slide clamping device and the microscope objective to move mutually; and the scanning motion device is used for driving the slide clamping device to move in an X-Y plane so as to complete the scanning of the standard biological slide.
2. The biological microscopy vision pre-focusing device based on spectral confocal principle of claim 1, wherein the range of the spectral confocal sensing module is between 0.1mm and 1.5 mm; the longitudinal resolution is less than 0.2 μm.
3. The biological micro-vision pre-focusing device based on the spectral confocal principle of claim 1, wherein the dispersion lens is used for respectively focusing lights with different wavelengths to realize axial dispersion of the polychromatic lights; the wide-spectrum light source consists of a white light LED and a collimating lens and generates multi-wavelength composite light; the optical fiber optical path consists of a Y-shaped optical fiber, a light guide optical fiber and an optical fiber connector; the spectrum measurement module comprises a first lens, a grating light splitter, a second lens and a light detection sensor; the control system consists of a logic control circuit, a light intensity driving and controlling circuit and a digital signal processor; the logic control circuit controls the light detection sensor to acquire spectral information; the light intensity driving and controlling circuit controls the wide-spectrum light source to work and adjusts the luminous intensity of the wide-spectrum light source; and the digital signal processor converts the spectral information into the distance between the upper surface and the lower surface of the cover glass and the thickness information of the biological tissue layer to be observed.
4. The biological microscopy visual pre-focusing device based on the spectral confocal principle as claimed in claim 3, wherein the Y-shaped optical fiber has three ports of an optical input port, an optical output port and a beam combining port, and forms a small-hole confocal structure, the optical input port is connected to the broad-spectrum light source, the optical output port is connected to the optical input port of the spectral measurement module, and the beam combining port is connected to one end of an optical fiber connector; the other end of the optical fiber connector is connected with a light guide optical fiber, and the other end of the light guide optical fiber is connected with the dispersion lens.
5. The biological microscopy visual pre-focusing device based on the spectral confocal principle according to claim 4, wherein the white light emitted from the broad spectrum light source sequentially passes through the light inlet of the Y-shaped optical fiber, the optical fiber connector and the light guide optical fiber, enters the dispersion lens, is divided into lights with different wavelengths, and is focused at different axial positions respectively; light focused on the upper surface of the cover glass, the lower surface of the cover glass, the upper surface of the tissue and the lower surface of the tissue is reflected to enter the dispersion lens, sequentially passes through the light guide optical fiber, the optical fiber connector and the beam combining port of the Y-shaped optical fiber and then enters the light inlet of the spectrum measuring module from the light outlet of the Y-shaped optical fiber; the light entering the spectrum measuring module is collimated by the first lens and then enters the grating light splitter; the grating light splitter reflects incident light at different angles according to different wavelengths, and the incident light forms a dispersed light beam through the second lens and finally enters the optical detection sensor; the light detection sensor is a linear array detector and can adopt a CMOS or CCD image sensor.
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