CN102262151A - Method for quickly scanning slice by microscope - Google Patents
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- CN102262151A CN102262151A CN2010102312635A CN201010231263A CN102262151A CN 102262151 A CN102262151 A CN 102262151A CN 2010102312635 A CN2010102312635 A CN 2010102312635A CN 201010231263 A CN201010231263 A CN 201010231263A CN 102262151 A CN102262151 A CN 102262151A
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Abstract
The invention discloses a method for quickly scanning a slice by a microscope. The method comprises the following steps of: 1, setting a scanning region of the slice according to the size of a sample to be scanned; 2, setting a slice scanning direction; 3, moving a fully-automatic microscope to the center of the scanning region and starting to scan in a spiral path; 4, acquiring a group of slice images of the current visual field in the scanning path; and 5, finding out the next visual field required to be scanned by application of round detection and turning to the fourth step. By the method, quick acquisition of a slice image is realized, the scanning time is reduced and the running efficiency of a system is improved.
Description
Technical field
The present invention relates to a kind of fast scanning method of microscopic section, be particularly useful for using based on the section scanning of full microscope system.
Background technology
Use micrometron, in the process of sectioning image is obtained in scanning, may need to scan thousands of the visuals field and could obtain significative results.The image that how to obtain these visuals field more quickly is exactly the problem that needs solve.Traditional section scan method is the mode traverse scanning zone according to " bow " font, and carries out comparatively complicated focusing process in each visual field and just can obtain sectioning image.
Find in actual applications, if the zone that has the information spinner of the microscopic section of quite a few to be distributed in a circle or to concentrate relatively.
At the problems referred to above, and according to the characteristics of cutting into slices, the inventor has studied a kind of method, can obtain sectioning image more quickly, finishes scanning, and this case produces thus.
Summary of the invention
The object of the present invention is to provide a kind of fast scanning method of microscopic section, obtain sectioning image more quickly and finish scanning with realization.
In order to reach above-mentioned purpose, solution of the present invention is:
A kind of fast scanning method of microscopic section the steps include:
The first step, the size of foundation plan scanned samples is provided with the scanning area of section;
In second step, section direction of scanning (clockwise or counterclockwise) is set;
In the 3rd step, full microscope moves to the center of scanning area, beginning path scanning in a spiral manner;
In the 4th step, obtain one group of sectioning image of working as forward view in the scanning pattern;
The 5th step, use circular the detection and find the next visual field that needs scanning, forwarded for the 4th step to;
Described circular detection method is to judge by edge detection algorithm whether the next visual field in the scanning pattern needs scanning, promptly add up known blank visual field number around the visual field to be scanned, if reaching three width of cloth or surpass three width of cloth, the blank visual field on every side just judges that the visual field to be scanned is the blank visual field, but for the corner visual field, it is the previous visual field in the visual field to be scanned in the scanning pattern, back a visual field and this visual field are not point-blank, basis for estimation is that the blank visual field on every side reaches two width of cloth or just judges that above two width of cloth this visual field to be scanned is the blank visual field, and the blank visual field does not need to scan again.
Described spiral way is: outwards scan from the center of scanning area according to clockwise or counter clockwise direction, the scanning course is a straight line.
In obtaining scanning pattern, when one group of sectioning image of forward view, use buffer memory focal plane technology, described buffer memory focal plane technology is before moving to a visual field, precompute this visual field scanning Z axle focal plane reference position, then from this reference position, towards a direction step-scan, in calculator memory, write down the sectioning image in per step simultaneously, after collecting required sectioning image, stop mobile Z axle focal plane, last again with the image of focal plane the most clearly of buffer memory in the internal memory, and the image of the certain number of plies in front and back is stored.
Use stepping focal plane searching method in obtaining scanning pattern when one group of sectioning image of forward view, described stepping focal plane searching method is meant the focal plane searching method is divided into three types: coarse adjustment, middle mediation accurate adjustment; Coarse adjustment is to search clear focal plane in fixing absolute coordinates scope; The middle accent begins to search clear focal plane according to bigger interval steps from current location; Accurate adjustment is to begin to search clear focal plane according to less interval steps from current location; If detect sectioning image is near clear focal plane, just directly uses accurate adjustment to come automatic focus; The automatic focus if sectioning image is transferred in the just use far away of clear focal plane; If the sectioning image that continuous a plurality of visuals field can't be found to be needed in scanning pattern, occurs, just use coarse adjustment to search for focal plane.
After adopting such scheme, integrated use of the present invention spiral way scanning, circular detect, buffer memory focal plane technology and stepping focal plane searching method, outwards travel through whole scanning area in a screw fashion from the lining, use circular the detection in the scanning process, judge by edge detection algorithm whether the next visual field in the scanning pattern needs scanning, simultaneously, when scanning each visual field that need scan, utilize stepping focal plane searching method, use different focus methods at different situations, fast, stably to obtain section focal plane image; Utilize buffer memory focal plane technology, the picture rich in detail that buffer memory is scanned reduces moving stage repeatedly.That realizes sectioning image thus obtains and finishes scanning fast, and the present invention has reduced sweep time, has improved the operational efficiency of system.
Description of drawings
Fig. 1 is to use the section scanning synoptic diagram of spiral way, left figure expression scanning counterclockwise, right figure expression scanning clockwise;
Fig. 2 is to use the section scanning synoptic diagram of " bow " font mode;
Fig. 3 is the circular synoptic diagram that detects;
Fig. 4 is three kinds of automatic focus transformational relation figure.
Embodiment
Concrete implementation step of the present invention is as follows:
The first step, the size of foundation plan scanned samples is provided with the scanning area of section
In second step, the section direction of scanning is set for counterclockwise.
In the 3rd step, full microscope moves to the center of scanning area, beginning path scanning in a spiral manner;
In the 4th step, obtain one group of sectioning image of working as forward view in the scanning pattern;
The 5th step, use circular the detection and find the next visual field that needs scanning, forwarded for the 4th step to;
Described circular detection method is to judge by edge detection algorithm whether the next visual field in the scanning pattern needs scanning, promptly add up known blank visual field number around the visual field to be scanned, if reaching three width of cloth or surpass three width of cloth, the blank visual field on every side just judges that the visual field to be scanned is the blank visual field, but for the corner visual field, it is the previous visual field in the visual field to be scanned in the scanning pattern, back a visual field and this visual field are not point-blank, basis for estimation is that the blank visual field on every side reaches two width of cloth or just judges that above two width of cloth this visual field to be scanned is the blank visual field, and the blank visual field does not need to scan again.
The mode of scanning is spiral way traverse scanning zone.So-called spiral way is exactly outwards to scan from the center of scanning area according to clockwise or counter clockwise direction, and the scanning course is a straight line, sees Fig. 1.
This mode has following advantage with respect to traditional " bow " font scanning (see figure 2) mode:
One, the position that is positioned at the scanning area center nearly all is not the blank visual field, locatees clear focal plane and finds sectioning image than being easier to like this.If with the scan mode of " bow " font, might need through just finding the visual field that sectioning image is arranged behind the blank visual field of dozens of.
Two,, cut into slices as long as the scanning hundreds of is individual even tens effective field of view just can be collected abundant sectioning image for some according to the difference of specifically using.If use spiral scan mode,, just might end more early scan owing to just can traverse the blank visual field that only contains the small number of slices edge of image visual field and do not contain sectioning image at last.
Three, for the section of circular distribution, if according to the mode of " bow " font, four corners of scanning area also can be scanned.In fact these zones are blank, do not need scanning.If according to spiral way scanning, these zones just might accomplish not to be scanned, thereby reduce sweep time.
The circular method that detects is to add up known blank visual field number around the visual field to be scanned.If surpassing three width of cloth visuals field is blank (comprising three width of cloth visuals field), just judge that this visual field to be scanned is the blank visual field, but for the corner visual field (in the scanning pattern the previous visual field in the visual field to be scanned, a back visual field and this visual field not point-blank be called the corner visual field) basis for estimation is that to surpass two width of cloth visuals field (comprising two width of cloth visuals field) be that the blank visual field just judges that this visual field to be scanned is the blank visual field, the blank visual field does not need scanning.
The section that different batches is made, the position of its sample may have some difference, so the center of scanning area not necessarily just in time is sample center of distribution position.Detect the edge that to find as soon as possible that sample distributes by circle, thereby reduce scanning, reduce sweep time zone outside the edge.
Fig. 3 has described and has had the circular counterclockwise spiral scan process that detects, and this width of cloth figure is positioned at the lower left corner of scanning area, and each grid is represented the width of cloth visual field in the scanning pattern among the figure, and the line segment of band arrow is represented scanning pattern.
Make a concrete analysis of several examples below.Describe for convenient, be the level and the vertical direction numbering in the visual field, capitalization is the position of visual field horizontal direction, and numeral is the position of visual field vertical direction.
The visual field to be scanned is the E5 visual field, eight width of cloth visuals field on every side are respectively D4, D5, D6, E4, E6, F4, F5 and F6, wherein D4, D5, D6, E6 do not have the scanned visual field, according to before this sweep record as can be known, F4, F5 are the visuals field that image is arranged, other two width of cloth E4 and F6 are the blank visuals field, so this width of cloth visual field needs scanning.
The visual field to be scanned is the E6 visual field, eight width of cloth visuals field on every side are respectively D5, D6, D7, E5, E7, F5, F6 and F7, wherein D5, D6, D7, E7 do not have the scanned visual field, according to the record that scans before this as can be known, F5 is the visual field that image is arranged, other three width of cloth E5, F6 and F7 are the blank visuals field, so this width of cloth visual field do not need scanning, simultaneously this width of cloth visual field are labeled as the blank visual field.
The visual field to be scanned is the E7 visual field, eight width of cloth visuals field on every side are respectively D6, D7, D8, E6, E8, F6, F7 and F8, wherein D6, D7, D8, E8, F8 do not have the scanned visual field, according to before this sweep record as can be known, other three width of cloth E6, F6 and F7 are the blank visuals field, therefore this width of cloth visual field does not need scanning, simultaneously this width of cloth visual field is labeled as the blank visual field.
The visual field to be scanned is the E8 visual field, eight width of cloth visuals field on every side are respectively D7, D8, D9, E7, E9, F7, F8 and F9, wherein D7, D8, D9, E9, F8, F9 do not have the scanned visual field, according to before this sweep record as can be known, other two width of cloth E7 and F7 are the blank visuals field, simultaneously this width of cloth visual field is the corner visual field, so this width of cloth visual field do not need scanning, simultaneously this width of cloth visual field is labeled as the blank visual field.
The rest may be inferred, and F8 does not need scanning, G8 needs scanning, and comprehensive above the analysis is after microscope has scanned the E5 visual field, do not need scanning by E6, E7, E8 and F8 are analyzed in the visual field on the scanning pattern subsequently, microscope can directly move to the scanning of G8 position during scanning.
Buffer memory focal plane technology is before moving to a visual field, precomputes this visual field scanning Z axle focal plane reference position.From this reference position,, in calculator memory, write down the sectioning image in per step simultaneously then towards a direction step-scan.After collecting required sectioning image, stop mobile Z axle focal plane.Last again with the image of focal plane the most clearly of buffer memory in the internal memory, and the image of the certain number of plies in front and back is stored.
Because microscopic section has certain thickness, therefore the multilayer focal plane image that might need to obtain the same visual field just can obtain required information.Traditional method is heuristic search, and this method at first moves up and down objective table, finds the position of clear focal plane, and then removes to collect sectioning image according to both direction up and down.Classic method generally needs to move 5～6 positions of focal plane just can finish search.And the method for buffer memory focal plane does not need only to need to guarantee to pass focal plane and get final product towards a scanning direction towards the moving stage repeatedly of 2 directions up and down.Buffer memory focal plane method is general only to need 3～4 positions of focal plane just can finish.So just can reduce quite a few sweep time.
Because section is not desirable surface level at microscopically, so when microscope moves to a visual field, not always on focal plane the most clearly.In addition, situations such as the initial visual field, the adjacent visual field, fringe field of view, the blank visual field all need focus method targetedly.The search of stepping focal plane is meant the focal plane searching method is divided into three types: coarse adjustment, middle mediation accurate adjustment, different type application is to different scenes.Coarse adjustment is to search clear focal plane in fixing absolute coordinates scope; The middle accent begins to search clear focal plane according to bigger interval steps from current location; Accurate adjustment is to begin to search clear focal plane according to less interval steps from current location.If detect sectioning image is near clear focal plane, just directly uses accurate adjustment to come automatic focus.The automatic focus if sectioning image is transferred in the just use far away of clear focal plane.If the sectioning image that continuous a plurality of visuals field can't be found to be needed in scanning pattern, occurs, just use coarse adjustment to search for focal plane.Simultaneously, these three kinds of focal plane search-type can transform (see figure 4) mutually.
Claims (4)
1. the fast scanning method of a microscopic section is characterized in that step is:
The first step, the size of foundation plan scanned samples is provided with the scanning area of section;
In second step, the section direction of scanning is set;
In the 3rd step, full microscope moves to the center of scanning area, beginning path scanning in a spiral manner;
In the 4th step, obtain one group of sectioning image of working as forward view in the scanning pattern;
The 5th step, use circular the detection and find the next visual field that needs scanning, forwarded for the 4th step to;
Described circular detection method is to judge by edge detection algorithm whether the next visual field in the scanning pattern needs scanning, promptly add up known blank visual field number around the visual field to be scanned, if reaching three width of cloth or surpass three width of cloth, the blank visual field on every side just judges that the visual field to be scanned is the blank visual field, but for the corner visual field, it is the previous visual field in the visual field to be scanned in the scanning pattern, back a visual field and this visual field are not point-blank, basis for estimation is that the blank visual field on every side reaches two width of cloth or just judges that above two width of cloth this visual field to be scanned is the blank visual field, and the blank visual field does not need to scan again.
2. a kind of fast scanning method of microscopic section according to claim 1 is characterized in that: described spiral way is according to clockwise or counterclockwise outwards scan from the center of scanning area, and the scanning course is a straight line.
3. a kind of fast scanning method of microscopic section according to claim 1, it is characterized in that: in obtaining scanning pattern, when one group of sectioning image of forward view, use buffer memory focal plane technology, described buffer memory focal plane technology is before moving to a visual field, precompute scanning Z axle focal plane reference position, then from this reference position, towards a direction step-scan, in calculator memory, write down the sectioning image in per step simultaneously, after collecting required sectioning image, stop mobile Z axle focal plane, last again with the image of focal plane the most clearly of buffer memory in the internal memory, and the image of the certain number of plies in front and back is stored.
4. a kind of fast scanning method of microscopic section according to claim 1, it is characterized in that: use stepping focal plane searching method in obtaining scanning pattern when one group of sectioning image of forward view, described stepping focal plane searching method is meant the focal plane searching method is divided into three types: coarse adjustment, middle mediation accurate adjustment; Coarse adjustment is to search clear focal plane in fixing absolute coordinates scope; The middle accent begins to search clear focal plane according to bigger interval steps from current location; Accurate adjustment is to begin to search clear focal plane according to less interval steps from current location; If detect sectioning image is near clear focal plane, just directly uses accurate adjustment to come automatic focus; The automatic focus if sectioning image is transferred in the just use far away of clear focal plane; If the sectioning image that continuous a plurality of visuals field can't be found to be needed in scanning pattern, occurs, just use coarse adjustment to search for focal plane.
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| CN102566023A (en) * | 2012-01-11 | 2012-07-11 | 麦克奥迪实业集团有限公司 | Digital slide real time scanning automatic focusing system and method thereof |
| CN103323940A (en) * | 2013-05-29 | 2013-09-25 | 哈尔滨工业大学 | Electric translation device based on two-photon microscope and two-dimensional spiral scanning method thereof |
| CN103323939A (en) * | 2012-03-20 | 2013-09-25 | 麦克奥迪实业集团有限公司 | Digital slide real-time scanning automatic focus system and method |
| CN104458579A (en) * | 2014-12-12 | 2015-03-25 | 宁波江丰生物信息技术有限公司 | Digital slide scanner and scanning focus number reducing method thereof |
| CN106202956A (en) * | 2016-07-20 | 2016-12-07 | 宁波江丰生物信息技术有限公司 | A kind of flow and method of the pathological information system being applicable to Urology Department |
| DE102016110988A1 (en) * | 2016-06-15 | 2017-12-21 | Sensovation Ag | Method for digitally recording a sample through a microscope |
| CN110672608A (en) * | 2019-10-15 | 2020-01-10 | 南京泰立瑞信息科技有限公司 | Dynamic planning method and system for full-slice scanning path |
| CN112067466A (en) * | 2020-07-17 | 2020-12-11 | 南京航空航天大学 | In-situ analysis device and method for in-plane shear matrix cracks of ceramic matrix composite |
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| CN110672608B (en) * | 2019-10-15 | 2022-04-12 | 南京泰立瑞信息科技有限公司 | Dynamic planning method and system for full-slice scanning path |
| CN112067466A (en) * | 2020-07-17 | 2020-12-11 | 南京航空航天大学 | In-situ analysis device and method for in-plane shear matrix cracks of ceramic matrix composite |
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