CN104516098B - Microscopy device and imaging method - Google Patents
Microscopy device and imaging method Download PDFInfo
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- CN104516098B CN104516098B CN201310461837.1A CN201310461837A CN104516098B CN 104516098 B CN104516098 B CN 104516098B CN 201310461837 A CN201310461837 A CN 201310461837A CN 104516098 B CN104516098 B CN 104516098B
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/365—Control or image processing arrangements for digital or video microscopes
- G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
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- G—PHYSICS
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Abstract
The invention provides a microscopy device and an imaging method. The device comprises a displacement device, an imaging device, a position sensing device and an image processing device. The displacement device is used for allowing a relative displacement to be generated between a to-be-detected sample and the imaging device, and the displacement is located in the same plane. The imaging device is used for acquiring multiple images of the to-be-detected sample relative to the imaging device and at different positions, and the multiple images have first resolution. The position sensing device is used for detecting the relative displacement between the to-be-detected sample and the imaging device. The image processing device is sued for receiving the multiple images with the first resolution, receiving the displacement detected by the position sensing device, and overlaying the multiple images according to the displacement to synthesize images with second resolution higher than the first resolution. By the microscopy device and the imaging method, high-magnification lenses are not needed, and high-resolution images can be obtained in a large view field.
Description
Technical field
The present invention relates to a kind of microscope equipment and its imaging method, more particularly to one kind by being superimposed several sampled images and
Form the microscope equipment and its imaging method of final micro-image.
Background technology
In microscope equipment, in order to improve the resolution ratio of image, the high amplification with large-numerical aperture (NA) is usually used
The lens of multiple are obtaining image.However, the visual field (field of view, FOV) of the lens of this high-amplification-factor is put than low
The visual field of the lens of big multiple is less, therefore, this microscope equipment generally utilizes motor mobile example platform, to obtain sample not
With the image of the high-amplification-factor at position, recycle software that these image mosaics are got up, form high-amplification-factor, high-resolution
Rate image.As depicted in figs. 1 and 2, wherein Fig. 1 is the side view of microscope equipment to the configuration example of this microscope equipment, and Fig. 2 is micro-
The top view of device.As depicted in figs. 1 and 2, microscope equipment of the prior art includes imageing sensor 1, camera lens 2, sample stage
4th, control device 5, light source 6 and motor 7.When needing to obtain high-resolution image, the lens of high magnification numbe are first selected, then using horse
Up to 7 mobile example platforms, to obtain the image of the high-amplification-factor of the different parts of sample, after splicing formed high-amplification-factor,
High-resolution image.One example of this microscope equipment is, for example, being used for disclosed in United States Patent (USP) US2009/0168160
The microscopic system of scanning cell sample, it passes through the sample stage mobile example of mechanization and sample is repositioned.
But it is this to be wasted time and energy using the method for being spliced to form high-definition picture, it is not easy to application.Therefore, develop
A kind of new technology, it forms final micro-image by being superimposed several sampled images.Such as United States Patent (USP) US2012/
Without lens microscope equipment, its basic structure is as shown in figure 3, including sample cell 04, bag for scanning, projection disclosed in 0098950
Include the imageing sensor 01 and processor 05 on the outside of the scanning light source of multiple light-emitting components 06, sample surfaces.The microscope equipment
By multiple light-emitting components 06 blood sample in sample cell 04 is irradiated successively, respectively obtains the figure of multiple low resolution
Picture, because the image of each low resolution is formed by different light-emitting component irradiations, and the photograph of different light-emitting components
Firing angle degree is again different, therefore has certain skew (having relative displacement between image) between the image of each low resolution.
Then, according to the position relationship between multiple light-emitting components, the image of multiple low resolution can be synthesized a panel height resolution ratio
Image.This method for forming final micro-image by being superimposed several sampled images can obtain the high-resolution of big visual field
Image, without the need for splicing, it is more quick and be easy to apply.But, due to the limited area of the light-emitting component in scanning light source, its
The light for going out not directional light, causes same light source different to the irradiating angle of each position of sample, therefore each position of sample
There is difference in the projective distribution put, easily the periphery in sample goes out to produce pattern distortion.
The content of the invention
The present invention is intended to provide a kind of microscope equipment and imaging method, can obtain the full resolution pricture of big visual field, while not
Pattern distortion can be produced.
The invention provides a kind of microscope equipment, including：
Gearshift, for making to produce relative displacement between testing sample and imaging device, institute's bit shifts are flat in one
In face；
Imaging device, for obtaining several figures of the testing sample in the various location relative to the imaging device
Picture, wherein the multiple image has first resolution；
Position sensing apparatus, for detecting the relative displacement between the testing sample and the imaging device；
Image processing apparatus, for receiving the multiple image with first resolution and position sensing apparatus inspection
The displacement for measuring, and described several are had by the imaging importing of first resolution according to the displacement for detecting, synthesis
Go out the image with the second resolution higher than first resolution.The microscope equipment can be regarded greatly without the need for the lens of high magnification numbe
The full resolution pricture of field.
According to the microscope equipment that the present invention is provided, wherein the gearshift is used to be subjected to displacement the testing sample.
According to the microscope equipment that the present invention is provided, wherein the gearshift is used to be subjected to displacement the imaging device.
According to the microscope equipment that the present invention is provided, also include the light for producing the directional light of the irradiation testing sample
Source, can avoid the generation of pattern distortion.
According to the microscope equipment that the present invention is provided, including for sending the pulsed light of the light for irradiating the testing sample
Source.
According to the microscope equipment that the present invention is provided, wherein the plane is horizontal plane.
According to the microscope equipment that the present invention is provided, wherein the gearshift includes stopping means.
According to the microscope equipment that the present invention is provided, wherein at interval of a sampling interval duration, the imaging device obtains one
The image of testing sample described in width.
According to the microscope equipment that the present invention is provided, wherein the sampling interval duration is selected as making to be adopted described in one, interval
Displacement between the adjacent image with first resolution of sample interval time is equal to of the imaging device
The 1/10-1/3 of the size of pixel.
According to the present invention provide microscope equipment, wherein the displacement amplitude of the gearshift be chosen to it is described many
Width has the maximum relative displacement amount between the image of first resolution more than the size of a pixel of the imaging device.
According to the microscope equipment that the present invention is provided, wherein the gearshift is vibrating device.
According to the microscope equipment that the present invention is provided, wherein the microscope equipment is transmission-type or reflecting microscope, or
Projection is without lens microscope equipment.
According to the microscope equipment that the present invention is provided, wherein the position sensing apparatus include the reference mark on testing sample
Note, reference marker imaging on the imaging device covers multiple pixels of the imaging device, according to described many
Relativeness between the gray value of each pixel in individual pixel determines position of the reference marker relative to the imaging device.
The present invention also provides a kind of imaging method of microscope equipment, including：
Obtain testing sample using imaging device and multiple image with first resolution；
The testing sample is set to form relative displacement with the imaging device, institute's bit shifts are in a plane；
Detect the relative shift between the testing sample and the imaging device；
Several images with first resolution and detected displacement are received, and according to detected displacement
By the superposition of the multiple image with first resolution, synthesize the figure with the second resolution higher than first resolution
Picture.
Microscope equipment and imaging method that the present invention is provided, without the need for the lens of high magnification numbe, in big visual field height are also obtained in that
The image of resolution.Further, since light source need not adopt multiple light sources, therefore the larger parallel light source of area can be adopted, therefore
Pattern distortion will not be produced.
Description of the drawings
The following drawings is only intended in schematic illustration and explanation is done to the present invention, is not delimit the scope of the invention.Wherein,
Fig. 1 is the side view of microscope equipment of the prior art；
Fig. 2 is the top view of microscope equipment of the prior art；
Fig. 3 is structural representation of the projection of the prior art without lens microscope equipment；
Fig. 4 is the side view of the microscope equipment according to embodiments of the invention 1；
Fig. 5 is the top view of the microscope equipment according to embodiments of the invention 1；
Fig. 6 is the side view of the microscope equipment according to embodiments of the invention 2；
Fig. 7 is side view of the projection without lens microscope equipment according to embodiments of the invention 3；
Fig. 8 a show the method for high-definition picture is obtained by stitching image in prior art；
Fig. 8 b show the method for obtaining high-definition picture employed in the present invention；
Fig. 9 shows the simulation result of microscope equipment of the invention and imaging method；
Figure 10 shows the another simulation result of microscope equipment of the invention and imaging method；
Figure 11 shows a kind of location measurement method of the invention；
Figure 12 shows another location measurement method of the invention.
Specific embodiment
In order to be more clearly understood to the technical characteristic of the present invention, purpose and effect, now control illustrates this
Bright specific embodiment.
Microscope equipment and imaging method that the present invention is provided, by displacement modes such as vibrations between sample and imaging device
Form relative displacement.Imaging device obtains the image of a width sample, and we are referred to as the image of this imaging device direct access
" low-resolution image ".Due to forming relative displacement between sample and imaging device so that have between multiple low-resolution images
There is certain displacement.In addition, sample is filled with imaging when obtaining each width low-resolution image using position sensing apparatus record
Relative shift between putting such that it is able to obtain the relative shift between low-resolution image.Over time, according to
The all low-resolution images superposition obtained in the time period is synthesized a high-resolution image by the relative shift.
Microscope equipment and imaging method that the present invention is provided, without the need for the lens of high magnification numbe, can also obtain full resolution pricture under big visual field,
And will not produce pattern distortion.
Embodiment 1
The present embodiment provides a kind of microscope equipment, and as shown in Figure 4 and Figure 5, wherein Fig. 4 is the side-looking of microscope equipment to its structure
Figure, Fig. 5 is the top view of microscope equipment.The microscope equipment includes：
Light source 16, the light for being sent can be directional light；
Sample stage 14, for carrying sample 13；
Imaging device, including lens 12 and imageing sensor 11, for obtaining the figure of the sample 13 in the case where light source 16 irradiates
Picture；
Vibrating device, including 8 springs 15 and vibrating motor 17, wherein vibrating motor 17 are used to vibrate sample stage 14,
So as to produce relative displacement between the sample 13 that carries on the sample stage 14 and imaging device, 8 springs 15 are respectively fixed to sample
Four sides of sample platform, for the plane internal vibration for making sample stage 14 only be located in sample stage 14；
Position sensor 18, the relative shift of the sample stage 14 when vibrating for real-time detection；
Image processing apparatus 19, for controlling light source 16 pulsed light is sent, and controls many of imaging device acquisition sample 13
Width low-resolution image, and record the phase of the sample stage 14 that position sensor 18 is detected when obtaining every width low-resolution image
To displacement, then the low-resolution image for obtaining is superimposed according to the relative shift of sample stage 14, synthesizes high-resolution
Image.
In the microscope equipment that the present embodiment is provided, light source 16, sample stage 14 and imaging device and the prior art for being adopted
In microscope equipment structure it is identical, need obtain low-resolution image when, can be as microscope equipment of the prior art
Directly the sample on sample stage 14 is imaged using imaging device.When needing to obtain high-definition picture, the present embodiment is provided
The imaging method of microscope equipment include：
The vibration frequency of vibrating motor 17 is set, sample stage 14 is vibrated, so as to the sample 13 carried on sample stage 14
Relative displacement is produced between imaging device；
Open position sensor 18, with the relative shift of real-time detection sample stage 14；
Using image processing apparatus 19, control light source 16 sends pulsed light, and controls imaging device at interval of a sampling
Interval time obtains a width low-resolution image of sample 13, and position sensing when recording acquisition every width low-resolution image
The relative shift of the sample stage 14 that device 18 is detected, all low point for then being obtained according to the relative shift of sample stage 14
Resolution imaging importing, synthesizes a panel height and differentiates using existing Image Restoration Algorithm (such as Fourier transform, deconvolution etc.)
The image of rate.
The amplitude of wherein described sample stage 14 is chosen to the maximum relative displacement between several low-resolution images
Size of the amount more than or equal to a pixel of the imageing sensor in imaging device.Between the sampling interval duration is selected as making
Relative shift between the adjacent low-resolution image of a sampling interval duration is less than one pixel of imageing sensor
2 times of size, the preferably equal to 1/10-1/3 of the size of one pixel of imageing sensor.
Microscope equipment that the present embodiment is provided and its imaging method and can obtain the height of big visual field without the need for the lens of high magnification numbe
Resolution image.Further, since light source can adopt parallel light source, therefore pattern distortion will not be produced.
Embodiment 2
The present embodiment provides a kind of microscope equipment for being adapted to detect for fluid sample, and its structure is as shown in fig. 6, be microscope equipment
Side view, the microscope equipment includes：
Light source 26, the light for being sent is directional light；
Sample stage 24, for carrying sample 23；
Imaging device, including lens 22 and imageing sensor 21, are fixedly installed on shaking platform 210, for obtaining
The image of the sample 23 under the irradiation of light source 26；
Vibrating device, including many springs 25 and vibrating motor 27, wherein vibrating motor 27 are used to make shaking platform 210 shake
It is dynamic, so as to generation is relative between the sample 23 that carries on sample stage 24 and hard-wired imaging device on shaking platform 210
Displacement, many springs 25 can be respectively fixed to each side of shaking platform 210, for making shaking platform 210 only in image
The plane internal vibration that sensor 21 is located；
Position sensor 28, the relative shift of the shaking platform 210 when vibrating for real-time detection；
Image processing apparatus 29, for controlling light source 26 pulsed light is sent, and controls imaging device acquisition sample 23
Several low-resolution images, and record the shaking platform that position sensor 28 is detected when obtaining every width low-resolution image
210 relative shift, is then superimposed all low-resolution images for obtaining according to the relative shift of shaking platform 210,
Synthesize a high-resolution image.Pulsed light obtains the low resolution of a width by can making imaging device at set intervals
Rate image, can easily obtain the output of several low-resolution images.
In the microscope equipment that the present embodiment is provided, light source 26, sample stage 24 and imaging device and the prior art for being adopted
In microscope equipment structure it is identical, need obtain low-resolution image when, can be as microscope equipment of the prior art
Directly the sample on sample stage 24 is imaged using imaging device.When needing to obtain high-definition picture, the present embodiment is provided
The imaging method of microscope equipment include：
The vibration frequency of vibrating motor 27 is set, shaking platform 210 is vibrated, so as to the sample carried on sample stage 24
23 and shaking platform 210 on produce relative displacement between hard-wired imaging device；
Open position sensor 28, with the relative shift of real-time detection shaking platform 210；
Using image processing apparatus 29, control light source 26 sends pulsed light, and controls imaging device at interval of a sampling
Interval time obtains a width low-resolution image of sample 23, and position sensing when recording acquisition every width low-resolution image
The relative shift of the shaking platform 210 that device 28 is detected, the institute that then will be obtained according to the relative shift of shaking platform 210
There is low-resolution image to be superimposed, such as using Fourier transform, the Image Restoration Algorithm such as deconvolution synthesizes a panel height resolution ratio
Image.
The amplitude of wherein described shaking platform 210 is chosen to the maximum between several low-resolution images with respect to position
Size of the shifting amount more than or equal to a pixel of the imageing sensor in imaging device.The sampling interval duration is selected as making
Relative shift between the adjacent low-resolution image of one sampling interval duration in interval is less than one picture of imageing sensor
2 times of the size of element, the preferably equal to 1/10-1/3 of the size of one pixel of imageing sensor.
Microscope equipment that the present embodiment is provided and its imaging method and can obtain the height of big visual field without the need for the lens of high magnification numbe
Resolution image.Further, since light source can adopt parallel light source, therefore pattern distortion will not be produced.
In the microscope equipment and its imaging method of the present embodiment offer, fixed sample stage, and imaging device is vibrated, therefore
It is specifically adapted for the sample that fluid sample etc. is not amenable to vibrate.
Embodiment 3
The present embodiment provides a kind of projection without lens microscope equipment, its structure as shown in fig. 7, comprises：
Light source 46, the light for being sent is directional light；
Sample cell 44, for carrying fluid sample 43；
Imageing sensor 41, is fixedly installed on shaking platform 410, for obtaining the sample 43 in the case where light source 46 irradiates
Image；
Vibrating device, including many springs 45 and vibrating motor 47, wherein vibrating motor 47 are used to make shaking platform 410 shake
It is dynamic, so as to produce between hard-wired imageing sensor 41 on the sample 43 and shaking platform 410 of carrying in sample cell 44
Relative displacement, many springs 45 are respectively fixed to each side of shaking platform 410, for making shaking platform 410 only in sample
The plane internal vibration that pond 44 is located；
Position sensor 48, the relative shift of the shaking platform 410 when vibrating for real-time detection；
Transparent lubriation material 42, between imageing sensor 41 and sample cell 44, for reduction rubbing between the two
Wipe the optical transfer characteristic of power and strengthening system；
The (not shown) of image processing apparatus 49, for controlling light source 46 pulsed light is sent, and controls imageing sensor
41 obtain several low-resolution images of sample 43, and position sensor 48 is detected when recording acquisition every width low-resolution image
The relative shift of the shaking platform 410 for arriving, all low resolution that then will be obtained according to the relative shift of shaking platform 410
Rate imaging importing, synthesizes a high-resolution image.
The present embodiment provide microscope equipment in, light source 46, sample cell 44 and the imageing sensor 41 for being adopted with it is existing
Structure of the projection in technology without lens microscope equipment is identical, when needing to obtain low-resolution image, can be as prior art
In projection equally directly the sample in sample cell 44 is imaged using imageing sensor 41 without lens microscope equipment.Needing
When obtaining high-definition picture, the imaging method of the microscope equipment that the present embodiment is provided includes：
The vibration frequency of vibrating motor 47 is set, shaking platform 410 is vibrated, so as to the sample carried in sample cell 44
43 and shaking platform 410 on produce relative displacement between hard-wired imageing sensor 41, at this moment transparent lubriation material 42
The frictional force between imageing sensor 41 and sample cell 44 can be reduced, and does not interfere with the imaging of imageing sensor 41；
Open position sensor 48, with the relative shift of real-time detection shaking platform 410；
Using image processing apparatus 49, control light source 46 sends pulsed light, and controls imaging device at interval of a sampling
Interval time obtains a width low-resolution image of sample 43, and position sensing when recording acquisition every width low-resolution image
The relative shift of the shaking platform 410 that device 48 is detected, the institute that then will be obtained according to the relative shift of shaking platform 410
There is low-resolution image to be superimposed, such as using Fourier transform, the Image Restoration Algorithm such as deconvolution synthesizes a panel height resolution ratio
Image.
The amplitude of wherein described shaking platform 410 is chosen to the maximum between several low-resolution images with respect to position
Size of the shifting amount more than or equal to a pixel of the imageing sensor in imaging device.The sampling interval duration is selected as making
Relative shift between the adjacent low-resolution image of one sampling interval duration in interval is less than one picture of imageing sensor
2 times of the size of element, the preferably equal to 1/10-1/3 of the size of one pixel of imageing sensor.
Microscope equipment that the present embodiment is provided and its imaging method and can obtain the height of big visual field without the need for the lens of high magnification numbe
Resolution image.Further, since light source can adopt parallel light source, therefore pattern distortion will not be produced.
In the microscope equipment and its imaging method of the present embodiment offer, fixed sample cell, and imageing sensor is vibrated, because
This is specifically adapted for the sample that fluid sample etc. is not amenable to vibrate.
Described " directional light " is not necessarily referring to the light of perfect parallelism in the present invention, also including less parallel light, art technology
Personnel are it is understood that perfect parallelism is just difficult to what is obtained, even the sunshine also not light of perfect parallelism, as
Sunshine is considered that directional light is the same, and be usually taken makes light source area larger in actual applications, or the distance of light source
Obtaining less parallel light, this means is also applied for the present invention to mode farther out.Described " directional light " refers to and sends out in the present invention
Scattered angle is the light beam of milliradian magnitude.
In above-described embodiment, the direction of vibration of sample stage or imaging device is limited by spring so as to only at its place
Plane internal vibration, in other embodiments of the invention, it would however also be possible to employ other stopping means limit sample stage or imaging dress
The direction of vibration put so as to only in a plane internal vibration.
In above-described embodiment, with transmission-type microscope equipment, (light that i.e. imaging device is detected crosses sample as lit transmissive
Light) as a example by describe the present invention.In other embodiments of the invention, it would however also be possible to employ (i.e. imaging is filled reflective microscope equipment
Put the reflected light that detected light is sample), can equally realize the present invention.
In above-described embodiment, pulse-like light source is employed, when being separated with certain sampling interval so as to obtain a series of
Between low-resolution image.In other embodiments of the invention, it would however also be possible to employ the light source for sending constant light of non-pulse formula,
And a series of acquisition of low-resolution images is realized by the shutter for arranging time switch before imageing sensor.
In the present invention, can adopt commonly used in the art the step of synthesize high-definition picture using several low-resolution images
The various algorithms for superimposed image, those skilled in the art can select according to actual needs and flexibly.Institute in the present invention
Using utilization several low-resolution images synthesize the algorithm of high-definition picture with of the prior art by " splicing "
Image and the method that obtains high-definition picture has difference substantially, concrete difference is as shown in Figure 8.Wherein Fig. 8 a are to pass through
" splicing " image and the method that obtains high-definition picture.Wherein 51 represent the high-definition picture seen in visual field (FOV), i.e.,
Image formed by imaging device, the position corresponding with formed image that 52 expression position sensors are sensed.Can be with from figure
It is clear that, the method for " splicing " image is only according to position by the corresponding high-definition picture in each position of alphabetical " A "
Put position that sensor senses and be stitched together.
And the utilization employed in the present invention several low-resolution images synthesize the signal of the algorithm of high-definition picture
Figure is as shown in Figure 8 b.In each width low-resolution image, all sites of alphabetical " A " are imaged (such as reference
Shown in 54), the position corresponding with formed image (as shown in reference 55) for then being sensed according to position sensor will
Several low-resolution image superpositions, finally synthesize a high-resolution image 56.It can thus be seen that the dress that the present invention is provided
Put and there is bigger visual field on the premise of the resolution ratio of equal size is obtained with method.
Fig. 9 shows the simulation result of microscope equipment of the invention and its imaging method.Using 9 width low resolution figures
As having synthesized a panel height image in different resolution.Relative shift between every width low-resolution image is the 1/3 of a Pixel Dimensions.
The resolution ratio of low-resolution image is about 1 micron, and the resolution ratio of high-definition picture is about 0.3 micron.
Figure 10 shows the another simulation result of microscope equipment of the invention and its imaging method.Using low point of 25 width
Resolution image has synthesized a panel height image in different resolution.Relative shift between every width low-resolution image is a Pixel Dimensions
1/3.The resolution ratio of low-resolution image is about 2 microns, and the resolution ratio of high-definition picture is about 0.4 micron.
In above-described embodiment, using position sensor come the relative shift between real-time detection sample and imaging device.
In other embodiments of the invention, it would however also be possible to employ other position sensing apparatus or method for sensing carry out detection sample with imaging
Relative shift between device.Present invention also offers a kind of location measurement method that can be used for microscope equipment, the method profit
With the reference marker (RP) on testing sample come the relative shift between detection sample and imaging device.
Figure 11 shows a kind of reference marker using on testing sample of present invention offer come detection sample and imaging dress
The location measurement method of the relative shift between putting.Wherein the reference marker is the line of L-shaped, and its live width depends on microscope equipment
Optical system, it is selected as making the live width of the reference marker of L-shaped projection to be on the image sensor equal to about picture
The size of element.As shown in figure 11, the reference marker on the image sensor L-shaped projection live width be W, equal to pixel 61,62,
63 and 64 width.
The vertical edge of the wherein L-shaped projection covers two of the 1st, 2 row in the first row in 3 × 3 picture element matrixs
Pixel 61 and 63, so as to form gray value g1 and g2 respectively in pixel 61 and 63, can be calculated by gray value g1 and g2
The left margin of reference marker is apart from the horizontal direction of the left margin of 3 × 3 picture element matrix apart from x.
The horizontal sides of L-shaped projection cover in the 3rd row in 3 × 3 picture element matrixs the 2nd, two pixels of 3 rows
62 and 64, so as to form gray value g3 and g4 respectively in pixel 62 and 64., reference can be calculated by gray value g3 and g4
The lower edge of mark is apart from the vertical direction of the lower boundary of 3 × 3 picture element matrix apart from y.
Wherein：
By measuring x values and y values, you can measure the relative shift between sample and imaging device.
Figure 12 shows another for providing of the invention using the reference marker on testing sample come detection sample and imaging
The location measurement method of the relative shift between device.Wherein the reference marker is square, and its length of side depends on micro- dress
The optical system put, it is selected as making the length of side of reference marker projection to be on the image sensor equal to about pixel
Size.As shown in figure 12, the length of side of reference marker projection on the image sensor is W, equal to pixel 65,66,67 and 68
Width.
Wherein the dimetric projection covers a part for 2 × 2 picture element matrixs, so as in pixel 65,66,67 and 68
Gray value g1, g2, g3, g4 are formed respectively, and the left margin distance of reference marker can be calculated by the gray value g1, g2, g3, g4
The horizontal direction of the left margin of 2 × 2 picture element matrix apart from x, and the lower edge of reference marker is apart from 2 × 2 pixel square
Battle array lower boundary vertical direction apart from y.
Wherein：
By real-time monitoring x value and y values, you can measure the relative shift between sample and imaging device.
Other embodiments of the invention, the shape of reference marker is not limited to above-mentioned L-shaped and square, it would however also be possible to employ
Other shapes.Furthermore it is also possible to measured using more individual pixel, for example with 4 × 4 picture element matrix.People in the art
Member can neatly select the shape of reference marker, and it is public that the calculating of relative shift is obtained by simple geometric transformation
Formula.
The microscope equipment and its imaging method that the present invention is provided is particularly suitable for detected size with the pixel of imageing sensor
The suitable testing sample of size, for example, the 100 of 1/10 to the pixel of pixel times, more preferably the 1 to 80 of pixel times.Especially fit
Determinand is, for example, the biological specimens such as red blood cell, leucocyte.
The invention also discloses a kind of imaging method of testing sample, the method includes：Obtain to be measured using imaging device
Sample and multiple image with first resolution；The testing sample is set to form relative displacement, institute with the imaging device
Bit shifts are in a plane；Detect the relative shift between the testing sample and the imaging device；Receive several
The image with first resolution and detected displacement, and there is first point by described according to detected displacement
The multiple image superposition of resolution, synthesizes the image with the second resolution higher than first resolution.
It should be understood that, although this specification is described according to each embodiment, but not each embodiment only includes one
Individual independent technical scheme, this narrating mode of specification is only that for clarity those skilled in the art will should say
Bright book as an entirety, the technical scheme in each embodiment can also Jing it is appropriately combined, forming those skilled in the art can be with
The other embodiment of understanding.
Schematically specific embodiment of the invention is the foregoing is only, the scope of the present invention is not limited to.It is any
Those skilled in the art, the equivalent variations made on the premise of the design without departing from the present invention and principle, modification and combination,
The scope of protection of the invention all should be belonged to.
Reference numerals list
Light source 6,06,16,26,46；Sample stage 4,14,24；Sample 13,23,43；Lens 12,22；Imageing sensor 01,
1、11、21、41；Spring 15,25,45；Vibrating motor 17,27,47；Position sensor 18,28；Image processing apparatus 19,29,
49；Shaking platform 210,410；Sample cell 04,44；Lubriation material 42；Camera lens 2；Image processing apparatus 5；Motor 7；Light-emitting component
06；Processor 05；Pixel 61,62,63,64,65,66,67,68.
Claims (14)
1. a kind of microscope equipment, including：
Gearshift, for making to produce relative displacement between testing sample and imaging device, institute's bit shifts are in a plane；
Imaging device, for obtaining multiple image of the testing sample in the various location relative to the imaging device,
Wherein described multiple image has first resolution;
Position sensing apparatus, for detecting the relative displacement between the testing sample and the imaging device；
Image processing apparatus, detect for receiving the multiple image with first resolution and the position sensing apparatus
The displacement, and described several are had by the imaging importing of first resolution according to the displacement for detecting, synthesis is provided
There is the image of the second resolution higher than first resolution.
2. microscope equipment according to claim 1, wherein the gearshift is used to be subjected to displacement the testing sample.
3. microscope equipment according to claim 1, wherein the gearshift is used to be subjected to displacement the imaging device.
4. microscope equipment according to claim 1, also includes the light for producing the directional light of the irradiation testing sample
Source.
5. microscope equipment according to claim 1, also includes the pulsed for sending the light for irradiating the testing sample
Light source.
6. microscope equipment according to claim 1, wherein the plane is horizontal plane.
7. microscope equipment according to claim 6, wherein the gearshift includes stopping means.
8. microscope equipment according to claim 1, wherein at interval of a sampling interval duration, the imaging device obtains
The image of testing sample described in width.
9. microscope equipment according to claim 8, wherein the sampling interval duration is selected as making described in one, interval
Relative shift between the adjacent image with first resolution of sampling interval duration is equal to the imaging device
A pixel size 1/10-1/3.
10. microscope equipment according to claim 1, wherein the displacement amplitude of the gearshift causes described several to have
Size of the maximum displacement between the image of first resolution more than a pixel of the imaging device.
11. microscope equipments according to claim 1, wherein the gearshift is vibrating device.
12. microscope equipments according to claim 1, wherein the microscope equipment is transmission-type or reflecting microscope, or
Person's projection is without lens microscope equipment.
13. microscope equipments according to claim 1, wherein the position sensing apparatus include being arranged on testing sample
Reference marker, reference marker imaging on the imaging device covers multiple pixels of the imaging device, according to
Relativeness between the gray value of each pixel in the plurality of pixel determines reference marker relative to the imaging device
Position.
A kind of 14. imaging methods, including：
Obtain testing sample using imaging device and multiple image with first resolution；
The testing sample is set to form relative displacement with the imaging device, institute's bit shifts are in a plane；
Detect the relative shift between the testing sample and the imaging device；
Receive several images with first resolution and detected displacement, and according to detected displacement by institute
The superposition of the multiple image with first resolution is stated, synthesizes the image with the second resolution higher than first resolution.
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| CN111819836B (en) * | 2018-03-14 | 2021-06-18 | 株式会社富士 | Imaging unit and component mounting machine |
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