CN108802994B - Image recording system and method - Google Patents
Image recording system and method Download PDFInfo
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- CN108802994B CN108802994B CN201810581373.0A CN201810581373A CN108802994B CN 108802994 B CN108802994 B CN 108802994B CN 201810581373 A CN201810581373 A CN 201810581373A CN 108802994 B CN108802994 B CN 108802994B
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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
- G02B21/361—Optical details, e.g. image relay to the camera or image sensor
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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
- G02B21/362—Mechanical details, e.g. mountings for the camera or image sensor, housings
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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
- G02B21/368—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements details of associated display arrangements, e.g. mounting of LCD monitor
Abstract
The application relates to a microscopic image recording system and a microscopic image recording method. The recording system divides light emitted by a laser into two beams of light waves, wherein one beam of light waves carries information of an object, and the other beam of light waves is expanded and collimated and then passes through a microscope objective to form spherical light waves; two beams of light waves are interfered on the CCD after passing through the beam combining mirror to form a hologram, and the interference pattern is reproduced through a computer so as to obtain a three-dimensional microscopic image of the object. The computer can automatically control the rotation of the beam combining mirror, so that the core position of the spherical light wave is adjusted, the original image and the middle interference item in the obtained reproduced image are just separated, and the reproduced image with high quality is obtained.
Description
Technical Field
The application relates to the recording of microscopic images, and belongs to the field of optical imaging.
Background
The conventional microscope is mainly obtained through a lens group, so that the obtained microscopic image cannot be recorded and stored, and the details of the image after observation are quickly unclear. With the continuous development of semiconductor technology and laser technology, a digital microscope is appeared at present, which can obtain a phase image of an imaged object, or a three-dimensional image of the imaged object, and can permanently store the obtained three-dimensional image while obtaining an intensity image of the imaged object.
Digital microscopy, also known as digital holography, uses a CCD to collect a hologram of an imaged object, inputs the hologram into a computer, and uses an algorithm to simulate the actual hologram reconstruction process in the computer, thereby reconstructing three of the object in the computerAnd (5) dimension image. Compared with the image acquired by a common digital camera, the reconstructed image of the hologram has a phase image, namely a three-dimensional appearance image of an object, besides an intensity image, so that the reconstruction of the phase information of the object is the most critical in digital holography. In the existing digital holography, most of the off-axis holography is adopted to obtain a digital holographic image, in a recording system of the off-axis holography, laser emitted from a laser is divided into two beams of light by a beam splitter, one beam of light carries object information (called as crop light wave) after passing through an object (sample), and reaches a CCD target surface after passing through a beam combiner BS, and the other beam of light (called as reference light wave) is expanded and collimated and then passes through a reflector M2And the reflected light is reflected by a beam combiner and reaches a CCD target surface to interfere with object light waves to form an interference image (namely a hologram). In the prior art, the off-axis digital holography is mainly divided into off-axis Fresnel digital holography and off-axis lens-free Fourier transform digital holography, in the recording of the off-axis Fresnel digital holography, reference light adopts planar light waves, and when the digital holography is recorded by using the planar reference light waves, the structure of a recording system determines that the frequency of interference fringes in a partial region in an interference light field is low, the frequency of the partial region is high, so that the bandwidth of a CCD (charge coupled device) cannot be fully utilized, the recording distance is limited by the size of a CCD photosensitive surface, and the resolution of a reproduced image is difficult.
Disclosure of Invention
The application provides a microscopic imaging system, which adopts spherical reference light waves to record digital holograms, thereby improving the resolution of reproduced images.
The following technical scheme is adopted in the application: a microscopic image recording system comprises a laser 1, a first half-wave plate 2, a polarization beam splitter 3, a first reflector 4, a first beam expanding collimator 5, a transparent object 6, a beam combiner 7 (semi-transmission and semi-reflection), a CCD8, a second half-wave plate 9, a second beam expanding collimator 10, a second reflector 11, a microscopic objective 12 and a computer 13; a light beam emitted by the laser is divided into a light beam A and a light beam B after passing through the polarization beam splitter, the light beam A enters the first beam expanding collimating mirror after passing through the first reflector to form parallel light and irradiate a transparent object to form an object light wave, and the object light wave reaches a CCD target surface after penetrating through the beam combining mirror; the light beam B after passing through the polarization beam splitter forms a plane light wave through a second half-wave plate and a second beam expanding collimating lens 10, the plane light wave enters the microscope objective through a second reflecting lens to form a spherical reference light wave, the center of the spherical reference light wave is a reference point light source, and the spherical reference light wave reaches a CCD target surface after passing through the beam combining lens to form a holographic image with the object light wave; the hologram recorded by the CCD is transmitted to a computer and stored, the computer can automatically control the rotation of the beam combining mirror, so as to control the position of the reference point light source to obtain the optimal hologram, and the original image, the conjugate image and the intermediate interference item in the reconstructed image which can be obtained after the interference pattern is reconstructed are just separated. The method is characterized in that: still include angle governing system, angle governing system includes: the device comprises a supporting platform, a rotating shaft, a pull rope, a displacement detector and a driving motor; the beam combining mirror is fixed on the supporting platform and is driven to rotate through rotation of the rotating shaft, so that the position of the reference point light source on an object plane is adjusted, the rotation of the rotating shaft drives displacement of the pull rope, the moving distance of the pull rope is monitored through the displacement detector, the moving distance of the pull rope is obtained through the displacement detector, and then the rotating angle of the rotating shaft is obtained.
According to the method, the spherical reference light wave is adopted to record the hologram, the fringe space frequency of an interference field is relatively low, so that the sampling condition for recording the digital hologram is easy to meet, and especially when a recording light path is arranged according to a method of lens-free Fourier transform holography, due to the fact that interference fringes are approximately parallel and equal in distance, the limited bandwidth of a CCD can be fully utilized. The allowable minimum recording distance is not limited by the size of the CCD, and the hologram can be recorded in a small distance for a tiny object to obtain more information, so that the improvement of the resolution of a reproduced image is facilitated, and therefore, the recording of the lens-free Fourier transform digital hologram by using the spherical reference light wave is an effective way for realizing high-resolution imaging.
Drawings
FIG. 1: coordinate schematic diagram of the spherical reference light wave digital hologram record;
FIG. 2: an off-axis lensless Fourier transform digital holographic recording system;
FIG. 3: an angle adjustment automatic control system.
Detailed Description
The recording path of a spherical reference wave digital hologram and the coordinate system used for the analysis are shown in FIG. 1, where x0-y0The plane is an object plane, the x-y plane is a hologram plane, the z axis vertically passes through the centers of the two planes, and the position coordinate of the reference point source is (x)r,yr,zr) Wherein z isrRepresenting the distance of the point source to the CCD plane. In practical applications, z is often takenr>z0。
According to the Fresnel diffraction formula, under the condition of paraxial approximation, constant phase factors are ignored, and object light waves and reference light waves reaching a CCD plane are respectively
The term representing the original image in the hologram is UO*After finishing to obtain
Wherein
The phase of each point in the diffraction field and the spatial frequency of the fringes along the x and y directions are respectively
Considering z0、zrAre all positive, and zr≥z0Then hologramFringe maximum and minimum spatial frequencies of
According to the Nyquist's theorem of sampling
From equations (1-8) and (1-9), the constraints imposed on the reference light bias to satisfy the sampling theorem can be derived as follows:
halo light | U2Has a spatial frequency in the x and y directions of which the maximum values are respectively
In order to separate the reproduced images from each other, it is only necessary to appropriately set the position of the reference light so that the frequency spectrums of the zero-order diffraction term, the original image, and the conjugate image do not overlap with each other, that is, it is required that
Thereby obtaining another limit condition set for the position of the reference light source in the off-axis holography of the spherical reference light wave, namely the separation condition of the reproduced image is
The position of the reference point source can be determined by combining the formula (1-10), wherein zrIs obtained by enlarging the reproduced imageThe magnification requirement is determined by the form of the reproduced reference light wave.
In the method for recording the hologram by utilizing the spherical reference light wave, off-axis lensless Fourier transform holography and on-axis phase shift lensless Fourier transform holography are applied more frequently. The off-axis optical path has the advantages that the zero-order diffraction image and the positive-negative first-order diffraction image are separated from each other, interference items are filtered easily, and the single hologram can be used for reconstruction. Off-axis lensless fourier transform holography is discussed below.
Off-axis lensless fourier transform holography is one of the commonly used recording optical path structures in digital holography. An off-axis lensless Fourier transform digital holographic recording system is shown in FIG. 2, and comprises a laser 1, a first half-wave plate 2, a polarization beam splitter 3, a first reflector 4, a first beam expanding collimator 5, a transparent object 6, a beam combiner 7 (semi-transmission and semi-reflection), a CCD8, a second half-wave plate 9, a second beam expanding collimator 10, a second reflector 11, a microscope objective 12 and a computer 13; a light beam emitted by the laser is divided into a light beam A and a light beam B after passing through the polarization beam splitter, the light beam A enters the first beam expanding collimating mirror after passing through the first reflector to form parallel light and irradiate a transparent object to form an object light wave, and the object light wave reaches a CCD target surface after penetrating through the beam combining mirror; the light beam B after passing through the polarization beam splitter forms a plane light wave through a second half-wave plate and a second beam expanding collimating lens 10, the plane light wave enters the microscope objective through a second reflecting lens to form a spherical reference light wave, the center of the spherical reference light wave is a reference point light source, and the spherical reference light wave reaches the CCD target surface after passing through the beam combining lens and forms a holographic image with the object light wave. When the recording reference point source is located on the object plane, zr＝z0Substituting the equations (1-10) and (1-13) to obtain the bias requirement satisfying both the sampling condition and the reproduced image separating condition
Wherein the equal sign represents the critical separation and critical sampling, and the minimum recording distance can be obtained by solving
z0min＝max(4X/λΔx,4Y/λΔy) (1-15)
In the formula, max represents the larger value of the two.
From the above analysis, it can be seen that the separation degree of the reconstructed image of the lensless fourier transform hologram is related to the coordinates of the reference point light source, and does not relate to the angle between the object light wave and the reference light wave. However, through the analysis of the recording optical path, it can be known that the reference point light source and the object represented by the microscope objective are mirror-symmetric with respect to the half-transmitting mirror surface of the beam combiner, that is, the reference point light source and the object are located on the same plane through the imaging of the half-transmitting mirror surface, and in the process of the optical path arrangement, since the object light wave and the reference light wave are both arranged in parallel mesas, that is, the y coordinate of the reference point light source can be considered as zero, at this time, the separation degree of the reproduced image of the lensless fourier transform hologram is only related to the x coordinate of the reference point light source, and the x coordinate of the point light source can be changed by rotating the beam combiner, thereby affecting the separation degree of the reproduced. In order to obtain a sufficiently separated reproduction image with the maximum resolution, a method of precisely adjusting the coordinates of the reference point light source is described below; the method specifically comprises the following steps:
1) measuring a first distance d from the beam combiner to the CCD target surface1And recording;
2) collecting and storing first holograms of the object light wave and the reference light wave;
3) reconstructing the first hologram to obtain a reproduced image 1, and judging the separation degree of an original image and an intermediate interference item in the obtained reproduced image 1;
4) if the original image and the intermediate interference item in the reproduced image 1 are overlapped or partially overlapped, the computer sends an instruction to control the beam combining mirror to rotate by a specified angle (for example, 0.1 degree);
5) continuously collecting and storing second holograms of the object light wave and the reference light wave;
6) reconstructing the second hologram to obtain a reproduced image 2, and judging the separation degree of the original image and the intermediate interference item in the obtained reproduced image 2;
7) if the original image and the intermediate interference item in the reproduced image 2 are still partially overlapped, repeating the steps 4) -6) until the original image and the intermediate interference item in the reproduced image are completely separated;
8) continuously judging the reproduced image with the original image and the intermediate interference item completely separated, and if the distance between the original image and the intermediate interference item is too far, sending an instruction by the computer to control the half-reflecting and half-transmitting mirror to rotate by a specified angle (for example, 0.05 degrees) in the reverse direction;
9) continuously collecting and storing third holograms of the object light wave and the reference light wave;
10) reconstructing the third hologram to obtain a reproduced image 3, and judging the separation degree of the original image and the intermediate interference item in the obtained reproduced image 3;
11) if the distance between the original image and the intermediate interference item is still too far, repeating the above steps 8) -10) until the original image and the intermediate interference item in the reproduced image are just separated; the corresponding hologram at this time is d1Distance-corresponding optimal hologram H1(ii) a Subjecting the optimal hologram H1Storing in a database;
12) controlling the CCD to move forwards or backwards for a specified distance delta d, and then repeating the steps 1) -11) until the distance d is obtained1Optimum hologram H corresponding to + -Delta d2The optimal hologram H2Stored in a database.
13) The best hologram H to be obtained1、H2…HnTo a corresponding distance d1、d2…dnAnd establishing a distance-hologram corresponding table.
14) In the subsequent recording of the hologram, firstly measuring the distance d from the CCD to the beam combining mirror, inputting the distance d, and calling out the required optimal hologram H from the distance-hologram corresponding table in the computer to be used as a standard hologram; secondly, the computer sends out an instruction to control the rotation angle of the beam combining mirror and then shoots a hologram M, the hologram M is matched (namely identified) with the standard hologram, and if the holograms are not matched, the rotation angle is continued to shoot the hologram; if the holograms match, the hologram M taken at this time is saved as a hologram for reconstructing a three-dimensional image of the object. In the process of matching the holograms, because it is difficult to completely match the two holograms, an error threshold may be set, and when the matching error value is smaller than the threshold, the two holograms may be considered as matched, and when the matching error value is larger than the threshold, the two holograms may not be matched.
According to the method, the optimal hologram corresponding to the distance is collected, the distance-hologram corresponding table is established, and in the later adjustment of the digital microscope system, only the distance from the CCD to the beam combining mirror needs to be input, the computer system automatically controls the beam combining mirror to rotate and obtain the optimal hologram, and a more accurate three-dimensional reconstruction image can be obtained through the optimal hologram.
Furthermore, the application also provides an angle adjusting system for accurately controlling the rotation angle of the beam combining mirror through a computer. As shown in fig. 3, the angle adjusting system includes: support platform 14, rotation axis 15, stay cord 16, displacement detector 17, driving motor 18. The beam combining mirror is fixed on the supporting platform, the beam combining mirror is driven to rotate through rotation of the rotating shaft, so that the position of the reference point light source on an object plane is adjusted, the rotation of the rotating shaft drives displacement of the pull rope, the moving distance of the pull rope is monitored through the displacement detector, the moving distance of the pull rope is obtained through the displacement detector, and then the rotating angle of the rotating shaft is obtained. The computer 13 controls the driving motor to drive the rotating shaft to rotate, the stay cord is driven to displace in the rotating process of the rotating shaft, the displacement detector detects the linear displacement of the stay cord, when the linear displacement is equal to a preset value, the displacement detector sends a signal to the computer to control the rotating shaft to stop rotating, and the preset value corresponds to a specified angle at which the beam combiner needs to rotate. Through the angle adjusting system, the position of the reference point light source in the off-axis lens-free Fourier transform digital holographic record can be accurately adjusted, and the separation degree and resolution of the reproduced image are controlled, so that the reproduced image with high quality is obtained.
Claims (3)
1. A microscopic image recording method for microscopic image recording in a microscopic image recording system, the microscopic image recording system comprising: the device comprises a laser, a first half-wave plate, a polarization beam splitter, a first reflector, a first beam expanding collimator, a transparent object, a beam combining mirror, a CCD (charge coupled device), a second half-wave plate, a second beam expanding collimator, a second reflector, a microscope objective and a computer; a light beam emitted by the laser is divided into a light beam A and a light beam B after passing through the polarization beam splitter, the light beam A enters the first beam expanding collimating mirror after passing through the first reflector to form parallel light and irradiate a transparent object to form an object light wave, and the object light wave reaches a CCD target surface after penetrating through the beam combining mirror; the light beam B after passing through the polarization beam splitter forms a plane light wave through a second half-wave plate and a second beam expanding collimating lens, the plane light wave enters the microscope objective through a second reflecting lens to form a spherical reference light wave, the center of the spherical reference light wave is a reference point light source, and the spherical reference light wave reaches the CCD target surface after passing through the beam combining lens to form a hologram with the object light wave; the hologram recorded by the CCD is transmitted to a computer and stored, the computer automatically controls the rotation of the beam combining mirror, so that the relative position of the reference point light source and the plane of the transparent object is controlled to obtain the optimal hologram, and the original image, the conjugate image and the intermediate interference item in the reconstructed image which can be obtained by the optimal hologram are just separated; the microscopic image recording system further comprises an angle adjustment system, the angle adjustment system comprising: the device comprises a supporting platform, a rotating shaft, a pull rope, a displacement detector and a driving motor; the beam combining mirror is fixed on the supporting platform and is driven to rotate through the rotation of the rotating shaft, so that the relative position of the reference point light source on the object plane is adjusted; the rotation of the rotating shaft drives the stay cord to move, the moving distance of the stay cord is monitored by a displacement detector, the moving distance of the stay cord is obtained by the displacement detector, and then the rotating angle of the rotating shaft is obtained; the method is characterized in that: the microscopic image recording method comprises the following steps:
1) measuring a first distance d from the beam combiner to the CCD target surface1And recording;
2) collecting and storing first holograms of the object light wave and the reference light wave;
3) reconstructing the first hologram to obtain a first reproduced image, and judging the separation degree of an original image and an intermediate interference item in the obtained first reproduced image;
4) if the original image in the first reproduced image and the intermediate interference item are completely overlapped or partially overlapped, the computer sends an instruction to control the beam combining mirror to rotate by a specified angle;
5) continuously collecting and storing second holograms of the object light wave and the reference light wave;
6) reconstructing the second hologram to obtain a second reproduced image, and judging the separation degree of the original image and the intermediate interference item in the obtained second reproduced image;
7) if the original image and the intermediate interference item in the reproduced image II are still partially overlapped, repeating the steps 4) -6) until the original image and the intermediate interference item in the reproduced image are completely separated;
8) continuously judging the reproduced image with the original image and the intermediate interference item completely separated, and if the distance between the original image and the intermediate interference item is too far, sending an instruction by the computer to control the beam combiner to rotate in the opposite direction by half of a specified angle;
9) continuously collecting and storing third holograms of the object light wave and the reference light wave;
10) reconstructing the third hologram to obtain a third reproduced image, and judging the separation degree of the original image and the intermediate interference item in the third reproduced image;
11) if the distance between the original image and the intermediate interference item is still too far, repeating the above steps 8) -10) until the original image and the intermediate interference item in the reproduced image are just separated; the corresponding hologram at this time is d1Distance-corresponding optimal hologram H1(ii) a Subjecting the optimal hologram H1Storing in a database;
12) controlling the CCD to move forwards or backwards for a specified distance delta d, and then repeating the steps 1) -11) until the distance d is obtained1Optimum hologram H corresponding to + -Delta d2The optimal hologram H2Storing in a database;
13) the best hologram H to be obtained1、H2…HnTo a corresponding distance d1、d2…dnEstablishing a distance-hologram corresponding table;
14) in the subsequent recording of the hologram, firstly measuring the distance d from the CCD to the beam combining mirror, inputting the distance d, and calling out the required optimal hologram H from the distance-hologram corresponding table in the computer to be used as a standard hologram; secondly, the computer sends out an instruction to control the rotation angle of the beam combining mirror and then shoots a hologram M, the hologram M is matched with the standard hologram, and if the holograms are not matched, the rotation angle is continued to shoot the hologram; if the holograms match, the hologram M taken at this time is saved as a hologram for reconstructing a three-dimensional image of the object.
2. A microscopic image recording method according to claim 1, characterized in that: the computer controls the driving motor to drive the rotating shaft to rotate, the stay rope is driven to displace in the rotating process of the rotating shaft, the displacement detector detects the linear displacement of the stay rope, when the linear displacement is equal to a preset value, the displacement detector sends a signal to the computer to control the rotating shaft to stop rotating, and the preset value corresponds to a specified angle at which the beam combiner needs to rotate.
3. A microscopic image recording method according to claim 1, characterized in that: in the process of matching the holograms, an error threshold is set, and when the error value of the matching is smaller than the threshold, the matching is considered.
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| CN102749834A (en) * | 2012-07-24 | 2012-10-24 | 河北工程大学 | Optical microscopic imaging system and imaging method |
| CN104024835A (en) * | 2011-11-17 | 2014-09-03 | 东丽工程株式会社 | Automatic visual inspection device |
| US9541374B2 (en) * | 2007-12-31 | 2017-01-10 | Stc.Unm | Structural illumination and evanescent coupling for the extension of imaging interferometric microscopy |
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| US9541374B2 (en) * | 2007-12-31 | 2017-01-10 | Stc.Unm | Structural illumination and evanescent coupling for the extension of imaging interferometric microscopy |
| CN104024835A (en) * | 2011-11-17 | 2014-09-03 | 东丽工程株式会社 | Automatic visual inspection device |
| CN102749834A (en) * | 2012-07-24 | 2012-10-24 | 河北工程大学 | Optical microscopic imaging system and imaging method |
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