CN108320276A - Image taking alignment method and system - Google Patents
Image taking alignment method and system Download PDFInfo
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- CN108320276A CN108320276A CN201710024412.2A CN201710024412A CN108320276A CN 108320276 A CN108320276 A CN 108320276A CN 201710024412 A CN201710024412 A CN 201710024412A CN 108320276 A CN108320276 A CN 108320276A
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Abstract
The present invention relates to a kind of image taking alignment methods, including：An imaging device is provided, the imaging device includes the article carrying platform of the filming apparatus and face filming apparatus that obtain image, and article carrying platform can be moved along X-coordinate axis direction since reduction point or be moved along Y coordinate axis direction；Sample is placed on article carrying platform, movable object-carrying stage makes filming apparatus be observed sample, to determine a shot region in sample areas；Divide the subregion that the shot region is multiple arranged adjacents；The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out first time image taking, obtains the first image；The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out secondary image shooting, obtains the second image；And it calculates the offset between the first image and the second image and is scaled the displacement error of article carrying platform.Article carrying platform displacement error of the present invention is smaller.
Description
Technical field
The present invention relates to gene sequencing field, it is more particularly related to a kind of image taking alignment method and be
System.
Background technology
Gene sequencing field usually requires to carry out image taking and identification to sample, since the camera lens of filming apparatus observes model
The area much smaller than sample is enclosed, therefore camera lens every time can only shoot the part of sample.Prior art generally use is continuous
The mode of stepping shoots multiple zonules of the observation scope of adaptation imaging device successively, then spells the image of multiple zonules
It connects and is reduced to sample image.Due to needing repeatedly to be shot to each zonule of sample, the process that article carrying platform moves back and forth
The displacement error of middle generation prevents the image repeatedly shot from completely overlapped.
Invention content
The purpose of the present invention is to provide a kind of image taking alignment method and systems, it is intended to solve the bat of prior art image
When article carrying platform displacement error the problem of.
A kind of image taking alignment method, it is characterised in that including：
An imaging device is provided, the imaging device includes that the loading of the filming apparatus and face filming apparatus that obtain image is flat
Platform, article carrying platform can be moved along X-coordinate axis direction since reduction point and/or be moved along Y coordinate axis direction；
Sample is placed on article carrying platform, movable object-carrying stage makes filming apparatus be observed sample, to determine sample areas
An interior shot region；
Divide the subregion that the shot region is multiple arranged adjacents；
The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out first
Secondary image taking obtains the first image；
The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out secondary
Image taking obtains the second image；And
It calculates the offset between the first image and the second image and is scaled the displacement error of article carrying platform.
Preferably, described image shooting alignment method further comprises when the offset is more than default bias, by the
Two images recalculate the offset between the first image and the second image after the first image movement preset displacement, and convert
For the displacement error of article carrying platform.
Preferably, the preset displacement includes the displacement of the displacement and Y-direction of X-coordinate axis direction simultaneously.
Preferably, the displacement of the X-coordinate axis direction and the displacement of Y-direction are all higher than the default bias.
Preferably, described image shooting alignment method further comprises after the displacement error of dressing plate：Second condition
Lower movable object-carrying stage shoots image to the subregion in shot region.
Preferably, the first condition is to illuminate sample areas using light, and the second condition is that sample areas is spontaneous
Fluorescence.
Preferably, further comprise that movable object-carrying stage is to correct when the offset is in preset threshold range
The third image of filming apparatus shooting predetermined sub-region is controlled after offset and calculates the displacement error of article carrying platform again.
Preferably, movable object-carrying stage is to control the third image that filming apparatus shoots predetermined sub-region after correction offset
And the displacement error for calculating article carrying platform again includes：
Movable object-carrying stage and correction offset make filming apparatus align the predetermined sub-region under first condition and carry out third time
Image taking simultaneously obtains third image；
Calculate the offset between the first image and third image；And
If the offset between the first image and third image in preset threshold range, and the first image and third image it
Between offset be less than offset between the first image and the second image, then correcting captured device contraposition per sub-regions when carry
Otherwise the moving coordinate of object platform terminates the correction of article carrying platform displacement error.
A kind of image taking alignment system, it is characterised in that including：
Imaging device, the imaging device include the article carrying platform of the filming apparatus and face filming apparatus that obtain image, are carried
Object platform can be moved along X-coordinate axis direction since reduction point and/or be moved along Y coordinate axis direction；
Shot region determining module, the shot region determining module make filming apparatus observation bit for controlling movable object-carrying stage
In the sample on article carrying platform, and then determine a shot region in sample areas；
Shot region divides module, and the shot region segmentation module is multiple adjacent rows for controlling the segmentation shot region
Row subregion；
First image taking module makes filming apparatus pair for controlling the movable object-carrying stage since reduction point under first condition
The position predetermined sub-region simultaneously carries out first time image taking, obtains the first image；
Second image taking module makes filming apparatus pair for controlling the movable object-carrying stage since reduction point under first condition
The position predetermined sub-region simultaneously carries out secondary image shooting, obtains the second image；And
Displacement error computing module, for calculating the offset between the first image and the second image and being scaled article carrying platform
Displacement error.
Preferably, described image shooting alignment system further comprises calculating error correction module, when the offset is big
When default bias, calculates error correction module and the second image is recalculated first after the first image movement preset displacement
Offset between image and the second image.
Preferably, the preset displacement includes the displacement of the displacement and Y-direction of X-coordinate axis direction simultaneously.
Preferably, the displacement of the X-coordinate axis direction and the displacement of Y-direction are all higher than the default bias.
Preferably, the platform shift calibrating module further comprises moving coordinate correction module, is used for correcting captured dress
The moving coordinate of article carrying platform when setting contraposition per sub-regions.
Preferably, described image shooting alignment system further comprises common photo module, for moving under a second condition
Dynamic article carrying platform shoots image to the subregion in shot region.
Preferably, the first condition is to illuminate sample areas using light, and the second condition is that sample areas is spontaneous
Fluorescence.
Preferably, the platform shift calibrating module further comprises that secondary correction module, the secondary correction module are used
When judging to work as the offset in preset threshold range, movable object-carrying stage after correction offset to control filming apparatus
It shoots the third image of predetermined sub-region and calculates the displacement error of article carrying platform again.
Preferably, the secondary correction module further judges when the offset between the first image and third image is pre-
If in threshold range, and the offset between the first image and third image is less than the offset between the first image and the second image
When amount, the moving coordinate of article carrying platform when correcting captured device contraposition is per sub-regions.
Compared with the existing technology, image taking alignment method of the invention and system are to the formal shooting figure of multiple subregions
Before picture, first to predetermined sub-region（Contraposition refers to subregion）Image taking twice is carried out, and according to image taking result twice
The displacement error for correcting article carrying platform, can eliminate or reduce to greatest extent platform displacement error causes identical subregion is front and back to clap
Larger offset occurs between the multiple image taken the photograph, improves the flux of gene sequencing subsequent image data processing.Further, when
When the offset of image is more than default bias twice, it can also pass through the feature by the characteristic area of the second image with respect to the first image
Offset is recalculated after region movement preset displacement, correction in time calculates error, and the displacement error of article carrying platform is avoided to correct
It makes a fault, increases the stability of image taking alignment method and system of the present invention.In addition, the offset is in predetermined threshold value
When in range, it can also further shoot the third image of predetermined sub-region and calculate the displacement error of article carrying platform again, with
Correction calculates error in time, avoids the displacement error of article carrying platform from correcting and makes a fault, further increases image of the present invention and clap
Take the photograph the stability of alignment method and system.
Description of the drawings
Fig. 1 is the flow diagram of first embodiment of the invention image taking alignment method.
Fig. 2 is article carrying platform mobile route schematic diagram in Fig. 1 image pickup methods.
Fig. 3 is another mobile route schematic diagram of article carrying platform in Fig. 1 image pickup methods.
Fig. 4 is in Fig. 1 flows to the first image schematic diagram with characteristic area of predetermined sub-region shooting.
Fig. 5 is the schematic diagram of the second image with same characteristic features region to predetermined sub-region shooting in Fig. 1 flows.
Fig. 6 is the side-play amount schematic diagram between the first image and the second image shown in Fig. 4-5.
Fig. 7 is the secondary correction steps flow chart schematic diagram that Fig. 1 flows further comprise.
Fig. 8 is the side-play amount schematic diagram between the first image and third image in Fig. 7 secondary correction steps flow charts.
Fig. 9 is the calculating error correction step flow diagram that Fig. 1 flows further comprise.
Figure 10 is the schematic diagram that Fig. 9 calculates that characteristic area in error correction step moves preset displacement.
Figure 11 is the block diagram of an embodiment of the present invention image taking alignment system.
Specific implementation mode
In order to make the purpose , technical scheme and advantage of the present invention be clearer, below in conjunction with drawings and the embodiments,
The present invention will be described in further detail.
- Fig. 3 is please referred to Fig.1, first embodiment of the invention provides a kind of image taking alignment method comprising step
S11-S16。
Step S11, provides an imaging device, and the imaging device includes filming apparatus and the face bat for obtaining image
Take the photograph the article carrying platform 10 of device, article carrying platform 10 can be moved since reduction point o along X-coordinate axis direction and/or along Y-coordinate axle side
To movement.In present embodiment, filming apparatus includes camera lens and obtains the ccd image sensor of image.
Step S12 places sample 20 on article carrying platform 10, and movable object-carrying stage 10 makes filming apparatus carry out sample 20
Observation, to determine a shot region 30 in sample areas.In one embodiment, as shown in Fig. 2, the sample 20 is high-throughput base
Because sample is sequenced, the sample 20 includes multiple magnetic beads to be identified, is not easy to know due to magnetic bead small volume and in adjacent edges
Other situation, therefore the immediate vicinity rectangular area of selected high-throughput gene sequencing sample is as shot region 30, so as to accurate
Identify the information of 30 correspondence image of shot region.In this implementation, the reduction point is located at except shot region 30.Another embodiment party
In formula, as shown in figure 3, the sample 20 is high-throughput gene sequencing sample, the sample 20 includes multiple magnetic beads to be identified, by
There is in magnetic bead small volume and in adjacent edges situation not easy to identify, therefore the center of selected high-throughput gene sequencing sample is attached
Nearly bar-shaped zone is as shot region 30, to accurately identify the information of 30 correspondence image of shot region.It is described multiple in this implementation
Site is located at except shot region 30.
Step S13 divides the subregion 31 that the shot region 30 is multiple arranged adjacents.
In one embodiment, the focusing range of filming apparatus is much smaller than the area of standard specimen 20, the bat to gene sequencing sample
The preferable shot region 30 of imaging effect to be pre-selected is taken the photograph to carry out.When actual photographed, the shot region 30 that will choose in advance
It is divided into the subregion 31 of the focusing range of multiple adaptation imaging devices, then moves imaging device respectively to every sub-regions 31
It is shot.As shown in Fig. 2, in an embodiment, the subregion 31 is arranged in arrays and includes being arranged along X-coordinate axis direction
Multirow subregion A10-A50, B10-B50, C10-C50, D10-D50.As shown in figure 3, in another embodiment, the sub-district
Domain 31 is in a strip shape to be arranged and includes multiple subregion A10-A15, A20, A30-A35, the A40- arranged along Y coordinate axis direction
A44。
Step S14, the movable object-carrying stage since reduction point make under first condition filming apparatus align the pre- stator region
Domain simultaneously carries out first time image taking, obtains the first image.The subregion nearest apart from reduction point o defined in present embodiment
A10 is to align to refer to subregion, and in alternate embodiments, the contraposition according to circumstances can be selected arbitrarily with reference to subregion.
In one embodiment, referring to FIG. 2, from reduction point o along Y direction movable object-carrying stage 10, such as circuit in Fig. 2
Y1 makes the Y coordinate of filming apparatus and the first row subregion A10-A50（0, Y1）Alignment.Then along X-coordinate movable object-carrying stage
10, such as circuit X1 in Fig. 2, i.e., from the X-coordinate of reduction point（0, Y1）Start movable object-carrying stage 10, makes filming apparatus to being located at
The right margin subregion A10 of the first row subregion A10-A50 carries out first time shooting, to obtain the first image, this embodiment party
Right margin subregion A10 described in formula is the nearest subregion 31 of X-coordinate of the first row apart from reduction point as predetermined sub-region.Compared with
In good embodiment, the first condition is to illuminate sample areas using light.
In alternate embodiment, referring to FIG. 3, from reduction point o along X-direction movable object-carrying stage 10, such as circuit in Fig. 3
X1 makes the X-coordinate of filming apparatus and subregion A10（X1,0）Alignment, then along Y coordinate movable object-carrying stage 10, in Fig. 3
Circuit Y1 makes filming apparatus carry out first time shooting to being located at subregion A10, to obtain the first image.
Step S15, the movable object-carrying stage since reduction point make under first condition filming apparatus align the pre- stator region
Domain simultaneously carries out secondary image shooting, obtains the second image.
In one embodiment, referring to FIG. 2, again from reduction point o along Y direction movable object-carrying stage 10, such as Fig. 2 center lines
Road Y1 makes the Y coordinate of filming apparatus and the first row subregion A10-A50（0, Y1）Alignment.Then it is flat to move loading along X-coordinate
Platform 10, such as circuit X1 in Fig. 2, i.e., from the X-coordinate of reduction point（0, Y1）Start movable object-carrying stage 10, filming apparatus is made to align
Second is carried out in the right margin subregion A10 of the first row subregion A10-A50 to shoot, to obtain the second image.It is preferably real
It applies in mode, the first condition is to illuminate sample areas using light.
In alternate embodiment, referring to FIG. 3, again from reduction point o along X-direction movable object-carrying stage 10, such as Fig. 3 center lines
Road X1 makes the X-coordinate of filming apparatus and subregion A10（X1,0）Alignment, then along Y coordinate movable object-carrying stage 10, such as Fig. 3
Middle circuit Y1 makes filming apparatus carry out second of shooting to being located at subregion A10, to obtain the second image.
Step S16 calculates the offset between the first image and the second image and is scaled the displacement error of article carrying platform.
With reference to figure 4-5, the characteristic area 42 in the first image shown in Fig. 4 is calculated with respect to the first image corner 41（Also may be used
To be picture centre or other reference points）The first relative coordinate.Identify the same characteristic features region in the second image shown in fig. 5
52, and calculate same characteristic features region 52 and correspond to corner 51 with respect to the second image（Can also be picture centre or other reference points）'s
Second relative coordinate, the offset being then defined as the difference of the first, second relative coordinate between the first image and the second image
Offset, the displacement error of article carrying platform is scaled according to image and sample magnification ratio relationship k by amount.In present embodiment,
Characteristic area is to know the significantly identical standard specimen part of another characteristic from the first image and the second image by image-recognizing method,
In alternate embodiments, the characteristic area may be multiple.
In better embodiment, step S16 calculates the characteristic area 52 of the characteristic area 42 and the second image of the first image
Between offset still further comprise：The characteristic area 42 for defining the first image is m1, defines the characteristic area of the second image
52 be m2, and the calculating of the offset between m1 and m2 includes the following steps：
（1）M1 is converted into complex matrix m3, m2 is converted into complex matrix m4；
（2）Fast Fourier Transform (FFT) is asked respectively to m3 and m4；
（3）The conjugate matrices of m4 are sought, and energy spectrum is asked to m3 using this matrix；
（4）Inverse fast Fourier transform is asked to m3, time domain is turned again to from frequency domain；
（5）Matrix m is obtained to m3 real number matrix, matrix m is sorted, finds extreme point and its ranks coordinate, determine m1 and
The position of offset between m2 and offset.
In the present embodiment, if the characteristic area 52 of the characteristic area 42 of the first image and the second image there are displacement relation,
After Fourier transformation being carried out to them, the two Fourier transform amplitude having the same and different phases.The feature of first image
Phase relation after the Fourier transform of the characteristic area 52 of region 42 and the second image is determined by the displacement of the two, first
The phase difference of the characteristic area 42 of image and the characteristic area 52 of the second image is equal to the phase of its crosspower spectrum.
In alternate embodiments, it can also further comprise movable object-carrying stage pair under second condition after the step S16
Subregion in shot region shoots image.In present embodiment, article carrying platform moves simultaneously from reduction point o along X-axis, Y direction
Displacement error is corrected, filming apparatus is made to be aligned successively per sub-regions and carries out image taking.In one embodiment, if original every
The coordinate of sub-regions is（x,y）, the offset between the first image and the second image is illustrated in figure 6（w1,h1）, then correct
The coordinate of every sub-regions is after displacement error（x-kw1,y-kh1）.In the present embodiment, the second condition be sample areas from
It fluoresces.
Further include secondary correction step after step S16 in alternate embodiments with reference to figure 7, the secondary correction step
When judging that offset delta 1 is in preset threshold range, movable object-carrying stage is pre- to control filming apparatus shooting after correction offset
The third image of stator region and the displacement error for calculating article carrying platform again comprising step S171-S176.
In present embodiment, whether the step S171 calculates the offset delta 1 generated default for judgment step S16
In threshold range, if offset delta 1 is less than first threshold or is more than second threshold, terminates flow, stop article carrying platform correction
Displacement error.If offset delta 1 is in preset threshold range, i.e. second threshold >=1 >=first threshold of offset delta then executes step
Rapid S172.
In present embodiment, the step S172 is used to control movable object-carrying stage with correction offset δ 1, such as will carry
Object platform is moved to the coordinate of predetermined sub-region after correction, in an embodiment, if the former coordinate of predetermined sub-region is（x0,
y0）, the offset between the first image and the second image is illustrated in figure 6（w1,h1）, then pre- stator region after correction offset δ 1
The coordinate in domain is（x0-kw1,y0-kh1）, so that filming apparatus is aligned the predetermined sub-region and carry out third time figure
As shooting, third image is obtained.
In present embodiment, the step S173 is for calculating the offset delta 2 between the first image and third image and changing
Calculate the displacement error for article carrying platform.In one embodiment, with reference to the method identical with step S16 of figure 8, identify in third image
Characteristic area 62, and calculate 62 third relative coordinate of characteristic area, be then defined as the difference of first, third relative coordinate
Offset is scaled loading by the offset between the first image and third image according to image and sample magnification ratio relationship k
The displacement error of platform.In present embodiment, characteristic area be by image-recognizing method from the first image and third image
Know the significantly identical standard specimen part of another characteristic.In one specific implementation mode, the changing coordinates of predetermined sub-region 62 are（x0-kw1,
y0-kh1）, the offset delta 2 between the first image and third image is illustrated in figure 8（w2,h2）, then correct pre- after displacement error
The coordinate of stator region is（x0-kw2-kw1,y0-kh2-kh1）.
In present embodiment, whether the step S174 calculates the offset delta 2 generated default for judgment step S173
In threshold range, if offset delta 2 is less than first threshold or is more than second threshold, terminates flow, stop article carrying platform correction
Displacement error.If offset delta 2 is in preset threshold range, i.e. second threshold >=1 >=first threshold of offset delta, step is executed
S176。
In present embodiment, the step S175 judges whether offset delta 2 is less than offset delta 1 for further, if
Offset delta 2 is less than offset delta 1, thens follow the steps S176, and each sub-district is aligned according to offset delta 1 and 2 correcting captured devices of δ
The moving coordinate of article carrying platform when domain.If offset delta 2 is more than or equal to offset delta 1, terminate flow, stops or terminate loading
Stage correction displacement error.
In present embodiment, the offset is judged using absolute value, and the first threshold can be 6-10 picture
Plain distance, second threshold can be 700-900 pixel distance, in better embodiment, the first threshold for 8 pixels away from
From second threshold can be 800 pixel distances, and the preset threshold range is 8-800 pixel distances.
With reference to figure 9, in other alternate embodiments, further includes step after step S16, specifically include step S91-S94.
Step S91 executes step S92 when judging that the offset delta 1 is more than or equal to default bias, no to then follow the steps
S94。
Second image is recalculated the first image and the second figure by step S92 after the first image movement preset displacement
Offset as between.In preferred embodiment, the characteristic area by the characteristic area of the second image relative to the first image moves in advance
If recalculating the offset between the first image and the second image after displacement, and it is scaled the displacement error of article carrying platform.
Step S93 terminates flow and stops correction, otherwise hold when judging that the offset delta 2 is more than or equal to default bias
Row step S94, the moving coordinate of article carrying platform when being aligned per sub-regions with 2 correcting captured device of offset delta.
In the present embodiment, the preset displacement includes the displacement of the displacement and Y-direction of X-coordinate axis direction simultaneously, and
The preset displacement is all higher than the default default bias in the displacement of X-coordinate axis direction and the displacement of Y-direction, such as greatly
In 2 pixel distances.In one embodiment, as shown in Figure 10, when the offset delta 1 between the first image and the second image is in X-axis side
It is 0 in Y direction to for W1, and when W1 is more than default bias, by the characteristic area 52 of the second image along X-direction is tilted, moves
Dynamic preset displacement（W0,W0）Afterwards, the offset delta 2 between the first image and the second image is recalculated.
Specifically it is calculated as：The characteristic area 42 for defining the first image is m1, and the characteristic area 52 for defining the second image moves
Dynamic preset displacement（W0,W0）Characteristic area 52 ' afterwards is m2, and the calculating of the correction offset between m1 and m2 includes the following steps：
（1）M1 is converted into complex matrix m3, m2 is converted into complex matrix m4；
（2）Fast Fourier Transform (FFT) is asked respectively to m3 and m4；
（3）The conjugate matrices of m4 are sought, and energy spectrum is asked to m3 using this matrix；
（4）Inverse fast Fourier transform is asked to m3, time domain is turned again to from frequency domain；
（5）Matrix m is obtained to m3 real number matrix, matrix m is sorted, finds extreme point and its ranks coordinate, determine m1 and
The position of offset between m2 and correction offset.The position of offset between m1 and m2 and correction offset subtract mobile default
Displacement（W0,W0）The offset delta 2 between the first image and the second image is obtained afterwards.
In present embodiment, the generally ramped X-direction 5-95 degree in direction of the movement of characteristic area 52 ' of the second image, compared with
In good embodiment, the mobile direction of characteristic area 52 ' tilts 30 degree, 45 degree or 60 degree of X-axis.In Figure 10 the embodiment described,
Characteristic area 52 ' tilts 45 degree of X-axis from the direction that characteristic area 52 moves.
1 is please referred to Fig.1, third embodiment of the invention provides a kind of image taking alignment system 100, described image shooting
Alignment system 100 includes imaging device 110, shot region determining module 120, shot region segmentation module 130, the bat of the first image
Take the photograph module 140, the second image taking module 150, displacement error computing module 160, moving coordinate correction module 170, secondary school
Positive module 180 calculates error correction module 185 and common photo module 190.In present embodiment, the module or system can
Can also be that storage executes completion specific function in memory and by processor to be the integrated circuit for executing specific function
Software program.
Please also refer to Fig. 2, the imaging device 110 includes the filming apparatus and face filming apparatus for obtaining image
Article carrying platform 10, article carrying platform 10 can be moved along X-coordinate axis direction since reduction point o and/or be moved along Y coordinate axis direction.
In present embodiment, filming apparatus includes camera lens and obtains the ccd image sensor of image.
The shot region determining module 120 keeps filming apparatus observation flat positioned at loading for controlling movable object-carrying stage 10
Sample 20 on platform 10, and then determine a shot region 30 in sample areas.In one embodiment, as shown in Fig. 2, the sample
20 be high-throughput gene sequencing sample, and the sample 20 includes multiple magnetic beads to be identified, due to magnetic bead small volume and at edge
Nearby there is situation not easy to identify, therefore the immediate vicinity rectangular area of selected high-throughput gene sequencing sample is as shot region
30, to accurately identify the information of 30 correspondence image of shot region.In this implementation, the reduction point be located at shot region 30 it
Outside.In another embodiment, as shown in figure 3, the sample 20 is high-throughput gene sequencing sample, the sample 20 includes multiple
Magnetic bead to be identified has situation not easy to identify due to magnetic bead small volume and in adjacent edges, selectes high throughput gene and surveys
The immediate vicinity bar-shaped zone of sequence sample is as shot region 30, to accurately identify the information of 30 correspondence image of shot region.
In this implementation, the reduction point is located at except shot region 30.
The shot region segmentation module 130 is used to control the sub-district for dividing that the shot region 30 is multiple arranged adjacents
Domain 31.In one embodiment, the focusing range of filming apparatus is much smaller than the area of standard specimen 20, to the shooting of gene sequencing sample with
The preferable shot region 30 of imaging effect being pre-selected carries out.When actual photographed, the shot region 30 chosen in advance is divided
For the subregion 31 of the focusing range of multiple adaptation imaging devices, then moves imaging device and every sub-regions 31 are carried out respectively
Shooting.As shown in Fig. 2, in an embodiment, the subregion 31 it is arranged in arrays and include arranged along X-coordinate axis direction it is more
Row subregion.As shown in figure 3, in another embodiment, the subregion 31 is in a strip shape to be arranged and includes being arranged along Y coordinate axis direction
Multiple subregions of row.
Described first image taking module 140 makes for controlling the movable object-carrying stage since reduction point under first condition
Filming apparatus aligns the predetermined sub-region and carries out first time image taking, obtains the first image.
In one embodiment, referring to FIG. 2, from reduction point o along Y direction movable object-carrying stage 10, such as circuit in Fig. 2
Y1 makes the Y coordinate of filming apparatus and the first row subregion A10-A50（0, Y1）Alignment.Then along X-coordinate movable object-carrying stage
10, such as circuit X1 in Fig. 2, i.e., from the X-coordinate of reduction point（0, Y1）Start movable object-carrying stage 10, makes filming apparatus to being located at
The right margin subregion A10 of the first row subregion A10-A50 carries out first time shooting, to obtain the first image, this embodiment party
Right margin subregion A10 described in formula is the nearest subregion 31 of X-coordinate of the first row apart from reduction point as predetermined sub-region.Compared with
In good embodiment, the first condition is to illuminate sample areas using light.In alternate embodiment, referring to FIG. 3, from reset
Point o makes the X-coordinate of filming apparatus and subregion A10 along X-direction movable object-carrying stage 10 such as circuit X1 in Fig. 3（X1,0）
Then alignment makes filming apparatus be carried out to being located at subregion A10 along Y coordinate movable object-carrying stage 10 such as circuit Y1 in Fig. 3
It shoots for the first time, to obtain the first image.
The second image taking module 150 makes for controlling the movable object-carrying stage since reduction point under first condition
Filming apparatus aligns the predetermined sub-region and carries out second of image taking, obtains the second image.
As shown in Fig. 2, in an embodiment, again from reduction point o along Y direction movable object-carrying stage 10, such as circuit in Fig. 2
Y1 makes the Y coordinate of filming apparatus and the first row subregion A10-A50（0, Y1）Alignment.Then along X-coordinate movable object-carrying stage
10, such as circuit X1 in Fig. 2, i.e., from the X-coordinate of reduction point（0, Y1）Start movable object-carrying stage 10, makes filming apparatus to being located at
The right margin subregion A10 of the first row subregion A10-A50 carries out second and shoots, to obtain the second image.Preferably implement
In mode, the first condition is to illuminate sample areas using light.As shown in figure 3, in an alternate embodiment, again from reset
Point o makes the X-coordinate of filming apparatus and subregion A10 along X-direction movable object-carrying stage 10 such as circuit X1 in Fig. 3（X1,0）
Then alignment makes filming apparatus be carried out to being located at subregion A10 along Y coordinate movable object-carrying stage 10 such as circuit Y1 in Fig. 3
Second of shooting, to obtain the second image.
The displacement error computing module 160 is for calculating the offset between the first image and the second image and being scaled
The displacement error of article carrying platform.With reference to figure 4-5, in preferred embodiment, the characteristic area 42 in the first image shown in Fig. 4 is calculated
Opposite first image corner 41（Can also be picture centre or other reference points）The first relative coordinate.It identifies shown in fig. 5
Same characteristic features region 52 in second image, and calculate same characteristic features region 52 and correspond to corner 51 with respect to the second image（It can also
It is picture centre or other reference points）The second relative coordinate, the difference of the first, second relative coordinate is then defined as first
Offset is scaled article carrying platform by the offset between image and the second image according to image and sample magnification ratio relationship k
Displacement error.In present embodiment, characteristic area is to be identified from the first image and the second image by image-recognizing method
The significantly identical standard specimen part of feature, in alternate embodiments, the characteristic area may be multiple.
It is inclined between the characteristic area 42 and the characteristic area 52 of the second image of the first image of calculating in better embodiment
Shifting amount still further comprises：The characteristic area 42 for defining the first image is m1, and the characteristic area 52 for defining the second image is m2, m1
The calculating of offset between m2 includes the following steps：
（1）M1 is converted into complex matrix m3, m2 is converted into complex matrix m4；
（2）Fast Fourier Transform (FFT) is asked respectively to m3 and m4；
（3）The conjugate matrices of m4 are sought, and energy spectrum is asked to m3 using this matrix；
（4）Inverse fast Fourier transform is asked to m3, time domain is turned again to from frequency domain；
（5）Matrix m is obtained to m3 real number matrix, matrix m is sorted, finds extreme point and its ranks coordinate, determine m1 and
The position of offset between m2 and offset.
In the present embodiment, if the characteristic area 52 of the characteristic area 42 of the first image and the second image there are displacement relation,
After Fourier transformation being carried out to them, the two Fourier transform amplitude having the same and different phases.The feature of first image
Phase relation after the Fourier transform of the characteristic area 52 of region 42 and the second image is determined by the displacement of the two, first
The phase difference of the characteristic area 42 of image and the characteristic area 52 of the second image is equal to the phase of its crosspower spectrum.
Article carrying platform when the moving coordinate correction module 170 is used to control correcting captured device contraposition per sub-regions
Moving coordinate.In present embodiment, article carrying platform moves along X-axis, Y direction from reduction point o and corrects displacement error, makes shooting
Device aligns successively per sub-regions and carries out image taking.In one embodiment, if the original coordinate per sub-regions is（x,
y）, the offset between the first image and the second image is illustrated in figure 6（w,h）, then every sub-regions after displacement error are corrected
Coordinate be（x-kw,y-kh）.
The secondary correction module 180 is for judging when the offset is in preset threshold range, control shooting
Device shoots the third image of predetermined sub-region and calculates the displacement error of article carrying platform again.When the first image and third image
Between offset in preset threshold range, and the offset between the first image and third image is less than the first image and the
When offset between two images, the moving coordinate of article carrying platform when correcting captured device contraposition is per sub-regions.
In one specific implementation mode, whether the secondary correction module 180 judges the offset delta 1 generated beyond default threshold
It is worth range, if exceeding preset threshold range, the secondary correction module 180 terminates flow, stops article carrying platform movement and misses
Difference correction.If in preset threshold range, movable object-carrying stage makes shooting fill with correction offset δ 1 again under first condition
It sets the contraposition predetermined sub-region and carries out third time image taking, obtain third image.Then the first image and third are calculated
Offset delta 2 between image；Then judge whether offset delta 2 exceeds preset threshold range, if exceeding preset threshold range,
Then the secondary correction module 180 terminates flow, stops the correction of article carrying platform displacement error.If offset delta 2 is in predetermined threshold value
In range, further judge whether offset delta 2 is less than offset delta 1, if offset delta 2 is less than offset delta 1, according to offset
The moving coordinate of article carrying platform when measuring δ 1 and the contraposition of 2 correcting captured devices of δ per sub-regions.If offset delta 2 is more than or equal to inclined
Shifting amount δ 1 then terminates flow, stops article carrying platform and corrects displacement error.In present embodiment, the preset threshold range is 8-
800 pixel distances.In alternate embodiments, the detailed description about secondary correction module 180 is referring also to abovementioned steps
S171-S176。
When the calculating error correction module 185 judges that the offset delta 1 is more than or equal to default bias, by the second image
The offset between the first image and the second image is recalculated after the first image movement preset displacement.Preferred embodiment
In, by the characteristic area of the second image relative to the first image characteristic area movement preset displacement after recalculate the first image and
Offset delta 2 between second image, and it is scaled the displacement error of article carrying platform.
In the present embodiment, the preset displacement includes the displacement of the displacement and Y-direction of X-coordinate axis direction simultaneously, and
The preset displacement is all higher than the default default bias in the displacement of X-coordinate axis direction and the displacement of Y-direction.One implements
In example, as shown in Figure 10, the characteristic area 42 for defining the first image is m1, and the movement of characteristic area 52 for defining the second image is default
Displacement（W0,W0）Characteristic area 52 ' afterwards is m2, and the calculating of the correction offset between m1 and m2 includes the following steps：
（1）M1 is converted into complex matrix m3, m2 is converted into complex matrix m4；
（2）Fast Fourier Transform (FFT) is asked respectively to m3 and m4；
（3）The conjugate matrices of m4 are sought, and energy spectrum is asked to m3 using this matrix；
（4）Inverse fast Fourier transform is asked to m3, time domain is turned again to from frequency domain；
（5）Matrix m is obtained to m3 real number matrix, matrix m is sorted, finds extreme point and its ranks coordinate, determine m1 and
The position of offset between m2 and correction offset.The position of offset between m1 and m2 and correction offset subtract mobile default
Displacement（W0,W0）The offset delta 2 between the first image and the second image is obtained afterwards.
In present embodiment, the generally ramped X-direction 5-95 degree in direction of the movement of characteristic area 52 ' of the second image, compared with
In good embodiment, the mobile direction of characteristic area 52 ' tilts 30 degree, 45 degree or 60 degree of X-axis.In Figure 10 the embodiment described,
Characteristic area 52 ' tilts 45 degree of X-axis from the direction that characteristic area 52 moves.
After the displacement error of dressing plate, the common photo module 190 is for controlling stepping movable object-carrying stage successively
Image taking under a second condition is carried out to every sub-regions, until the image taking of entire shot region.Present embodiment
In, the second condition is sample areas autofluorescence.
Compared with the existing technology, image taking alignment method of the invention and system are to the formal shooting figure of multiple subregions
Before picture, first to predetermined sub-region（Contraposition refers to subregion）Image taking twice is carried out, and according to image taking result twice
The displacement error for correcting article carrying platform, can eliminate or reduce to greatest extent platform displacement error causes identical subregion is front and back to clap
Larger offset occurs between the multiple image taken the photograph, improves the flux of gene sequencing subsequent image data processing.Further, when
When the offset of image is more than default bias twice, it can also pass through the feature by the characteristic area of the second image with respect to the first image
Offset is recalculated after region movement preset displacement, correction in time calculates error, and the displacement error of article carrying platform is avoided to correct
It makes a fault, increases the stability of image taking alignment method and system of the present invention.In addition, the offset is in predetermined threshold value
When in range, it can also further shoot the third image of predetermined sub-region and calculate the displacement error of article carrying platform again, with
Correction calculates error in time, avoids the displacement error of article carrying platform from correcting and makes a fault, further increases image of the present invention and clap
Take the photograph the stability of alignment method and system.
The foregoing is merely the better embodiments of the present invention, are not intended to limit the invention, all the present invention's
All any modification, equivalent and improvement etc., should all be included in the protection scope of the present invention made by within spirit and principle.
Claims (17)
1. a kind of image taking alignment method, it is characterised in that including：
An imaging device is provided, the imaging device includes that the loading of the filming apparatus and face filming apparatus that obtain image is flat
Platform, article carrying platform can be moved along X-coordinate axis direction since reduction point and/or be moved along Y coordinate axis direction；
Sample is placed on article carrying platform, movable object-carrying stage makes filming apparatus be observed sample, to determine sample areas
An interior shot region；
Divide the subregion that the shot region is multiple arranged adjacents；
The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out first
Secondary image taking obtains the first image；
The movable object-carrying stage since reduction point makes under first condition filming apparatus align the predetermined sub-region and carries out secondary
Image taking obtains the second image；And
It calculates the offset between the first image and the second image and is scaled the displacement error of article carrying platform.
2. image taking alignment method according to claim 1, which is characterized in that further comprise when the offset is big
When default bias, by the second image relative to the first image move preset displacement after recalculate the first image and the second image it
Between offset, and be scaled the displacement error of article carrying platform.
3. image taking alignment method according to claim 2, which is characterized in that the preset displacement is sat including X simultaneously
The displacement in parameter direction and the displacement of Y-direction.
4. image taking alignment method according to claim 3, which is characterized in that the displacement of the X-coordinate axis direction
It is all higher than the default bias with the displacement of Y-direction.
5. image taking alignment method according to claim 1, which is characterized in that the displacement error laggard one of dressing plate
Step includes：Movable object-carrying stage shoots image to the subregion in shot region under second condition.
6. image taking alignment method according to claim 5, which is characterized in that the first condition is to be shone using light
Bright sample areas, the second condition are sample areas autofluorescence.
7. image taking alignment method according to claim 1, which is characterized in that further comprise existing when the offset
When in preset threshold range, movable object-carrying stage is to control the third that filming apparatus shoots predetermined sub-region after correction offset
Image and the displacement error for calculating article carrying platform again.
8. image taking alignment method according to claim 7, which is characterized in that movable object-carrying stage is with correction offset
Control filming apparatus shoots the third image of predetermined sub-region afterwards and the displacement error of calculating article carrying platform includes again：
Movable object-carrying stage and correction offset make filming apparatus align the predetermined sub-region under first condition and carry out third time
Image taking simultaneously obtains third image；
Calculate the offset between the first image and third image；And
If the offset between the first image and third image in preset threshold range, and the first image and third image it
Between offset be less than offset between the first image and the second image, then correcting captured device contraposition per sub-regions when carry
Otherwise the moving coordinate of object platform terminates the correction of article carrying platform displacement error.
9. a kind of image taking alignment system, it is characterised in that including：
Imaging device, the imaging device include the article carrying platform of the filming apparatus and face filming apparatus that obtain image, are carried
Object platform can be moved along X-coordinate axis direction since reduction point and/or be moved along Y coordinate axis direction；
Shot region determining module, the shot region determining module make filming apparatus observation bit for controlling movable object-carrying stage
In the sample on article carrying platform, and then determine a shot region in sample areas；
Shot region divides module, and the shot region segmentation module is multiple adjacent rows for controlling the segmentation shot region
Row subregion；
First image taking module makes filming apparatus pair for controlling the movable object-carrying stage since reduction point under first condition
The position predetermined sub-region simultaneously carries out first time image taking, obtains the first image；
Second image taking module makes filming apparatus pair for controlling the movable object-carrying stage since reduction point under first condition
The position predetermined sub-region simultaneously carries out secondary image shooting, obtains the second image；And
Displacement error computing module, for calculating the offset between the first image and the second image and being scaled article carrying platform
Displacement error.
10. image taking alignment system according to claim 9, which is characterized in that further comprise calculating error correction
Module calculates error correction module and moves the second image in advance relative to the first image when the offset is more than default bias
If recalculating the offset between the first image and the second image after displacement.
11. image taking alignment system according to claim 10, which is characterized in that the preset displacement includes X simultaneously
The displacement of change in coordinate axis direction and the displacement of Y-direction.
12. image taking alignment system according to claim 11, which is characterized in that the displacement of the X-coordinate axis direction
The displacement of amount and Y-direction is all higher than the default bias.
13. image taking alignment system according to claim 9, which is characterized in that the platform shift calibrating module into
One step includes moving coordinate correction module, the moving coordinate of article carrying platform when for the contraposition of correcting captured device per sub-regions.
14. image taking alignment system according to claim 9, further comprises common photo module, for second
Under the conditions of movable object-carrying stage in shot region subregion shoot image.
15. image taking alignment system according to claim 14, which is characterized in that the first condition is to use light
Sample areas is illuminated, the second condition is sample areas autofluorescence.
16. image taking alignment system according to claim 9, which is characterized in that the platform shift calibrating module into
One step includes secondary correction module, and the secondary correction module is for judging when the offset is in preset threshold range
It waits, movable object-carrying stage is carried with controlling the third image of filming apparatus shooting predetermined sub-region after correction offset and calculating again
The displacement error of object platform.
17. image taking alignment system according to claim 16, which is characterized in that the secondary correction module is further
Judge to work as the offset between the first image and third image in preset threshold range, and between the first image and third image
Offset when being less than offset between the first image and the second image, loading when correcting captured device contraposition is per sub-regions
The moving coordinate of platform.
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