CN106908942A - The parallel microscopic imaging apparatus of high-resolution based on microlens array - Google Patents

The parallel microscopic imaging apparatus of high-resolution based on microlens array Download PDF

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Publication number
CN106908942A
CN106908942A CN201710266273.4A CN201710266273A CN106908942A CN 106908942 A CN106908942 A CN 106908942A CN 201710266273 A CN201710266273 A CN 201710266273A CN 106908942 A CN106908942 A CN 106908942A
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array
hot spot
parallel
lens
microlens array
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唐玉国
肖昀
张运海
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • G02B21/0024Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/36Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365Control or image processing arrangements for digital or video microscopes

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  • Physics & Mathematics (AREA)
  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

The parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided, are designed to lighting module and generate the array light field comprising a large amount of hot spots with using microlens array, it is achieved thereby that the parallel illumination of sample, improves scanning imagery speed;Meanwhile, the parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided using planar array detector receiving array hot spot, and carry out pixel reallocation treatment to each hot spot in array hot spot, realize the parallel micro-imaging of high-resolution, high s/n ratio.

Description

The parallel microscopic imaging apparatus of high-resolution based on microlens array
Technical field
The present invention relates to microscopic detection instrument design and manufacture field, more particularly, to a kind of height based on microlens array Differentiate parallel microscopic imaging apparatus.
Background technology
Spot scan Laser Scanning Confocal Microscope has been widely used for the research fields such as biomedicine, is essential scientific research work Tool.Current shopping center's scanning confocal microscope mostly carries out simple scan imaging using galvanometer, but its simple scan into As rate limitation spot scan Laser Scanning Confocal Microscope further popularization and application in the fields such as living cells.
In recent years, many technologies are suggested, the image taking speed for improving spot scan Laser Scanning Confocal Microscope.But, this A little technologies usually improve image taking speed to sacrifice other performances such as resolution ratio, still can not be under the premise of resolution ratio is ensured well Solve slow this problem of image taking speed of spot scan Laser Scanning Confocal Microscope.
The content of the invention
The purpose of the present invention is:
A kind of high resolution and the fast parallel microscopic imaging apparatus of the high-resolution based on microlens array of presentation speed are provided.
To achieve the above object, the present invention uses following technical proposals:
A kind of parallel microscopic imaging apparatus of high-resolution based on microlens array, including lighting module, scan module, detection mould Block, control module and image reconstruction module, wherein:
The lighting module includes laser, the first lens, the second lens, microlens array and the 3rd lens, described micro- Lens array is made up of some lenticules, and the laser beam of the laser emitting is through first lens and the second collimated Formed after expanding and expand light beam, described expanding after light beam is again incident on the microlens array forms multiple array focal beam spots, The light beam that the multiple array focal beam spot sends forms multi beam collimated light beam after the 3rd collimated again;
The scan module includes dichroic mirror, object lens and nanometer displacement platform, and the nanometer displacement platform can be in XYZ three-dimensionals It is mobile, detected sample is carried on the nanometer displacement platform, the multi beam collimated light beam enters to inject after the dichroic mirror The object lens, and interfered at the front focal plane of the object lens, the interference array light field with hot spot is formed, it is described with light The interference array light field of spot is illuminated parallel to the sample so that the sample produces parallel optical signal;
The detecting module includes band-pass filter, finder lens and planar array detector, and the parallel optical signal is through institute The band-pass filter, finder lens and planar array detector are sequentially entered again after stating object lens and the dichroic mirror, and described The photosurface of planar array detector forms array hot spot, and the planar array detector detects the array hot spot, and to the array light Spot carries out space filtering and the pixel to the array hot spot and carries out reallocation treatment, to form less array hot spot, and will The less array hot spot is converted into electric signal;
The control module is electrically connected at the planar array detector and the nanometer displacement platform, and the control module is used for Gather the electric signal;
Described image is rebuild module and is electrically connected at the control module, and described image rebuilds module according to the control mould Block gathers the detector and realizes the high-resolution based on interference array light field simultaneously with the electric signal of the nanometer displacement platform Row scanning imagery image reconstruction.
Wherein in some embodiments, a branch of incident beam expanded in light beam is formed after the microlens array Array focal beam spot, the quantity of the array focal beam spot is identical with the quantity of lenticule in the microlens array.
Wherein in some embodiments, the planar array detector is the magazine one kind of CCD or CMOS.
The present invention is using the advantage of above-mentioned technical proposal:
The present invention provide the parallel microscopic imaging apparatus of the high-resolution based on microlens array, lighting module is designed with Using microlens array array light field of the generation comprising a large amount of hot spots, it is achieved thereby that the parallel illumination of sample, raising is scanned into As speed;Meanwhile, the parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided are connect using planar array detector Receive array hot spot, and pixel reallocation treatment carried out to each hot spot in array hot spot, realize high-resolution, high s/n ratio and Row micro-imaging.
The parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided, are formed using microlens array Array light field and pixel reallocation technology, can improve the image taking speed and resolution ratio of spot scan microtechnic, while can also protect Card image has signal to noise ratio higher, improves picture quality, is conducive to spot scan microtechnic in subcellular structure observation, thin Application in terms of the researchs such as born of the same parents' dynamic process observation, is beneficial to the development of biomedical sector.
Brief description of the drawings
Fig. 1 is the parallel microscopic imaging apparatus structural representation of the high-resolution based on microlens array provided in an embodiment of the present invention Figure.
Fig. 2 (a) is the structural representation of the illumination array hot spot at sample;
Fig. 2 (b) is the structural representation of the detection array hot spot on planar array detector;
Fig. 2 (c) is the structural representation in single array hot spot and 1AU dummy pinholes region;
Fig. 2 (d) is the structural representation of the corresponding pixel region of 1AU dummy pinholes;
Fig. 2 (e) is by the new hot spot and the structural representation of new dummy pinhole after pixel reallocation;
Fig. 2 (f) is the structural representation by the corresponding pixel region of new dummy pinhole after pixel reallocation;
Fig. 2 (g) is the structural representation by the new array hot spot after pixel reallocation;
Fig. 3 is the relative position of sample array hot spot along (dotted line) sample after (solid) before Y-direction movement and movement, light Spot spacing is d, moving step pitch dsStructural representation.
Wherein:Lighting module 110, scan module 120, detecting module 130, control module 140, image reconstruction module 150, Laser 111, two-dimensional grating 112, diaphragm 113, wave plate 114 and lens 115, dichroic mirror 121, object lens 122, nanometer displacement platform 123。
Specific embodiment
Fig. 1 is refer to, is a kind of parallel micro-imaging of high-resolution based on microlens array provided in an embodiment of the present invention Instrument 100, including lighting module 110, scan module 120, detecting module 130, control module 140 and image reconstruction module 150.Its In:
The lighting module 110 include laser 111, the first lens 112, the second lens 113, microlens array 114 and 3rd lens 115, the microlens array 114 is made up of some lenticules.
Specifically, the laser beam of the outgoing of the laser 111 is collimated through the lens 113 of first lens 112 and second Formed after expanding and expand light beam, described expanding after light beam is again incident on the microlens array 114 forms multiple arrays focusing light Spot, the light beam that the multiple array focal beam spot sends forms multi beam collimated light beam, institute after being collimated through the 3rd lens 115 again State multi beam collimated light beam re-incident and enter scan module 210.
It is appreciated that a branch of incident beam expanded in light beam forms array after the microlens array 114 gathering Burnt hot spot, the quantity of the array focal beam spot is identical with the quantity of lenticule in the microlens array 114.
The scan module 120 includes dichroic mirror 121, object lens 122 and nanometer displacement platform 123, the nanometer displacement platform 123 Detected sample can be carried on XYZ three-dimensional direction movings, the nanometer displacement platform 123.
Specifically, the multi beam collimated light beam enters to inject the object lens 122 after the dichroic mirror 121, and described Interfered at the front focal plane of object lens 122, form the interference array light field with hot spot, the interference array light with hot spot Field is illuminated parallel to the sample so that the sample produces parallel optical signal.
It is appreciated that due to nanometer displacement platform 123 can by XY directions mobile example, realize high-precision parallel scan into Picture, improves scanning imagery speed.
The detecting module 130 includes band-pass filter 131, finder lens 132 and planar array detector 133.
Specifically, the parallel optical signal sequentially enters the band logical again after the object lens 122 and the dichroic mirror 121 Colour filter 131, finder lens 132 and planar array detector 133, and in the photosurface formation array of the planar array detector 133 Hot spot, the planar array detector 133 detects the array hot spot, and carries out space filtering and to the battle array to the array hot spot The pixel of row hot spot carries out reallocation treatment, to form less array hot spot, and the less array hot spot is converted into Electric signal.
Preferably, planar array detector 133 is the magazine one kind of CCD or CMOS.It is appreciated that the face battle array that the application is used Detector 133 is CCD or CMOS cameras, with many pixels, can excite fluorescence simultaneously with a large amount of hot spots in receiving array light field The fluorescence signal that sample sends, and it is translated into electric signal.
The control module 140 is electrically connected at the planar array detector 133, and further, the control module 140 is also The nanometer displacement platform 123 is electrically connected at, the control module 140 is used to control the movement of the nanometer displacement platform 140.
It is appreciated that the nanometer displacement platform 123 scanning during the control module 140 can be realized to scan module is controlled System, while the electric signal of planar array detector 133 can be gathered.
Described image is rebuild module 150 and is electrically connected at the control module 140, and described image rebuilds module 150 can root The planar array detector 133 is gathered according to the control module 140 to realize being based on the electric signal of the nanometer displacement platform 123 The high-resolution parallel scan image of interference array light field is rebuild.
The course of work of image reconstruction module described further below 150:
First, sample is illuminated parallel using the array light field comprising array hot spot, shown in such as Fig. 2 (a), by array The sample of hot spot illumination will form (the M aggregation of array hot spot after being imaged through object lens and finder lens in planar array detector photosurface Hot spot) image, shown in such as Fig. 2 (b), realize that the detection of parallel optical signal is collected.
Second, in order to realize high-resolution imaging, it is necessary to the hot spot to detecting is carried out at space filtering and pixel reallocation Reason, with each array spot center as origin, 1 Airy disc size of selection is (it is appreciated that the big I of Ai Li spots is according to actual need Ask adjustment) corresponding pixel region (N number of pixel) is used as the dummy pinhole of each array hot spot, and such as Fig. 2 (c) and (d) are shown;Will The light intensity I that each pixel is detected in dummy pinholeiCarry out si/ 2 (wherein, siIt is the distance of each pixel to spot center) move Position, forms less new hot spot, shown in such as Fig. 2 (e);
Further according to the size of new hot spot, (region of dummy pinhole is with new hot spot to reselect the dummy pinhole of certain size As a example by size, less dashed rectangle and Fig. 2 (f) are shown in such as Fig. 2 (e), Q pixel in correspondence planar array detector), by the void Intend the light intensity I that each pixel is detected in pin holekSuperposition, can obtain any corresponding high-resolution micro-image pixel in sample ValueThis process is pixel reprocessing process.
3rd, each the array hot spot on planar array detector is carried out into pixel reallocation treatment, forms less array Hot spot (shown in such as Fig. 2 (g), black dotted lines are former array hot spot, and solid hot spot is small new array hot spot), obtains many in sample The corresponding high-resolution micro-image pixel value of individual point, after all hot spots treatment have been calculated, will obtain a width pixel value and is spaced apart High-resolution micro-image.
4th, when being scanned to sample, as shown in figure 3, array hot spot is moved (after dotted line is for movement on sample Array hot spot), array facula position is constant on array detector, in the array light field of irradiating sample hot spot spacing be d, if along Y Direction moving displacement platform, mobile spacing is ds, often moving can once obtain the high-resolution micrograph that a width pixel value is spaced apart Picture, mobile d/ds- 1 time, d/d can be obtainedsThe high-resolution micro-image that width pixel value is spaced apart, if mobile in X direction d/ds- 1 displacement platform, it is also possible to obtain d/dsPanel height differentiates micro-image, by this d/dsThe high-resolution that width pixel value is spaced apart Micro-image is superimposed, and can obtain the complete high-resolution micro-image of a width.
The parallel microscopic imaging apparatus 100 of the high-resolution based on microlens array that the present invention is provided, its course of work is as follows:
The laser beam of the outgoing of the laser 111 shape after the collimator and extender of 112 and second lens of the first lens 113 Into light beam is expanded, described expanding after light beam is again incident on the microlens array 114 forms multiple array focal beam spots, described many The light beam that individual array focal beam spot sends forms multi beam collimated light beam after being collimated through the 3rd lens 115 again, the multi beam is put down Row light beam enters to inject the object lens 122 after the dichroic mirror 121, and is interfered at the front focal plane of the object lens 122, The interference array light field with hot spot is formed, the interference array light field with hot spot is illuminated parallel to the sample, So that the sample produces parallel optical signal, the parallel optical signal after the object lens 122 and the dichroic mirror 121 again successively Into the band-pass filter 131, finder lens 132 and planar array detector 133, and in the sense of the planar array detector 133 Smooth surface forms array hot spot, and the planar array detector 133 detects the array hot spot, and carries out space filter to the array hot spot Involve carries out reallocation treatment to the pixel of the array hot spot, to form less array hot spot, while and by the array Hot spot is converted into electric signal, and the control module 140 gathers the electric signal, and described image rebuilds module 150 according to the control Molding block 140 gathers the electric signal and realizes that the high-resolution parallel scan image based on interference array light field is rebuild.
The parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided, set to lighting module 110 Count with using array light field of the generation comprising a large amount of hot spots of microlens array 114, it is achieved thereby that the parallel illumination of sample, improves Scanning imagery speed;Meanwhile, the parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided are visited using face battle array The receiving array hot spot of device 133 is surveyed, and pixel reallocation treatment is carried out to each hot spot in array hot spot, realize high-resolution, height The parallel micro-imaging of signal to noise ratio.
The parallel microscopic imaging apparatus of the high-resolution based on microlens array that the present invention is provided, are formed using microlens array Array light field and pixel reallocation technology, can improve the image taking speed and resolution ratio of spot scan microtechnic, while can also protect Card image has signal to noise ratio higher, improves picture quality, is conducive to spot scan microtechnic in subcellular structure observation, thin Application in terms of the researchs such as born of the same parents' dynamic process observation, is beneficial to the development of biomedical sector.
The parallel microscopic imaging apparatus of certain high-resolution based on microlens array of the invention can also have various conversion and change Type, it is not limited to the concrete structure of above-mentioned implementation method.In a word, protection scope of the present invention should include those for this area Obvious conversion or replacement and remodeling for those of ordinary skill.

Claims (3)

1. parallel microscopic imaging apparatus of a kind of high-resolution based on microlens array, it is characterised in that including lighting module, scanning mould Block, detecting module, control module and image reconstruction module, wherein:
The lighting module includes laser, the first lens, the second lens, microlens array and the 3rd lens, the lenticule Array is made up of some lenticules, and the laser beam of the laser emitting is expanded through first lens and the second collimated Formed afterwards and expand light beam, described expanding after light beam is again incident on the microlens array forms multiple array focal beam spots, described The light beam that multiple array focal beam spots send forms multi beam collimated light beam after the 3rd collimated again;
The scan module includes dichroic mirror, object lens and nanometer displacement platform, and the nanometer displacement platform can be moved in XYZ three-dimensionals It is dynamic, detected sample is carried on the nanometer displacement platform, the multi beam collimated light beam enters to inject institute after the dichroic mirror Object lens are stated, and interfered at the focal plane of the object lens, form the interference array light field with hot spot, it is described with hot spot Interference array light field is illuminated parallel to the sample so that the sample produces parallel optical signal;
The detecting module includes band-pass filter, finder lens and planar array detector, and the parallel optical signal is through the thing The band-pass filter, finder lens and planar array detector are sequentially entered after mirror and the dichroic mirror again, and in face battle array The photosurface of detector forms array hot spot, and the planar array detector detects the array hot spot, and the array hot spot is entered Row space filtering and the pixel to the array hot spot carry out reallocation treatment, to form less array hot spot, and will be described Less array hot spot is converted into electric signal;
The control module is electrically connected at the planar array detector and the nanometer displacement platform, and the control module is used to gather The electric signal;
Described image is rebuild module and is electrically connected at the control module, and described image is rebuild module and adopted according to the control module Collect the detector and realize that the high-resolution based on interference array light field is swept parallel with the electric signal of the nanometer displacement platform Retouch image reconstruction.
2. parallel microscopic imaging apparatus of the high-resolution based on microlens array according to claim 1, it is characterised in that described Expand a branch of incident beam in light beam and array focal beam spot is formed after the microlens array, the array focal beam spot Quantity is identical with the quantity of lenticule in the microlens array.
3. parallel microscopic imaging apparatus of the high-resolution based on microlens array according to claim 1, it is characterised in that described Planar array detector is the magazine one kind of CCD or CMOS.
CN201710266273.4A 2017-04-21 2017-04-21 The parallel microscopic imaging apparatus of high-resolution based on microlens array Pending CN106908942A (en)

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Cited By (5)

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CN108152941A (en) * 2017-11-20 2018-06-12 北京航空航天大学 High speed optical super-resolution imaging system and method based on micro-nano lens array
CN110365916A (en) * 2019-08-13 2019-10-22 苏州瑞霏光电科技有限公司 The reflective micro image collection system of array
CN110836892A (en) * 2019-11-26 2020-02-25 清华大学 Diffraction super-resolution microscopic imaging method and system
CN112219104A (en) * 2018-05-28 2021-01-12 维也纳自然资源与生命科学大学 Method for determining a three-dimensional particle distribution in a medium
CN113251949A (en) * 2021-06-18 2021-08-13 三代光学科技(天津)有限公司 Method for generating single-point optical measurement path of micro-lens array surface shape

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108152941A (en) * 2017-11-20 2018-06-12 北京航空航天大学 High speed optical super-resolution imaging system and method based on micro-nano lens array
CN112219104A (en) * 2018-05-28 2021-01-12 维也纳自然资源与生命科学大学 Method for determining a three-dimensional particle distribution in a medium
CN112219104B (en) * 2018-05-28 2024-03-26 维也纳自然资源与生命科学大学 Method for determining the three-dimensional particle distribution in a medium
CN110365916A (en) * 2019-08-13 2019-10-22 苏州瑞霏光电科技有限公司 The reflective micro image collection system of array
CN110836892A (en) * 2019-11-26 2020-02-25 清华大学 Diffraction super-resolution microscopic imaging method and system
CN110836892B (en) * 2019-11-26 2021-06-11 清华大学 Diffraction super-resolution microscopic imaging method and system
CN113251949A (en) * 2021-06-18 2021-08-13 三代光学科技(天津)有限公司 Method for generating single-point optical measurement path of micro-lens array surface shape
CN113251949B (en) * 2021-06-18 2021-11-30 三代光学科技(天津)有限公司 Method for generating single-point optical measurement path of micro-lens array surface shape

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