CN107703642B - single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device and using method - Google Patents
single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device and using method Download PDFInfo
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Abstract
the invention discloses a single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device and a using method thereof, wherein the device comprises a pulse laser (1), an attenuation sheet (2), a polarizer (3), a first lens (4), a diaphragm (5), a sample table (6), a reflector group (7), a reflector with holes (8) and a light beam collector (9) which are coaxially arranged in sequence along the advancing direction of a light beam; a second lens (10) and a liquid nitrogen cooling CCD image sensor (11) are sequentially arranged along the advancing direction of the reflected light beam of the reflector group (7); and a common CCD image sensor (12) is arranged in the forward direction of a reflected light beam of the reflector (8) with the hole, and the liquid nitrogen cooling CCD image sensor (11) and the common CCD image sensor (12) are both connected with a computer (13). By adopting the device, high-angle and central diffraction light spots can be obtained simultaneously, and the three-dimensional reconstruction precision is improved.
Description
Technical Field
The invention belongs to the field of laser coherent diffraction imaging, and particularly relates to a single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device and a using method thereof.
Background
Coherent diffraction imaging utilizes advanced light sources, such as synchrotron radiation light sources, higher harmonic light sources, X-ray lasers, free electron lasers and visible light lasers, in combination with a phase recovery algorithm to calculate the diffraction image back as a structural image of the sample. The method has the advantages that the imaging sample does not need to be calibrated, and the method can be quantitative and has higher resolution. The imaging method has realized two-dimensional and three-dimensional imaging of various nanomaterials and biological cells.
three-dimensional imaging plays an important role in understanding the relationship between materials and biological tissue structure and function. For an electron microscope, when a sample is thick, a single-layer image is obtained through slicing, and then a three-dimensional image is combined, and when the sample is small, the three-dimensional image is obtained through rotating the sample; for a confocal microscope, a three-dimensional image is obtained by focusing light beams at different depths of a sample and scanning point by point; for free electron lasers, a large number of differently oriented images of the same sample are combined into a three-dimensional image.
single exposure three-dimensional imaging remains a common goal sought in many imaging methods that have certain difficulties in achieving single exposure three-dimensional imaging. It is known that when an electromagnetic wave passes through a scattering sample, the scattering signal in the far field is the modulation of the amplitude on the EWald sphere by the scattering potential. When the angle covered by the scatter signal is small, the signal detected on the planar CCD can be approximated as a projection on the Ewald ball. To obtain the three-dimensional structure of the sample, the three-dimensional structure of the Fourier space needs to be measured firstly, and the three-dimensional measurement of the Fourier space can be realized by two modes, one mode is to measure the EWald balls with different angles, and the other mode is to measure the EWald balls with different energies. In CT imaging, the equal angle or equal slope measurement can be carried out on a sample, and hundreds or tens of angles are required; recently developed single-orientation three-dimensional coherent diffraction imaging is to scan fourier space by measuring different energies of the Ewald sphere. Compared with the two modes, the latter is more favorable for realizing dynamic three-dimensional imaging.
in a visible laser coherent diffraction imaging experiment, because a high-angle diffraction signal reflects the structure and the property of a sample, a central diffraction spot plays a key role in image reconstruction, and generally direct light needs to be shielded by a baffle, which can simultaneously shield a part of diffraction signals, so that how to simultaneously obtain the high-angle and central diffraction spots and improve the three-dimensional reconstruction precision is a technical problem which needs to be urgently solved by technical personnel in the field.
Disclosure of Invention
in order to overcome the defects of the prior art, the invention provides a single-exposure high-numerical-aperture laser coherent diffraction imaging device and a using method thereof, wherein the device adopts a combined reflecting plane mirror to reflect a high-angle diffraction signal, and a high-sensitivity liquid nitrogen cooled CCD image sensor is used for recording; the device can obtain a high-numerical aperture diffraction image of a sample through single exposure by adopting a reflecting plane mirror with a central hole to reflect a low-angle diffraction signal and recording the signal by using a common CCD image sensor. The three-dimensional structure and the properties of the sample are analyzed by the angle-resolved diffraction signal of the sample, and the method has important application value in materials science and biology.
In order to achieve the purpose, the invention adopts the following technical scheme:
A single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device is sequentially and coaxially provided with a pulse laser (1), an attenuation sheet (2), a polarizer (3), a first lens (4), a diaphragm (5), a sample table (6), a reflector group (7), a reflector with a hole (8) and a light beam collector (9) along the advancing direction of a light beam; a second lens (10) and a liquid nitrogen cooling CCD image sensor (11) are sequentially arranged along the advancing direction of the reflected light beam of the reflector group (7); and a common CCD image sensor (12) is arranged in the forward direction of a reflected light beam of the reflector (8) with the hole, and the liquid nitrogen cooling CCD image sensor (11) and the common CCD image sensor (12) are both connected with a computer (13).
furthermore, the reflector group (7), the reflector with the hole (8) and the two CCD image sensors (11 and 12) are arranged on the stepping frame and can move left and right, up and down and back and forth.
Furthermore, the reflector group (7) is composed of a plurality of coplanar plane mirrors separated from each other, and light beams reach the perforated reflector (8) through gaps of the reflector group.
Further, a diaphragm is placed at the focal point of the lens near the light source.
Further, a sample stage was placed at the laser's waist spot.
further, the number of pixels of the liquid nitrogen cooled CCD was 1300 × 1340, and the pixel size was 20 μm; the number of pixels of a common CCD is 656 × 492, and the pixel size is 5.6 μm.
according to another object of the present invention, the present invention further provides a method for using a single-exposure high-na pulsed laser coherent diffraction imaging device, comprising the following steps:
The first step is as follows: opening the pulse laser, and adjusting the position of each device in the light path to enable the pulse laser to pass through the light path;
The second step is that: fixing a sample on a sample table, and adjusting the position of the sample table to enable pulse laser to directly irradiate the sample; adjusting the position of a reflector group (7) to symmetrically project diffraction signals onto a liquid nitrogen cooled CCD image sensor (11); the position of the reflector (8) with the hole is adjusted to enable a central diffraction light spot to appear on the common CCD; controlling the shutter so that the two CCD image sensors (11, 12) are exposed simultaneously;
The third step: storing the diffraction signals to a computer; the computer reconstructs a three-dimensional image of the sample according to the diffraction signal;
The fourth step: opening a sample injector to enable a large number of same samples to enter an optical path, recording diffraction signals of the samples, and synthesizing the diffraction signals into a three-dimensional image of the samples;
The fifth step: and (3) irradiating the sample by using a pulse laser, recording diffraction images of the sample at different moments, and reconstructing a three-dimensional dynamic image of the sample.
Further, the adjusting the position of each device in the optical path in the first step includes: the direction of the polarizer (3) is adjusted to ensure that the laser polarization plane is in the vertical direction; adjusting the distance between a plurality of plane mirrors in the reflector group (7), and adjusting the reflector group (7) in the direction perpendicular to the light path to allow direct pulse laser to pass through; adjusting a perforated reflector 8 in a direction perpendicular to the optical path to enable the pulse laser to pass through the round hole; and adjusting the beam collector in the direction vertical to the light path to enable the pulse laser to enter the beam collector.
Furthermore, the attenuation times of the attenuation sheets are adjusted in the second step, so that diffraction signals on the liquid nitrogen cooling CCD image sensor (11) are close to saturation.
further, the method for reconstructing the three-dimensional image of the sample in the third step comprises the following steps: the low-angle diffraction signal collected by the common CCD image sensor (12) is used for filling a central diffraction signal of a high-angle diffraction image collected by the liquid nitrogen cooling CCD image sensor (11), after the whole image is distorted and corrected, an iterative algorithm is used for obtaining a plane amplitude and phase image of a reconstructed object, and the diffraction image is projected to an Ewald ball to reconstruct a three-dimensional image of a sample.
The invention has the advantages of
1. According to the single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device, the polarizer is added, so that the accuracy of a high-angle diffraction signal in reconstruction is improved;
2. the invention utilizes a single lens and an attenuation sheet to compress light beams, thereby simplifying light paths while ensuring the luminous flux;
3. The invention uses a reflector group to reflect high-angle diffraction signals, and uses a lens and a liquid nitrogen cooled CCD sensor to record diffraction images; the diffraction signal is recorded by a common CCD image sensor, the high-angle and low-angle diffraction signals are recorded simultaneously, the high-angle diffraction signal is supplemented by the low-angle diffraction signal to obtain a single-exposure high-numerical-aperture diffraction image, the diffraction image can be used for reconstructing a plane image of a sample through distortion correction, and the three-dimensional image of the sample can be reconstructed by projecting the diffraction image on an EWald ball.
4. The invention uses the sample injector to make a large number of same samples enter the light path, and can record the diffraction images of the samples and reconstruct the three-dimensional images of the samples by using a mathematical algorithm. By illuminating the sample with a pulsed laser, a three-dimensional dynamic image of the sample can be recorded. Has important application value for analyzing the dynamic three-dimensional structure and properties of micron-sized materials and animal and plant cells.
drawings
The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the application and, together with the description, serve to explain the application and are not intended to limit the application.
FIG. 1 is a schematic diagram of a single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device.
Wherein: the device comprises a pulse laser 1, an attenuation sheet 2, a polarizer 3, a lens 4, a diaphragm 5, a sample stage 6, a reflector group 7, a reflector 8 with a hole, a light beam collector 9, a lens 10, a liquid nitrogen cooled CCD image sensor 11, a common CCD image sensor 12 and a computer 13.
FIG. 2 is a sample of coherent diffraction imaging, a graph of microsphere silica gel alignment.
Fig. 3 is the experimental results according to examples 1 and 2.
FIG. 4 is an image of a material object reconstructed using an oversampling and iterative algorithm.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
the embodiments and features of the embodiments in the present application may be combined with each other without conflict.
Example one
The embodiment discloses a single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device, as shown in fig. 1.
a pulse laser 1, an attenuation sheet 2, a polarizer 3, a lens 4, a diaphragm 5, a sample stage 6, a reflector group 7, a reflector with holes 8 and a light beam collector 9 are coaxially arranged along the advancing direction of a light beam in sequence; a lens 10 and a liquid nitrogen cooling CCD image sensor 11 are sequentially arranged along the advancing direction of the reflected light beam of the reflector group 7; and a common CCD image sensor 12 is arranged in the forward direction of a reflected light beam of the reflector 8 with the hole, and the liquid nitrogen cooling CCD image sensor 11 and the common CCD image sensor 12 are both connected with a computer 13.
The reflector group 7 and the reflector with holes 8 are arranged on a stepping frame which can move left and right, up and down and back and forth; the attenuation sheet 2, the polarizer 3, the diaphragm 5 and the light beam collector 9 are fixed on an optical bench, and the two CCDs are respectively fixedly arranged on two mutually vertical stepping frames capable of moving left and right and up and down and then fixed on the stepping frames capable of moving back and forth along the direction of the light path.
The mirror group 7 consists of a number of plane mirrors at a distance from each other so that the light beam can reach the perforated mirror 8 through the gaps of the mirror group. Preferably, the reflector group 7 is composed of four coplanar flat mirrors, and the distances between the four flat mirrors can be adjusted. The distance between the plane mirror group and the sample can be adjusted along the light path direction; the reflector with the hole can adjust the distance to the reflecting plane mirror group along the light path direction. The lens 10 and the liquid nitrogen cooled CCD image sensor are placed on one side of the reflecting plane mirror group, the distance between the lens and the reflecting plane mirror can be adjusted along the stepping frame, and the distance between the lens and the reflecting plane mirror can be adjusted along the stepping frame by placing the common CCD on one side of the plane mirror with the hole.
In the above single exposure high numerical aperture pulsed laser coherent diffraction imaging device: the pulse laser is a picosecond laser, and the wavelength of output light is 0.532 micron; the attenuation sheet attenuates the laser emitted from the laser, so that the diffraction signal of the CCD11 is not saturated, and the attenuation multiple can be selected to be 50 or 100; the focal length of the lens 4 is 500mm and is used for compressing light beams, and the focal length of the lens 10 is 300mm and is used for converging high-angle diffraction light spots; the diaphragm is arranged at the position of the focal point of the lens biased to the light source and used for eliminating stray light of a light path to obtain a purified light beam; the plane reflector group consists of four plane reflectors, and the side length is 2 cm-4 cm, preferably 2.5 cm; the plane reflector with the holes is a square plane reflector, and the side length is 2 cm-4 cm, preferably 3 cm. The sample stage was placed at the laser waist spot. The pixel number of the liquid nitrogen cooled CCD is 1300 multiplied by 1340, and the pixel size is 20 micrometers; the number of pixels of a common CCD is 656 × 492, and the pixel size is 5.6 μm.
Based on the imaging device, light beams are reflected by a reflector group 7 and then spread into a lens 10, and then projected to a liquid nitrogen cooled CCD image sensor 11; the light beam is reflected by the reflector 8 with the hole and then projected to the common CCD image sensor 12, and the liquid nitrogen-cooled CCD image sensor 11 and the common CCD image sensor 12 send collected diffraction signals to the computer 13 for reconstructing three-dimensional imaging.
Example two
According to the device in the first embodiment, the embodiment provides a using method of the device.
A use method of a single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device comprises the following steps:
The first step is as follows: opening a pulse laser, collecting signals by using a CCD (charge coupled device), and adjusting the position of each device in a light path to enable the pulse laser to pass through the light path;
Specifically, the direction of the polarizer 3 is adjusted to make the laser polarization plane in the vertical direction; adjusting the distance between four plane mirrors in the reflector group 7, and adjusting the reflector group 7 to allow direct pulse laser to pass through in the direction perpendicular to the light path; adjusting a perforated reflector 8 in a direction perpendicular to the optical path to enable the pulse laser to pass through the round hole; adjusting the beam collector in the direction perpendicular to the light path to enable the pulse laser to enter the beam collector;
The second step is that: fixing a sample on a sample table, and adjusting the position of the sample table to enable pulse laser to directly irradiate the sample; adjusting the attenuation times of the attenuation sheets to enable diffraction signals on the liquid nitrogen cooling CCD to be close to saturation; adjusting the reflector group 7 to be 2-3 cm away from the sample along the light path, and adjusting the planar reflector group 7 in the direction vertical to the light path to ensure that diffraction signals are symmetrically distributed on the four plane mirrors; adjusting the plane mirror to ensure that the diffraction signals are symmetrically distributed on the CCD11, and adjusting the distance from the lens to the CCD to collect high-angle diffraction signals; adjusting the distance from the plane mirror with the hole to the sample along the light path to enable a central diffraction light spot to appear on the common CCD; controlling the shutter to expose the two CCDs simultaneously;
The third step: storing the diffraction signals to a computer; the computer reconstructs a three-dimensional image of the sample according to the diffraction signal;
Specifically, the low-angle diffraction signal collected by the common CCD image sensor 12 is used for filling a central diffraction signal of a high-angle diffraction image collected by the liquid nitrogen cooling CCD image sensor 11, the whole image is distorted and corrected, an iterative algorithm is used for obtaining a plane amplitude and phase image of a reconstructed object, and the diffraction image is projected onto an EWald ball to reconstruct a three-dimensional image of a sample.
The fourth step: opening a sample injector to enable the same samples to enter a light path in sequence, recording diffraction signals of the samples, and synthesizing the diffraction signals into a three-dimensional image of the sample;
The fifth step: and (3) irradiating the sample by using a pulse laser, recording diffraction images of the sample at different moments, and reconstructing a three-dimensional dynamic image of the sample.
The invention improves the accuracy of the high-angle diffraction signal in reconstruction by introducing the polarizer; the light beams are compressed by using a single lens and an attenuation sheet, so that the light path is simplified while the light flux is ensured; the reflecting light of the reflector group and the reflector with holes is adopted and recorded by the liquid nitrogen-cooled CCD sensor and the common CCD image sensor respectively, so that the simultaneous recording of high-angle diffraction signals and low-angle diffraction signals is realized, the high-angle diffraction signals are supplemented by the low-angle diffraction signals to obtain single-exposure high-numerical-aperture diffraction images, the reconstruction precision of three-dimensional images is provided, and the method has important application value for analyzing the dynamic three-dimensional structures and properties of micron-sized materials and animal and plant cells.
Although the embodiments of the present invention have been described with reference to the accompanying drawings, it is not intended to limit the scope of the present invention, and it should be understood by those skilled in the art that various modifications and variations can be made without inventive efforts by those skilled in the art based on the technical solution of the present invention.
Claims (9)
1. A single-exposure high-numerical-aperture pulse laser coherent diffraction imaging device is characterized in that a pulse laser (1), an attenuation sheet (2), a polarizer (3), a first lens (4), a diaphragm (5), a sample stage (6), a reflector group (7), a reflector with a hole (8) and a light beam collector (9) are coaxially arranged in sequence along the advancing direction of a light beam; a second lens (10) and a liquid nitrogen cooling CCD image sensor (11) are sequentially arranged along the advancing direction of the reflected light beam of the reflector group (7); a common CCD image sensor (12) is arranged in the forward direction of a reflected light beam of the perforated reflector (8), the liquid nitrogen cooled CCD image sensor (11) and the common CCD image sensor (12) are both connected with a computer (13), the reflector group (7) is composed of a plurality of coplanar plane mirrors separated from each other, and the light beam reaches the perforated reflector (8) through the gap of the reflector group (7).
2. the single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device according to claim 1, wherein the mirror group (7), the mirror with the hole (8) and the two CCD image sensors (11, 12) are arranged on a stepping frame capable of moving left and right, up and down, and back and forth.
3. The single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging apparatus according to claim 1, wherein the stop is placed at a position where the focal point of the lens is close to the light source.
4. The single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging apparatus according to claim 1, wherein the sample stage is placed at a waist spot of the laser.
5. The single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device according to claim 1, wherein the number of pixels of the liquid nitrogen cooled CCD is 1300 x 1340, and the pixel size is 20 μm; the number of pixels of a common CCD is 656 × 492, and the pixel size is 5.6 μm.
6. A method of using a single-exposure high-na pulsed laser coherent diffraction imaging device according to any one of claims 1 to 5, comprising the steps of:
The first step is as follows: opening the pulse laser, and adjusting the position of each device in the light path to enable the pulse laser to pass through the light path;
the second step is that: fixing a sample on a sample table, and adjusting the position of the sample table to enable pulse laser to directly irradiate the sample; adjusting the position of a reflector group (7) to ensure that diffraction signals projected onto a liquid nitrogen cooling CCD image sensor (11) are symmetrical; the position of the reflector (8) with the hole is adjusted to enable a central diffraction light spot to appear on the common CCD image sensor; controlling the shutter so that the two CCD image sensors (11, 12) are exposed simultaneously;
the third step: storing the diffraction signals to a computer; the computer reconstructs a three-dimensional image of the sample according to the diffraction signal;
The fourth step: opening a sample injector to enable a large number of same samples to enter an optical path, recording diffraction signals of the samples, and synthesizing the diffraction signals into a three-dimensional image of the samples;
the fifth step: and (3) irradiating the sample by using a pulse laser, recording diffraction images of the sample at different moments, and reconstructing a three-dimensional dynamic image of the sample.
7. The method for using the single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device according to claim 6, wherein the adjusting the position of each device in the optical path in the first step comprises: the direction of the polarizer (3) is adjusted to ensure that the laser polarization plane is in the vertical direction; adjusting the distance between a plurality of plane mirrors in the reflector group (7), and adjusting the reflector group (7) in the direction perpendicular to the light path to allow direct pulse laser to pass through; adjusting a reflector (8) with a hole in the direction vertical to the light path to enable the pulse laser to pass through the round hole; and adjusting the beam collector in the direction vertical to the light path to enable the pulse laser to enter the beam collector.
8. the use method of the single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device according to claim 6, wherein the second step is further to adjust the attenuation multiple of the attenuation sheet so that the diffraction signal on the liquid nitrogen cooled CCD image sensor (11) is close to saturation.
9. The method for using the single-exposure high-numerical-aperture pulsed laser coherent diffraction imaging device according to claim 6, wherein the method for reconstructing the three-dimensional image of the sample in the third step is as follows: the low-angle diffraction signal collected by the common CCD image sensor (12) is used for filling a central diffraction signal of a high-angle diffraction image collected by the liquid nitrogen cooling CCD image sensor (11), after the whole image is distorted and corrected, an iterative algorithm is used for obtaining a reconstructed object plane amplitude and phase image, and the diffraction image is projected onto an Everdet diffraction ball to reconstruct a three-dimensional image of a sample.
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| CN102981261A (en) * | 2012-11-30 | 2013-03-20 | 山东大学 | Laser coherence diffraction microscopic imaging device and application thereof |
| CN104536125A (en) * | 2015-01-13 | 2015-04-22 | 山东大学 | Longitudinal-separated-reference-added multi-cascade scanning coherent diffraction microscopy imaging device and application |
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| CN102981261A (en) * | 2012-11-30 | 2013-03-20 | 山东大学 | Laser coherence diffraction microscopic imaging device and application thereof |
| CN102981261B (en) * | 2012-11-30 | 2015-03-11 | 山东大学 | Laser coherence diffraction microscopic imaging device and application thereof |
| CN104536125A (en) * | 2015-01-13 | 2015-04-22 | 山东大学 | Longitudinal-separated-reference-added multi-cascade scanning coherent diffraction microscopy imaging device and application |
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