CN113867116A - Sub-pixel displacement imaging method for lensless microscopy - Google Patents
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- G—PHYSICS
- G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
- G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
- G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
- G03H1/0005—Adaptation of holography to specific applications
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- G—PHYSICS
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- G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
- G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
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- G—PHYSICS
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- G03H2001/005—Adaptation of holography to specific applications in microscopy, e.g. digital holographic microscope [DHM]
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Abstract
The invention discloses a sub-pixel displacement imaging method for lensless microscopy, which adopts a laser and a transmission type blazed grating to form diffraction light to be used as a light source to irradiate a sample to be observed, an image sensor senses a hologram of the sample to be observed, and the hologram is displaced by changing the central wavelength of emergent light of the laser. The invention also discloses a lens-free microscopic sub-pixel displacement imaging system which comprises a laser, a transmission type blazed grating and an image sensor, wherein emergent light of the laser forms diffracted light after passing through the transmission type blazed grating and irradiates to a sample to be observed. The imaging method and the imaging system provided by the invention open up a new way for a pixel or sub-pixel displacement method, and can be applied to lensless microscopic imaging.
Description
Technical Field
The invention relates to the field of optical microscopic imaging methods, in particular to a sub-pixel displacement imaging method for lensless microscopy.
Background
In recent years, with the development of scientific technology, microscopic imaging technology has been remarkably advanced. In microscopic imaging, the spatial bandwidth product is a measure of the information carrying capacity of an imaging system, and how to break through the limit of the spatial bandwidth product gradually becomes the focus of scientific research. Compared with the traditional microscope with the lens, the microscope without the lens which is emerging at present removes the constraint of the lens, and the sample is directly placed on the photoelectric sensing array, so that the imaging system improves the space bandwidth product under the condition of reducing the cost.
However, the resolution of the lens-less imaging system is limited by the pixel size of the image sensor, and for an image sensor of a certain imaging area, the larger the pixel size, the less information can be received. In terms of the prior art, the method of sub-pixel displacement imaging can realize super-resolution imaging, and breaks through the limitation of pixel size on resolution.
Conventional methods of achieving pixel or sub-pixel displacement include: the methods of mechanically moving the position of the light source, building a light source array, changing the incident angle of the light beam by using a mirror surface and the like have extremely high mechanical alignment requirements, have larger volume, are difficult to build, and have poor control over the displacement angle. In order to overcome the problems of the conventional methods, some researchers have proposed new methods for realizing sub-pixel displacement.
Manon of the sons federal institute of technology, los, proposed a miniaturized sub-pixel displacement structure. By using the volume phase holographic grating, a plurality of holograms are recorded in the photopolymer with the same volume, and then different reference lights are used for illuminating to generate diffracted lights with different angles, so that the angle change of the illuminating light is realized, and the sub-pixel displacement is realized. However, the volume holographic grating with matched parameters needs to be prepared on site by an interferometer before use, each parameter needs to be precisely controlled when the volume holographic grating is prepared on site, and the diffraction efficiency is low and is only about 20-30%. Only one side of the photopolymer on the volume holographic grating is protected by a glass layer, and the other side of the photopolymer is likely to be scratched in the process of manufacturing the grating for many times, resulting in poor imaging uniformity.
Chinese patent CN110187582A proposes a method for realizing sub-pixel displacement by changing the exit position of e-light using liquid crystal material with electrically controlled birefringence effect. This method has the following disadvantages: if the background image caused by o light is separated by using a calculation method, extra calculation amount is increased, and certain influence is caused on image quality, and the imaging light intensity of the method is less than 50% of the total light intensity, so that the light efficiency utilization rate is low, and the method is not beneficial to building an ultra-low power consumption imaging system.
Disclosure of Invention
The invention aims to provide a sub-pixel displacement imaging method for lensless microscopy, which aims to solve the problem of low light efficiency utilization rate in the optical microscopy displacement imaging technology in the prior art.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a sub-pixel displacement imaging method for lensless microscopy is characterized in that an image sensor senses a sample to be observed irradiated by a light source, a hologram of the sample to be observed is acquired by the image sensor, a laser and a transmission type blazed grating are adopted, emergent light of the laser forms diffracted light by the transmission type blazed grating and is used as the light source to irradiate the sample to be observed, the central wavelength of the emergent light of the laser is changed to deflect the angle of the formed diffracted light, and the hologram obtained by the image sensor is further displaced.
Furthermore, the central wavelength of the emergent light of the laser is changed by changing the driving signal of the laser.
Furthermore, the central wavelength of the emergent light of the adopted laser is regularly changed along with the change of the driving signal.
Furthermore, the change drift amount of the central wavelength of the emergent light of the adopted laser is in a linear relation with the driving signal.
Furthermore, the transmission blazed grating enables emergent light of the laser to form first-order diffraction light as a light source.
Furthermore, the formed diffraction light is deflected regularly by controlling a driving signal of the laser, so that the holograms obtained by the image sensor generate regularly changed displacement to form a plurality of holograms with low resolution, and the plurality of holograms with low resolution are synthesized to obtain a super-resolution image.
Further, a pixel super-resolution algorithm is adopted to synthesize a plurality of low-resolution holograms.
The sub-pixel displacement imaging system for the lensless microscope comprises an image sensor, a laser and a transmission type blazed grating, wherein the back surface of the transmission type blazed grating is a plane, the front surface of the transmission type blazed grating is a grating groove surface, the sensing side surface of the image sensor and the grating groove surface of the transmission type blazed grating face a sample to be observed respectively, and the light emergent end of the laser faces the back surface of the transmission type blazed grating, so that emergent light of the laser forms diffracted light after passing through the transmission type blazed grating, the diffracted light irradiates the sample to be observed, and the image sensor acquires a hologram of the sample to be observed.
And further, the laser device also comprises a controllable electric signal driver which is electrically connected with the laser device so as to output a driving signal with adjustable size to the laser device.
The imaging method and the imaging system provided by the invention open up a new way for a pixel or sub-pixel displacement method, and can be applied to lensless microscopic imaging.
The method and the system have the advantages that the method and the system are easy to realize, and complex mechanical structures are not needed; compared with a volume holographic grating and an electric control birefringence liquid crystal material, the transmission type blazed grating has high diffraction efficiency which can generally reach more than 80%, and high light efficiency utilization rate, and is very favorable for constructing an ultra-low power consumption and compact imaging system. In addition, the transmission blazed grating and the laser matched with the blazed wavelength have commercial purchase channels, the performance parameters are stable, the uniformity of diffraction light is good, and the microscopic imaging quality is favorably improved.
Drawings
Fig. 1 is a schematic optical path diagram of a transmission blazed grating when the 1 st order diffracted light coincides with a blaze angle.
FIG. 2 is a schematic diagram of a sub-pixel shift structure.
Fig. 3 shows a top view of the placement between the transmissive blazed grating and the image sensor.
FIG. 4 is a schematic representation of the displacement of a hologram of a sample as the angle of the illumination beam is varied.
Detailed Description
The invention is further illustrated with reference to the following figures and examples.
The main devices related to the invention include a transmission type blazed grating, a Vertical Cavity Surface Emitting Laser (VCSEL) and an image sensor.
The vertical cavity surface emitting laser is characterized in that the central wavelength of emergent light can be changed by changing the driving current input into the vertical cavity surface emitting laser, and the driving current and the drift amount of the central wavelength are in a linear relation in a certain range.
The transmission type blazed grating is characterized in that a grating groove surface is not parallel to a grating back surface (the grating back surface is a plane), the central maximum of single groove surface diffraction light in the grating groove surface is separated from an interference zero-order main maximum among grooves, and the energy of the light is transferred from the interference zero-order main maximum and is concentrated on a certain level of spectrum, so that the blazed spectrum is realized.
The method of the invention forms diffraction light by emergent light of the vertical cavity surface emitting laser through the transmission type blazed grating as a light source to irradiate to a sample to be observed, and enables the image sensor to sense the sample to be observed, thereby enabling the image sensor to obtain the hologram. The beam of light from the VCSEL is directed at an angle from the back of the transmission blazed grating as shown in FIG. 1. In fig. 1, the emergent light 4 of the vertical cavity surface emitting laser is incident from the back of the transmission type blazed grating 7, is refracted after passing through the transmission type blazed grating 7, and the refracted light is perpendicular to each groove surface of the grating and is diffracted at each groove surface, so that diffracted light is formed through the groove surface of the transmission type blazed grating 7, wherein the invention mainly utilizes the formed first-order diffracted light 5 which is coincident with the normal line of the groove surface as a light source. In fig. 1, 1 and 2 are normal lines of the grating surface, 4 is incident light, 3 is a reverse extension line of the first-order diffracted light 5, and 6 is a grating blaze angle γ. The equation for grating diffraction is:
in the formula (1)The pitch of the grating is shown as,representing the angle between the light beam inside the grating and the normal to the grating surface,which represents the refractive index of the grating material,the order of the diffracted light is shown,is shown asThe angle between the order diffraction light and the normal of the grating surface,represents the center wavelength of the light emitted from the vertical cavity surface emitting laser.
The sub-pixel displacement imaging system composed of the vertical cavity surface emitting laser and the transmission type blazed grating is shown in fig. 2. In fig. 2, the light emitting end of the vertical cavity surface emitting laser 8 is aligned to the back surface of the transmissive blazed grating 7 at a certain angle, the grating groove surface of the transmissive blazed grating 7 faces the sample 9 to be observed, the sensing surface of the image sensor 10 also faces the sample 9 to be observed, and each groove surface of the grating groove surface is parallel to the corresponding surface of the sample 9 to be observed and the image sensor 10.
The arrow above the transmissive blazed grating 7 in fig. 2 indicates the incident light, and the two arrows below the transmissive blazed grating 7 indicate the direction of the first order diffracted light. When the center wavelength of incident light of the transmission blazed grating 7 (i.e., the exit light of the vertical cavity surface emitting laser 8) changes, the angle of the first-order diffracted light changes. Each groove surface of the transmission type blazed grating 7 is parallel to the image sensor 10, the groove direction (namely the extending direction of the back surface plane of the transmission type blazed grating 7) and the right-angle side of the image sensor 10 form 45 degrees, and the first-order diffraction light is approximately vertical to the image sensor 10.
Is composed ofIt can be known that the center wavelengthWhen changed, the exit angle of diffracted light of each stageCorresponding changes may also occur. When the control current of the VCSEL is properly adjusted, the central wavelength of incident light can be changed by a few tenths of nanometers, and the angle of first-order diffracted light can be slightly changed. As shown in fig. 3, fig. 3 is a plan view of the arrangement positions of the transmission blazed grating 7 and the image sensor 10, 11 in fig. 3 is a deflection direction of the first-order diffracted light, and 12 is a grating groove direction.
When the angle change of the first order diffracted light is fixed, the distance of image displacement depends on the distance between the sample and the camera, and the larger the distance, the larger the distance of image displacement, as shown in fig. 4. In fig. 4, 13 and 14 respectively indicate two first-order diffracted lights corresponding to two different incident light center wavelengths, 9 indicates a sample to be observed, and 10 indicates an image sensor. By properly adjusting the magnitude of the driving current, the hologram of the sample on the image sensor 10 can generate a series of small displacements which are approximately in an arithmetic progression. A series of low-resolution holograms are collected by the image sensor 10, and a plurality of images are synthesized by using a pixel super-resolution algorithm, so that a super-resolution image can be obtained.
The invention is further illustrated by setting the following parameters:
selecting a central wavelength ofWith a minimum drive current ofThe center wavelength of the emergent light corresponding to the lowest driving current is. As the driving current increases, the center wavelength of the emitted light also increases. The driving current varies within a range ofWhen the central wavelength of the emergent light is inAnd is approximately linear with the drive current.
The parameters of the selected transmission type blazed grating are as follows: angle of flareEach millimeter hasThe refractive index of the grating material is。
According to the structure shown in figure 2A pixel imaging system. The distance between the transmission type blazed grating and the image sensor is approximatelyThe grating plane and the sensor plane formingThe angle is that the groove surface of the grating is parallel to the image sensor, and the groove direction of the grating is formed by the right-angle edge of the image sensorAnd (4) an angle.
The refractive index of the back surface of the grating is expressed byWhereinIs the angle between the incident beam and the normal of the grating surface,is the angle between the light beam inside the grating and the normal of the grating surface,. From the formula of refractive index, the incident angle of incident light should be as high as when the 1 st order diffracted light shinesThe light emitted from the VCSEL should be aligned with the grating surfaceThe angle and the direction perpendicular to the direction of the grating lines is incident on the back of the blazed grating.
According to the grating diffraction equation shown in equation (1):
formula (II)InThe pitch of the grating is shown as,,representing the angle between the light beam inside the grating and the normal to the grating surface,the order of the diffracted light is shown,is shown asThe angle between the order diffraction light and the normal of the grating surface,representing the center wavelength of the incident light.
Adjusting the control current of the vertical cavity surface emitting laser toSo that the central wavelength of the output light beam isAt this time, the 1 st order diffracted light of the blazed grating is vertically irradiated on the image sensor. Above the image sensorAnd placing a sample to be observed, and recording a holographic image of the sample.
According to the formulaWhen the control current of the vertical cavity surface emitting laser rises toThe output center wavelength of the VCSEL is determined byIs changed intoThe angle of 1 st order diffraction light of blazed grating is aboutThe hologram of the sample to be observed on the image sensor is deflected approximately in the direction perpendicular to the grating linesDisplacement of (2). Because the change of the central wavelength of the output light of the vertical-cavity surface-emitting laser is very small, the change of the angle of the 1 st-order diffraction light is also very small, and the changes can be ignored when the hologram is reconstructed and a pixel super-resolution algorithm is used.
In thatIs arranged betweenStep size of change. Each change of drive currentThe angle of the 1 st order diffracted light will occurDeflection of, sampleCan be generatedCan acquire 11 holographic images which are approximately in arithmetic progression displacement. And a high-resolution image is restored from the series of low-resolution images by using a pixel super-resolution algorithm, so that the problem of undersampling caused by discrete pixels of the image sensor is solved.
The embodiments of the present invention are described only for the preferred embodiments of the present invention, and not for the limitation of the concept and scope of the present invention, and various modifications and improvements made to the technical solution of the present invention by those skilled in the art without departing from the design concept of the present invention shall fall into the protection scope of the present invention, and the technical content of the present invention which is claimed is fully set forth in the claims.
Claims (9)
1. A sub-pixel displacement imaging method for lens-free microscopy is characterized in that an image sensor senses a sample to be observed which is irradiated by a light source, and a hologram of the sample to be observed is acquired by the image sensor, wherein the method comprises the following steps: the method comprises the steps of adopting a laser and a transmission type blazed grating, enabling emergent light of the laser to form diffracted light serving as a light source to irradiate a sample to be observed by utilizing the transmission type blazed grating, and enabling the angle of the formed diffracted light to deflect by changing the central wavelength of the emergent light of the laser, so that a hologram obtained by an image sensor is displaced.
2. A sub-pixel displacement imaging method for lensless microscopy according to claim 1, wherein: the central wavelength of the emergent light of the laser is changed by changing the driving signal of the laser.
3. A sub-pixel displacement imaging method for lensless microscopy according to claim 2, wherein: the central wavelength of the emergent light of the adopted laser is regularly changed along with the change of the driving signal.
4. A sub-pixel displacement imaging method for lensless microscopy according to claim 3, wherein: the change drift amount of the central wavelength of the emergent light of the adopted laser is in a linear relation with the driving signal.
5. A sub-pixel displacement imaging method for lensless microscopy according to claim 1, wherein: the transmission type blazed grating enables emergent light of the laser to form first-order diffraction light to be used as a light source.
6. A sub-pixel displacement imaging method for lensless microscopy according to claim 1, wherein: the formed diffraction light is deflected regularly by controlling a driving signal of the laser, so that the holograms obtained by the image sensor generate regularly changed displacement to form a plurality of holograms with low resolution, and the plurality of holograms with low resolution are synthesized to obtain a super-resolution image.
7. The sub-pixel displacement imaging method for lensless microscopy according to claim 6, wherein: and synthesizing the plurality of low-resolution holograms by adopting a pixel super-resolution algorithm.
8. A sub-pixel displacement imaging system for lensless microscopy comprising an image sensor, characterized in that: the back surface of the transmission type blazed grating is a plane, the front surface of the transmission type blazed grating is a grating groove surface, the sensing side surface of the image sensor and the grating groove surface of the transmission type blazed grating face towards a sample to be observed respectively, the light emergent end of the laser faces towards the back surface of the transmission type blazed grating, therefore, emergent light of the laser forms diffracted light after passing through the transmission type blazed grating, the diffracted light irradiates the sample to be observed, and the image sensor acquires a hologram of the sample to be observed.
9. A sub-pixel displacement imaging system for lensless microscopy according to claim 8, wherein: the laser device also comprises a controllable electric signal driver which is electrically connected with the laser device so as to output a driving signal with adjustable size to the laser device.
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CN115995756A (en) * | 2023-03-23 | 2023-04-21 | 苏州长光华芯半导体激光创新研究院有限公司 | Wavelength locking system and wavelength locking method |
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