CN105004723A - Pathological section scanning 3D imaging and fusion device and method - Google Patents
Pathological section scanning 3D imaging and fusion device and method Download PDFInfo
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Abstract
The invention provides a pathological section scanning 3D imaging and fusion device and method. The device is characterized in that the device comprises a digital camera (1), an image-forming mirror (2), an object lens (3), a loading platform (4), a scanning platform (5), a collecting lens (6), a light source (7) and a computer (8), and the computer (8) is electrically connected to the digital camera (1), the object lens (3), the scanning platform (5) and the light source (7), is used for controlling the digital camera (1) and processing image data, is used for controlling a vertical motion controller of the object lens (3) so that the object lens (3) is driven, is used for controlling a horizontal motion controller of the scanning platform (5) so that the scanning platform (5) is driven, and is used for controlling the light source (7) so that brightness is adjusted. The invention also provides a pathological section scanning 3D imaging and fusion method adopting the above device.
Description
Technical field
The present invention relates to the digitalized sliced sheet field in digital pathology, particularly relate to a kind of pathological section scanning 3 D imaging and fusing device and method.
Background technology
Pathological section is the one of Pathologic specimen.During making, part there are the tissue of pathology or internal organs through the process of various chemicals and burying storage, make it fixing sclerosis, microtome is thinly sliced, stick on slide, dye, with shades of colour, supplies to check under the microscope, to observe pathological change, make pathological diagnosis, for clinical diagnosis and treatment are offered help.
Along with infotech is in the development of medical domain and application, digital pathology become pathology now and the direction of Future Development.In digital pathology, be a step that is the most basic, most critical by slice digital.Digitalized sliced sheet is that case section is carried out full visual field high resolution scanning by digital slices scanning system, by the seamless spliced process of computing machine, and real-playback of with high fidelity cutting into slices.Digitalized sliced sheet can make case Digitalization, networking, realize the permanent storage of visualized data and be not subject to the synchronous browsing of space-time restriction, to diagnose at Clinico Pathologic, pathology Scientific Research and Teaching etc. create more far-reaching influence in field, and all will play huge effect to whole medical domain.
At present, pathological section scanister is used to make slice digital become prior art.Pathological section scanister is digitized for pathological section a kind of instrument, it instead of conventional microscope, there is informationalized advantage, but, existing pathological section scanister does not have 3D effect, and cause cannot realizing can lowering some focal lengths to observe different cell tissue changes as microscope on same position, thus the demand scanning all image details clearly in a focal plane cannot be met.
Summary of the invention
In order to solve the above-mentioned problems in the prior art, the invention provides a kind of pathological section scanning 3 D imaging and fusing device and method.The present invention is intended to effectively to simulate similar microscopical operating effect by this novel pathological section scanister, thus more convenient, succinctly, intuitively watch 3D effect.And the clear tissue image in different focal planes all can also be fused on same image by fusion function by this device, thus final realization can be complete and clearly view the image of the upper all cells of section and tissue.
First aspect, the invention provides a kind of pathological section scanning 3 D imaging and fusing device, it is characterized in that, comprising: digital camera 1, for gathering sectioning image; Imaging lens 2, it is connected to below digital camera, for the imaging of light path; Object lens 3, it is connected to below imaging lens, comprises a focusing knob, a Z axis motor, a vertical movement controller, and it can move up and down along Z-direction, for enlarged image, and for digital camera focusing; Loading platform 4, it is positioned at below object lens, for loading and removing section; Scanning platform 5, it is installed on the bottom of loading platform, comprises an X-axis motor and a y-axis motor, and a tangential movement controller, and it can move along X, Y direction, for moving horizontally section; Condenser 6, it is positioned at below scanning platform, for light is flocked together; Light source 7, it is positioned at below condenser, for regulating brightness; Computing machine 8, is electrically connected in described digital camera 1, object lens 3, scanning platform 5 and light source 7; For controlling digital camera 1 and image data processing; For controlling the vertical movement controller of object lens 3, with drived control object lens 3; For the tangential movement controller of gated sweep platform 5, to drive scanning platform 5; For controlling light source 7, to regulate brightness.
Preferably, the described object lens 3 in described pathological section scanning 3 D imaging and fusing device, also comprise low range object lens and high magnification object lens.On this basis further preferably, described object lens 3 are all less than or equal to 1 μm along Z-direction repetitive positioning accuracy.Again further preferably, described scanning platform 5 is all less than or equal to 1 μm along X-axis, Y direction repetitive positioning accuracy.
Still more preferably, in described pathological section scanning 3 D imaging and fusing device, described digital camera 1 is high-speed area array digital camera, is connected with computing machine 8 by USB3.0 connecting line.On this basis further preferably, the frame frequency >=100 frame/second of described high-speed area array digital camera.
Still more preferably, in described pathological section scanning 3 D imaging and fusing device, described imaging lens 2 is three order imaging lens, and it comprises two observation eyepieces for eye-observation and a scanning eyepiece for scanning.
Still more preferably, in described pathological section scanning 3 D imaging and fusing device, the connection of described digital camera 1 and imaging lens 2 is for removably connecting.On this basis further preferably, described digital camera 1 and the connection of imaging lens 2 are that draw-in groove connects or is threaded.On this basis further preferably, described digital camera 1, imaging lens 2, object lens 3, loading platform 4, scanning platform 5, condenser 6 and light source 7, be installed on same vertical axis from top to bottom successively.
Second aspect, present invention also offers a kind of pathological section scanning 3 D imaging and fusion method, and the method use above-mentioned pathological section scanning 3 D imaging and fusing device, this device comprises: digital camera 1, for gathering sectioning image; Imaging lens 2, it is connected to below digital camera, for the imaging of light path; Object lens 3, it is connected to below imaging lens, comprises a focusing knob, a Z axis motor, a vertical movement controller, and it can move up and down along Z-direction, for enlarged image, and for digital camera focusing; Loading platform 4, it is positioned at below object lens, for loading and removing section; Scanning platform 5, it is installed on the bottom of loading platform, comprises an X-axis motor and a y-axis motor, and a tangential movement controller, and it can move along X, Y direction, for moving horizontally section; Condenser 6, it is positioned at below scanning platform, for light is flocked together; Light source 7, it is positioned at below condenser, for regulating brightness; Computing machine 8, is electrically connected in described digital camera 1, object lens 3, scanning platform 5 and light source 7; For controlling digital camera 1 and image data processing; For controlling the vertical movement controller of object lens 3, with drived control object lens 3; For the tangential movement controller of gated sweep platform 5, to drive scanning platform 5; For controlling light source 7, to regulate brightness.
Described method comprises step:
1. the microslide being loaded with section is positioned on loading platform 4, and fixing;
2. use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly;
3. survey focal length by Z axis, computing machine 8 adopts a series of images planar fit method to calculate a virtual plane, i.e. 0 layer, focal plane, and determines scan start point;
4. object lens 3 are controlled by computing machine 8 and scanning platform 5 moves, thus mobile 0 layer, described focal plane, obtain track A ', digital camera 1 captures the image in motion process in real time, and this Image Real-time Transmission to computing machine 8, computing machine 8 processes this image transmitted in real time, until the track A ' end of scan;
5. 0 layer, focal plane is moved to described scan start point again along Y-axis, and the Z axis track of 0 layer, focal plane is moved down certain distance α, and the focal length value of corresponding change Z axis, then according to step 4. run-down again; And:
6. step is repeated 4. extremely 5., the Z axis track of 0 layer, focal plane is constantly moved up or moves down certain distance α, until all layer ends of scan, thus obtain all view data of 0 layer of-n layer, the 3D rendering on A track can be depicted by all view data of this 0 layer of-n layer.
7. repeat step 3. to 6., depict B track, C track, D track successively ... thus the 3D rendering obtained on all tracks.
8. utilize all tomographic images on above-mentioned all tracks, use computing machine 8 calculate 3D data and by blending algorithm, take out every layer of content the most clearly, finally synthesize a fused layer.
Preferably, in described pathological section scanning 3 D imaging and fusion method, described step 2. in focusing be auto-focusing or manual focus.
Further preferably, in described pathological section scanning 3 D imaging and fusion method, the object lens 3 in described device also comprise low range object lens and high magnification object lens.On this basis, 2. preferred described step is: first, adopts low range object lens scan slice, transfers data to computing machine 8 by digital camera 1, and computing machine 8 display obtains preliminary panorama sketch of cutting into slices, and selectes region to be measured on this panorama sketch; Then, be converted to high magnification object lens, in the image in this region to be measured, use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly.
Still more preferably, in described pathological section scanning 3 D imaging and fusion method, described distance alpha≤1 μm.
Still more preferably, in described pathological section scanning 3 D imaging and fusion method, described planar fit method is two-dimensional interpolation method or least square method.
Use apparatus and method of the present invention, carry out pathological section scanning, can facilitate, succinctly, intuitively watch 3D effect, after the clear tissue image in different focal planes is all fused on same image by described device, experiment testing staff can also be complete and clearly view the image of the upper all cells of section and tissue, meets the demand scanning all picture rich in detail details in a focal plane simultaneously.
Accompanying drawing explanation
Fig. 1 is pathological section scanning 3 D imaging of the present invention and fusing device;
1-digital camera; 2-imaging lens; 3-object lens; 4-loading platform; 5-scanning platform; 6-condenser; 7-light source; 8-computing machine;
Fig. 2 is the X, Y, Z axis trajectory diagram of object lens and the movement of scanning platform compound;
Fig. 3 is X, the Y-axis trajectory diagram of scanning platform movement;
Fig. 4 is the Y of A ' track, the movement locus figure of Z axis.
Embodiment
Below in conjunction with embodiment, the present invention is further elaborated, but the present invention is not limited to following embodiment.
Embodiment 1
A kind of pathological section scanning 3 D imaging and fusing device
As shown in Figure 1, described device comprises: digital camera 1, for gathering sectioning image; Imaging lens 2, it is connected to below digital camera, for the imaging of light path; Object lens 3, it is connected to below imaging lens, comprises a focusing knob, a Z axis motor, a vertical movement controller, and it can move up and down along Z-direction, for enlarged image, and for digital camera focusing; Loading platform 4, it is positioned at below object lens, for loading and removing section; Scanning platform 5, it is installed on the bottom of loading platform, comprises an X-axis motor and a y-axis motor, and a tangential movement controller, and it can move along X, Y direction, for moving horizontally section; Condenser 6, it is positioned at below scanning platform, for light is flocked together; Light source 7, it is positioned at below condenser, for regulating brightness; Computing machine 8, is electrically connected in described digital camera 1, object lens 3, scanning platform 5 and light source 7; For controlling digital camera 1 and image data processing; For controlling the vertical movement controller of object lens 3, with drived control object lens 3; For the tangential movement controller of gated sweep platform 5, to drive scanning platform 5; For controlling light source 7, to regulate brightness.
Wherein, described object lens 3, also comprise low range object lens and high magnification object lens; Described object lens 3 are equal to 1 μm along Z-direction repetitive positioning accuracy; Described scanning platform 5 is equal to 1 μm along X-axis, Y direction repetitive positioning accuracy.
Wherein, described digital camera 1 is high-speed area array digital camera, is connected with computing machine 8 by USB3.0 connecting line.On this basis further preferably, the frame frequency=100 frame/second of described high-speed area array digital camera.
Wherein, described imaging lens 2 is three order imaging lens, and it comprises two observation eyepieces for eye-observation and a scanning eyepiece for scanning.
Wherein, described digital camera 1 and the connection of imaging lens 2 are for being threaded.
Embodiment 2
A kind of pathological section scanning 3 D imaging and fusing device
As shown in Figure 1, described device comprises: digital camera 1, for gathering sectioning image; Imaging lens 2, it is connected to below digital camera, for the imaging of light path; Object lens 3, it is connected to below imaging lens, comprises a focusing knob, a Z axis motor, a vertical movement controller, and it can move up and down along Z-direction, for enlarged image, and for digital camera focusing; Loading platform 4, it is positioned at below object lens, for loading and removing section; Scanning platform 5, it is installed on the bottom of loading platform, comprises an X-axis motor and a y-axis motor, and a tangential movement controller, and it can move along X, Y direction, for moving horizontally section; Condenser 6, it is positioned at below scanning platform, for light is flocked together; Light source 7, it is positioned at below condenser, for regulating brightness; Computing machine 8, is electrically connected in described digital camera 1, object lens 3, scanning platform 5 and light source 7; For controlling digital camera 1 and image data processing; For controlling the vertical movement controller of object lens 3, with drived control object lens 3; For the tangential movement controller of gated sweep platform 5, to drive scanning platform 5; For controlling light source 7, to regulate brightness.
Wherein, described object lens 3, also comprise low range object lens and high magnification object lens; Described object lens 3 are equal to 0.5 μm along Z-direction repetitive positioning accuracy; Described scanning platform 5 is equal to 0.5 μm along X-axis, Y direction repetitive positioning accuracy.
Wherein, described digital camera 1 is high-speed area array digital camera, is connected with computing machine 8 by USB3.0 connecting line.On this basis further preferably, the frame frequency=200 frame/second of described high-speed area array digital camera.
Wherein, described imaging lens 2 is three order imaging lens, and it comprises two observation eyepieces for eye-observation and a scanning eyepiece for scanning.
Wherein, described digital camera 1 is that draw-in groove connects with the connection of imaging lens 2.And described digital camera 1, imaging lens 2, object lens 3, loading platform 4, scanning platform 5, condenser 6 and light source 7, be installed on same vertical axis from top to bottom successively.
Embodiment 3
A kind of pathological section scanning 3 D imaging and fusion method
Described method employs device of the present invention (as shown in Figure 1), and comprises step:
1. the microslide being loaded with section is positioned on loading platform 4, and fixing;
2. use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly;
3. survey focal length by Z axis, computing machine 8 adopts two-dimensional interpolation method to calculate a virtual plane, i.e. 0 layer, focal plane, and determines scan start point; As shown in Figure 3,9 points in figure represent actual focusing, and A wherein represents the actual slice track while scan of X, Y-axis, i.e. A track;
4. object lens 3 are controlled by computing machine 8 and scanning platform 5 moves, thus mobile 0 layer, described focal plane, obtain track A ' (as shown in Figure 2, the wherein track of A ' indication and the curve movement of Z axis, be called for short A ' track, it is corresponding with the A track in Fig. 3), digital camera 1 captures the image in motion process in real time, and this Image Real-time Transmission to computing machine 8, computing machine 8 processes this image transmitted in real time, until the track A ' end of scan;
5. 0 layer, focal plane is moved to described scan start point again along Y-axis, and the Z axis track of 0 layer, focal plane is moved down 1 μm, and the focal length value of corresponding change Z axis, then according to step 4. run-down again; And:
6. step is repeated 4. extremely 5., the Z axis track of 0 layer, focal plane is constantly moved up or moves down 1 μm, until all layer ends of scan (Y of A ' track as shown in Figure 4, the movement locus figure of Z axis, every bar curve respectively illustrates the track of 4 layers, 0 layer, focal plane to focal plane), thus obtain all view data of 0 layer-4 layers, the 3D rendering on A track can be depicted by these all view data of 0 layer-4 layers.
7. repeat step 3. to 6., depict B track, C track, D track, E track, F track, G track successively, thus obtain the 3D rendering on all tracks.
8. utilize all tomographic images on above-mentioned all tracks, use computing machine 8 calculate 3D data and by blending algorithm, take out every layer of content the most clearly, finally synthesize a fused layer.
Embodiment 4
A kind of pathological section scanning 3 D imaging and fusion method
Described method employs device of the present invention (as shown in Figure 1), and the object lens 3 in described device also comprise low range object lens and high magnification object lens, and comprise step:
1. the microslide being loaded with section is positioned on loading platform 4, and fixing;
2. first, adopt low range object lens scan slice, transfer data to computing machine 8 by digital camera 1, computing machine 8 display obtains preliminary panorama sketch of cutting into slices, and selectes region to be measured on this panorama sketch; Then, be converted to high magnification object lens, in the image in this region to be measured, use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly.
3. survey focal length by Z axis, computing machine 8 adopts least square method to calculate a virtual plane, i.e. 0 layer, focal plane, and determines scan start point; As shown in Figure 3,9 points in figure represent actual focusing, and A wherein represents the actual slice track while scan of X, Y-axis, i.e. A track;
4. object lens 3 are controlled by computing machine 8 and scanning platform 5 moves, thus mobile 0 layer, described focal plane, obtain track A ' (as shown in Figure 2, the wherein track of A ' indication and the curve movement of Z axis, be called for short A ' track, it is corresponding with the A track in Fig. 3), digital camera 1 captures the image in motion process in real time, and this Image Real-time Transmission to computing machine 8, computing machine 8 processes this image transmitted in real time, until the track A ' end of scan;
5. 0 layer, focal plane is moved to described scan start point again along Y-axis, and the Z axis track of 0 layer, focal plane is moved down 0.5 μm, and the focal length value of corresponding change Z axis, then according to step 4. run-down again; And:
6. step is repeated 4. extremely 5., the Z axis track of 0 layer, focal plane is constantly moved up or moves down 0.5 μm, until all layer ends of scan, thus obtain all view data of 0 layer-4 layers, the 3D rendering on A track can be depicted by these all view data of 0 layer-4 layers.
7. repeat step 3. to 6., depict B track, C track, D track, E track, F track, G track successively, thus obtain the 3D rendering on all tracks.
8. utilize all tomographic images on above-mentioned all tracks, use computing machine 8 calculate 3D data and by blending algorithm, take out every layer of content the most clearly, finally synthesize a fused layer.
Be described in detail specific embodiments of the invention above, but it is just as example, the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (16)
1. section scanning 3 D imaging and a fusing device, is characterized in that, comprising:
Digital camera (1), for gathering sectioning image;
Imaging lens (2), it is connected to below digital camera, for the imaging of light path;
Object lens (3), it is connected to below imaging lens, comprises a focusing knob, a Z axis motor, a vertical movement controller, and it can move up and down along Z-direction, for enlarged image, and for digital camera focusing;
Loading platform (4), it is positioned at below object lens, for loading and removing section;
Scanning platform (5), it is installed on the bottom of loading platform, comprises an X-axis motor and a y-axis motor, and a tangential movement controller, and it can move along X, Y direction, for moving horizontally section;
Condenser (6), it is positioned at below scanning platform, for light is flocked together;
Light source (7), it is positioned at below condenser, for regulating brightness;
Computing machine (8), is electrically connected in described digital camera (1), object lens (3), scanning platform (5) and light source (7); For controlling digital camera (1) and image data processing; For controlling the vertical movement controller of object lens (3), with drived control object lens (3); For the tangential movement controller of gated sweep platform (5), to drive scanning platform (5); For controlling light source (7), to regulate brightness.
2. section scanning 3 D imaging according to claim 1 and fusing device, is characterized in that, described object lens (3) also comprise low range object lens and high magnification object lens.
3. section scanning 3 D imaging according to claim 2 and fusing device, is characterized in that, described object lens (3) are all less than or equal to 1 μm along Z-direction repetitive positioning accuracy.
4. section scanning 3 D imaging according to claim 3 and fusing device, is characterized in that, described scanning platform (5) is all less than or equal to 1 μm along X-axis, Y direction repetitive positioning accuracy.
5. section scanning 3 D imaging according to claim 4 and fusing device, is characterized in that, described digital camera (1) is high-speed area array digital camera, is connected with computing machine (8) by USB3.0 connecting line.
6. section scanning 3 D imaging according to claim 5 and fusing device, is characterized in that, the frame frequency >=100 frame/second of described high-speed area array digital camera.
7. section scanning 3 D imaging according to claim 4 and fusing device, is characterized in that, described imaging lens (2) is three order imaging lens, and it comprises two observation eyepieces for eye-observation and a scanning eyepiece for scanning.
8. the section scanning 3 D imaging according to any one of claim 1-7 and fusing device, is characterized in that, the connection of described digital camera (1) and imaging lens (2) is for removably connecting.
9. section scanning 3 D imaging according to claim 8 and fusing device, is characterized in that, described digital camera (1) and the connection of imaging lens (2) are that draw-in groove connects or is threaded.
10. section scanning 3 D imaging according to claim 8 and fusing device, it is characterized in that, described digital camera (1), imaging lens (2), object lens (3), loading platform (4), scanning platform (5), condenser (6) and light source (7), be installed on same vertical axis from top to bottom successively.
11. 1 kinds use the section scanning 3 D imaging of device as claimed in claim 1 and the method for fusion, it is characterized in that, comprise step:
1. the microslide being loaded with section is positioned on loading platform (4), and fixing;
2. use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly;
3. survey focal length by Z axis, computing machine (8) adopts a series of images planar fit method to calculate a virtual plane, i.e. 0 layer, focal plane, and determines scan start point;
4. moved by computing machine (8) control object lens (3) and scanning platform (5), thus mobile 0 layer, described focal plane, obtain track A ', digital camera (1) captures the image in motion process in real time, and this Image Real-time Transmission to computing machine (8), computing machine (8) processes this image transmitted in real time, until the track A ' end of scan;
5. 0 layer, focal plane is moved to described scan start point again along Y-axis, and the Z axis track of 0 layer, focal plane is moved down certain distance α, and the focal length value of corresponding change Z axis, then according to step 4. run-down again; And:
6. step is repeated 4. extremely 5., the Z axis track of 0 layer, focal plane is constantly moved up or moves down certain distance α, until all layer ends of scan, thus obtain all view data of 0 layer of-n layer, the 3D rendering on A track can be depicted by all view data of this 0 layer of-n layer.
7. repeat step 3. to 6., depict B track, C track, D track successively ... thus the 3D rendering obtained on all tracks.
8. utilize all tomographic images on above-mentioned all tracks, use computing machine (8) calculate 3D data and by blending algorithm, take out every layer of content the most clearly, finally synthesize a fused layer.
The method of 12. section scanning 3 D imagings according to claim 11 and fusion, is characterized in that, described step 2. in focusing be auto-focusing or manual focus.
The method of 13. section scanning 3 D imagings according to claim 12 and fusion, is characterized in that, the object lens (3) in described device also comprise low range object lens and high magnification object lens.
The method of 14. section scanning 3 D imagings according to claim 13 and fusion, it is characterized in that, 2. described step is: first, adopt low range object lens scan slice, computing machine (8) is transferred data to by digital camera (1), computing machine (8) display obtains preliminary panorama sketch of cutting into slices, and selectes region to be measured on this panorama sketch; Then, be converted to high magnification object lens, in the image in this region to be measured, use described device several focuses to image to focus, calculate the Z axis pinpointed focus of each focus, search out image position the most clearly.
The method of 15. section scanning 3 D imagings according to any one of claim 11-14 and fusion, is characterized in that, described distance alpha≤1 μm.
The method of 16. section scanning 3 D imagings according to claim 15 and fusion, is characterized in that, described planar fit method is two-dimensional interpolation method or least square method.
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