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CN102998787B - Fiber cross section section focusing digital picture acquisition methods under a kind of microscope - Google Patents

Fiber cross section section focusing digital picture acquisition methods under a kind of microscope Download PDF

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Publication number
CN102998787B
CN102998787B CN201210257990.8A CN201210257990A CN102998787B CN 102998787 B CN102998787 B CN 102998787B CN 201210257990 A CN201210257990 A CN 201210257990A CN 102998787 B CN102998787 B CN 102998787B
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China
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section
focusing
fiber cross
moving stage
computer control
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CN201210257990.8A
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CN102998787A (en
Inventor
徐鑫华
董绍伟
邰文峰
薛卫
王香香
曹锡忠
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JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
NANJING ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU OF PEOPLE'S REPUBLIC OF CHINA
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JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
NANJING ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU OF PEOPLE'S REPUBLIC OF CHINA
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Abstract

Fiber cross section section focusing digital picture acquisition methods under a kind of microscope, comprise the following steps: (1) obtains preliminary panorama sketch, for the three-dimensional moving stage related in system, record objective table surface level XY direction reference position, mobile Z-direction is to priori focusing position, start objective table after XY direction motion track and moving step length are set, the image mosaic of fixing a point to take in moving process become intact fibre to cut into slices after terminating and scheme greatly, and moving stage is to reference position; (2) for preliminary panorama sketch, calculate XY direction projection, projection minimum value position is fiber cross section centre of slice position; (3) center is converted into objective table XY direction displacement, moving stage, alignment lens centre of slice position, the two benches search strategy based on priori focusing position completes auto-focusing.The invention enables and manually saved, reduce the erroneous judgement of successive image process.

Description

Fiber cross section section focusing digital picture acquisition methods under a kind of microscope
Technical field
The present invention relates to digital image processing field, especially a kind of fiber cross section image acquiring method, specifically a kind of under the microscope of auto-focusing fiber cross section section panorama digital image acquisition method.
Background technology
At present, to be difficult to chemically after the fiber blends such as fiber crops, cotton, wool, chemical fibre or mechanical means detects its fiber parameters, now commonplace a kind of detection method is micro projection, adopts common biological microscope (enlargement factor is at 200 times ~ 250 times) or microscopic projector to observe fiber.The method specified according to China's textile industry standard gets out microslide and is placed on common biological microscope or microscopic projector objective table, observed each fiber come into view by eyepiece, differentiate its type according to the Morphologic Characteristics of fiber or by instrument computing nodes area.Require during detection that fiber samples reaches some, testing staff must move in the horizontal direction microslide, for the view crossing the visual field, and all fibres completing identification, counting or measure by eyepiece cross-hair center.After crossing each horizontal throw of the visual field, slowly be moved across the visual field in the horizontal direction again by after microslide vertically a mobile segment distance, identify and counting fiber, repeat this running program, until all microslide is finished watching, standard-required number of fiber reaches a setting value., there is the place that some can improve in process like this, first, the movement in surface level XY direction goes departure comparatively large by people, may re-treatment some fibre; The second, for xsect, need people manually to go localized fiber position and microscopical focusing; 3rd, limit by microscope CCD camera, collect an image just very little panel region of computing machine, fibre section, border cannot be seen entirely.
In order to overcome these problems, needing a kind of method obtaining clear fiber cross section section digital picture, on XYZ three-dimensional fiberscope objective table, obtaining by means of methods such as Gray Projection, two step auto-focusing the digital picture covering complete effective break area.
Summary of the invention
The object of this invention is to provide the clear fibre section digital image methods of a kind of acquisition, method adopts Gray Projection method to determine, and section fiber position and gradient focusing operator complete microscopical auto-focusing, alleviate testing staff's burden, for the testing of subsequent fiber parametric image provides digital picture.
Technical scheme of the present invention is:
Fiber cross section section focusing digital picture acquisition methods under a kind of microscope, described method adopt equipment comprise with three-dimensional moving stage microscope, be arranged on camera on microscope and computer control, described computer control is connected with the corresponding control signal end of camera, three-dimensional moving stage (adopting JTH-XX series small microscope automatic carrier JTH200XY-100), and described image acquiring method specifically comprises the following steps:
(1) three-dimensional moving stage is adopted to obtain preliminary panorama sketch: first, by computer control record three-dimensional moving stage surface level XY direction reference position, fiber cross section section is placed on three-dimensional moving stage, mobile Z-direction is to priori focusing position, adopt computer control that the motion track of three-dimensional moving stage in XY direction and moving step length are set, start objective table; Three-dimensional moving stage carries out work according to the aforementioned track that arranges, computer control starts camera and carries out fixed point shooting for the image under microscope at the moving step length pause point place of three-dimensional moving stage, after terminating, the image of fixing a point in moving process to take is sent to computer control by camera, computer control is spliced into the complete large figure of fiber cross section section, and three-dimensional moving stage moves to reference position;
(2) for preliminary panorama sketch and the large figure of complete fiber cross section section, computer control calculates fiber cross section and to cut into slices the projection of large figure in X, Y-direction, obtains the center of fiber cross section section;
(3) center is converted into objective table XY direction displacement, computer control drives three-dimensional moving stage that microscopical camera lens is moved to centre of slice position, the two benches search strategy based on priori focusing position is adopted to complete the auto-focusing of Z-direction, and repeat step (1), the fiber cross section section panorama sketch obtaining final focusing is again scanned in XY plane.
In step of the present invention (1), described priori focusing position refers to the focusing position that the previous use of three-dimensional moving stage of computer control record produces afterwards.
In step of the present invention (1), the moving step length of fixed point shooting is the lap of adjacent two width images, accounts for the 25-35% of picture traverse.
In step of the present invention (2), calculate the Gray Projection of X, Y-direction, when two direction projections are minimum value, position grouping is fiber cross section centre of slice position together.
In step of the present invention (2), calculate fibre section center by XY direction projection, projection formula is as follows:
T x ( i ) = 1 N Σ j = 1 N I ( i , j )
T y ( j ) = 1 M Σ i = 1 M I ( i , j )
Wherein, M is image X-direction number of pixels, and N is image Y-direction pixel number, I (i, j) the pixel gray scale that to be coordinate be (i, j).
Centre of slice coordinate is T x(i), T yi corresponding during (j) minimum value min, j min.
S x(i min)=min{T x(i)}
S y(j min)=min{T y(j)}
In step of the present invention (3), the auto-focusing adopting the two benches search strategy based on priori focusing position to complete Z-direction specifically comprises, first step search adopts larger objective table Z-direction step-length, adopts less step length searching when focusing operator size generation transition trend.
In step of the present invention (3), find fiber cross section centre of slice position, moving stage, microscopical object lens center is made to aim at centre of slice position, objective table completes sectioning image auto-focusing, adopt gradient operator C as image focusing evaluate operator in Z-direction one segment limit, carry out focus search, be best focusing position when C gets maximal value, operator as shown in the formula:
C ( z ) = Σ i = 1 M - 1 Σ j = 1 N - 1 ( ( I ( i + 1 , j ) - I ( i , j ) ) 2 + ( I ( i , j + 1 ) - I ( i , j ) ) 2 ) )
The span of z is the objective table Z-direction distance of swimming.
Beneficial effect of the present invention:
The present invention adopt Gray Projection method determine section fiber position and gradient focusing operator complete microscopical auto-focusing, alleviate testing staff's burden, for the testing of subsequent fiber parametric image provides digital picture.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of the inventive method.
Fig. 2 is the schematic diagram of objective table horizontal X Y-direction motion track when generating panorama sectioning image in the present invention.
Embodiment
Below in conjunction with drawings and Examples, the present invention is further illustrated.
As shown in Figure 1, 2, a kind of Fig. 1 illustrates process flow diagram of the present invention, and concrete gatherer process is as follows:
1st step, the CCD camera of general microscope configuration is all comparatively simple, cannot realize mobile fast in take pictures, therefore objective table is when moving by Fig. 2, X-direction move be not a step to the end, but segmentation is advanced, and allows camera fix the time.The distance s of every section xguarantee that the adjacent two width figure that CCD camera photographs have overlap, size is advisable with 1/4 picture traverse.When y moves in direction too, but y direction is a displacement s yput in place, also will guarantee that the adjacent two width figure of Y-direction have overlap, be advisable with 1/4 picture altitude.
Record objective table reference position, after XY direction track (Fig. 2) and moving step length are set, start objective table, the image mosaic of fixing a point to take in moving process become intact fibre to cut into slices after terminating and scheme greatly, and moving stage is to reference position.Also unrestricted to this as how splicing the present invention.
2nd step, for figure large after splicing, calculates XY direction projection, finds out fibre section center (i min, j min).
3rd step, by (i min, j min) be converted into objective table XY direction displacement (i min* s x/ M, j min* s y/ N), moving stage, alignment lens centre of slice position, completes Z-direction auto-focusing by process below:
A, judging whether that first time uses, in this way, make objective table vertical at Z() direction covers all distances of swimming, and objective table moves from from object lens farthest to the most nearby, calculates best focusing parameter, and to preserve this best focusing position be initial position z init; Otherwise transfer objective table is to initial position, and find the initial position time when first time uses longer, this also depends on the performance of objective table certainly.
B, arrange initial ranging moving step length step, move as positive dirction toward object lens direction, direction parameter is direction, and initial value is 1, maximum focusing evaluation of estimate C max=0, image counter i=0, define a storage of linked list and often walk the evaluation of estimate gathering image.
C, objective table Z-direction move direction*step, gather the i-th width image, Calculation Estimation value C i, preserve evaluation of estimate, displacement to chained list.
D, the 2nd width figure start, as C i-C i-1>=0, objective table moves with step by former direction, gets back to C step.
E otherwise, if C i-1>=C i-2, reduce moving step length to step/3, setting fine search scope is the distance that current location starts that preserve to chained list in the other direction at the i-th-2.Calculate the evaluation of estimate of each step, the point with maximal value is best focusing, upgrades initial position z simultaneously initfor new focusing position.Jump to G step.
F otherwise, direction negate, i.e. direction=-1*direction, carry out opposite direction search, get back to C step.
G, end.
4th step, gets back to objective table XY direction reference position, with the clear spliced map of the 1st step similar Resurvey intact fibre object.
The part that the present invention does not relate to prior art that maybe can adopt all same as the prior art is realized.

Claims (3)

1. fiber cross section section focusing digital picture acquisition methods under a microscope, it is characterized in that described method adopt equipment comprise with three-dimensional moving stage microscope, be arranged on camera on microscope and computer control, described computer control is connected with the corresponding control signal end of camera, three-dimensional moving stage, and described image acquiring method specifically comprises the following steps:
(1) three-dimensional moving stage is adopted to obtain preliminary panorama sketch: first, by computer control record three-dimensional moving stage surface level XY direction reference position, fiber cross section section is placed on three-dimensional moving stage, mobile Z-direction is to priori focusing position, adopt computer control that the motion track of three-dimensional moving stage in XY direction and moving step length are set, start objective table; Three-dimensional moving stage carries out work according to the aforementioned track that arranges, computer control starts camera and carries out fixed point shooting for the image under microscope at the moving step length pause point place of three-dimensional moving stage, after terminating, the image of fixing a point in moving process to take is sent to computer control by camera, computer control is spliced into the complete large figure of fiber cross section section, and three-dimensional moving stage moves to reference position;
(2) for preliminary panorama sketch and the large figure of complete fiber cross section section, computer control calculates fiber cross section and to cut into slices the projection of large figure in X, Y-direction, obtains the center of fiber cross section section;
(3) center is converted into objective table XY direction displacement, computer control drives three-dimensional moving stage that microscopical camera lens is moved to centre of slice position, the two benches search strategy based on priori focusing position is adopted to complete the auto-focusing of Z-direction, and repeat step (1), the fiber cross section section panorama sketch obtaining final focusing is again scanned in XY plane;
In step (1), described priori focusing position refers to the focusing position that the previous use of three-dimensional moving stage of computer control record produces afterwards;
In step (3), the auto-focusing adopting the two benches search strategy based on priori focusing position to complete Z-direction specifically comprises, first step search adopts larger objective table Z-direction step-length, adopts less step length searching when focusing operator size generation transition trend;
In step (3), find fiber cross section centre of slice position, moving stage, microscopical object lens center is made to aim at centre of slice position, objective table completes sectioning image auto-focusing, adopt gradient operator C as focusing operator in Z-direction one segment limit, carry out focus search, be best focusing position when C gets maximal value, operator as shown in the formula:
C ( z ) = Σ i = 1 M - 1 Σ j = 1 N - 1 ( ( I ( i + 1 , j ) - I ( i , j ) ) 2 + ( I ( i , j + 1 ) - I ( i , j ) ) 2 ) )
The span of z is the objective table Z-direction distance of swimming;
M is image X-direction number of pixels, and N is image Y-direction pixel number, I (i+1, j) the pixel gray scale I (i, j) that to be coordinate be (i+1, j) the pixel gray scale that to be coordinate be (i, j); I (i, j+1) the pixel gray scale that to be coordinate be (i, j+1).
2. fiber cross section section focusing digital picture acquisition methods under microscope according to claim 1, it is characterized in that in step (1), the moving step length of fixed point shooting is the lap of adjacent two width images, accounts for the 25-35% of picture traverse.
3. fiber cross section section focusing digital picture acquisition methods under microscope according to claim 1, it is characterized in that, in step (2), calculate the Gray Projection of X, Y-direction, when two direction projections are minimum value, position grouping is fiber cross section centre of slice position together.
CN201210257990.8A 2012-07-24 2012-07-24 Fiber cross section section focusing digital picture acquisition methods under a kind of microscope Expired - Fee Related CN102998787B (en)

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CN103728304A (en) * 2013-12-18 2014-04-16 宁波江丰生物信息技术有限公司 Focusing method for pathological section scanner
CN103760663A (en) * 2014-01-14 2014-04-30 宁波基内生物技术有限公司 Self-adaptive microscope, and use method and self-adaptive focusing method thereof
CN104390981B (en) * 2014-12-12 2017-07-14 宁波江丰生物信息技术有限公司 The method of quick and precisely focusing scanning pathological section tissue based on image collecting device
CN105181538B (en) * 2015-10-20 2018-06-26 丹东百特仪器有限公司 With scanning, the dynamic particle image granularity particle shape analyzer of splicing and method
CN105652429B (en) * 2016-03-22 2018-07-17 哈尔滨理工大学 A kind of microscopic cell glass slide scanning auto focusing method based on machine learning
CN106303242A (en) * 2016-08-18 2017-01-04 上海交通大学 The focusing system quickly of muti-spectrum imaging and method
CN106324795B (en) * 2016-09-29 2018-10-16 电子科技大学 A kind of automatic quick focusing method in more visuals field of detector microscope
CN108401446A (en) * 2017-08-24 2018-08-14 深圳市华显光学仪器有限公司 From moving axes microscope
CN109752831B (en) * 2017-11-02 2022-01-14 山东颐泽天泰医疗科技有限公司 Automatic focusing method, system and device for microscope with controllable focusing time
CN110727096B (en) * 2019-10-29 2022-03-18 北京临近空间飞行器系统工程研究所 Microscope focusing method and device and microscope equipment
CN113038003B (en) * 2021-02-26 2023-11-14 上海精测半导体技术有限公司 Automatic focusing device and method, and detection device and method

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CN102072935A (en) * 2010-10-26 2011-05-25 浙江大学 Automatic focusing method of scanning ultrasonic microscope
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CN102539295A (en) * 2012-02-16 2012-07-04 南京信息工程大学 Transgenic crop pollen real-time image automatic acquisition instrument and method

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