CN110801206B - Image correction method of high-resolution optical coherence confocal microscope - Google Patents
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Abstract
The invention discloses an image correction method of a high-resolution optical coherence confocal microscope, which can solve the problems of curvature artifacts caused by position offset of a coherence gate and fuzzy deformation of a view field edge due to the use of a high numerical aperture lens in an optical coherence confocal microscope system; the technical scheme includes that a calibration phase mapping chart is established by analyzing and restoring phase information of a reference surface, and the chart is mapped into an original image, so that the curvature of a coherence gate is corrected, the phase information can be directly used for coherence processing of an OCM image, and system dispersion of any order can be corrected and eliminated in a calculation mode, and imaging performance is improved on the basis of not changing a system light path structure.
Description
Technical Field
The invention relates to an image correction method, in particular to an image correction method of a high-resolution optical coherence confocal microscope.
Background
In the field of medical imaging, OCT (optical coherence tomography) is a novel optical imaging technique that helps medical staff to observe the structure and morphology of the tissue interior and to find microscopic lesions. Based on the principle of light interference, infrared light is used as a light source, the infrared light can penetrate a certain depth of a scanning medium, a detection signal is restored into a longitudinal sectional view of a tissue through calculation, and the infrared light biological tissue detection system has the advantages of high resolution, non-contact and no mark, and can reflect important information such as the internal structure, the scattering coefficient and the like of a biological tissue as a judgment means of in-vivo pathology. In recent years, multi-mode imaging systems that combine OCT technology with other imaging technologies and provide more accurate diagnostic basis have become a focus of research.
Generally, the magnification of the OCT system is limited by the field lens, the lateral resolution is not high, and the longitudinal resolution decreases with the change of the imaging depth, so that it is not easy to balance the imaging depth and the resolution. The confocal microscopy (LSCM) can overcome the above-mentioned problems, and under a microscope objective with a high numerical aperture, a higher lateral resolution is achieved by the conjugate relationship between the illumination pinhole and the detection pinhole and the irradiated point or the detected point, but the depth of confocal scanning imaging is shallow, and a fluorescent agent is needed to perform dyeing imaging on a sample, so that the application range is limited, and thus, the two imaging methods have advantages and disadvantages. Patent CN 107328743 a proposes an optical coherence confocal microscopy system, i.e. an OCM endoscopy system (optical coherence microscopy), which combines the advantages of OCT and confocal imaging means by designing and improving an optical system, so as to significantly improve the OCT resolution without significantly reducing the imaging depth, and apply it to an endoscope.
However, in an optical coherence confocal microscopy system, due to the characteristics of an optical fiber and the use of a high numerical aperture lens, the position shift and curvature change of a coherence gate in an optical scanning process become obvious, particularly some non-telecentric lenses, a back focal plane of a scanning lens does not coincide with a central axis of a scanning mechanism, and scanning of a light beam in the system can cause the change of an optical path, so that the position of the coherence gate of an OCM (optical coherence tomography) is also shifted, curvature artifacts occur, the image intensity and resolution are not uniform, the edge of an imaging field of view can be blurred and deformed, and even images disappear. Furthermore, the telecentric lens is often composed of a plurality of lens groups, the optical design is complex, perfect alignment is difficult to achieve while miniaturization is impossible, and the optical path change is eliminated. Thus limiting the imaging field of view of existing systems and also affecting the imaging performance.
Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
The invention aims to provide an image correction method of a high-resolution optical coherence confocal microscope, and aims to solve the problems of curvature artifacts caused by position deviation of a coherence gate and blurred deformation of a view field edge due to the use of a high numerical aperture lens in an optical coherence confocal microscope system.
The technical scheme of the invention is as follows: an image correction method of high-resolution optical coherence confocal microscopy comprises the following steps:
placing a plane mirror on a sample arm of an OCM system to obtain the two-dimensional coherence gate position curvature of an OCM signal of a mirror surface of the plane mirror, wherein the OCM signal is a depth detection signal of one point in space and is represented as A (x, y, k), x and y are space dimensions, k is a light source wave number, and the two-dimensional coherence gate position curvature is phase distribution of a coherence gate;
calculating a calibration phase mapping chart capable of smoothing the curvature according to the phase distribution of the coherent gate;
apodizing the mirror surface signal of the original plane reflector through a proper window function and carrying out waveform shaping;
subtracting background noise from the windowed and waveform-shaped signal to obtain an original OCM signal after preliminary processing;
multiplying the original interference spectrum by the corresponding calibration phase in the calibration phase mapping chart of each pixel of the original OCM signal after the preliminary processing;
and summing the wave numbers of the signals multiplied by the calibration phase to obtain an optimized OCM image.
The image correction method of the high-resolution optical coherence confocal microscope comprises the steps of placing a plane reflector on a sample arm of an OCM system, aligning the plane reflector with a light beam focus to collect back scattered light, and calculating an OCM signal of a mirror surface of the plane reflector to measure the position curvature of a two-dimensional coherence gate.
The image correction method of the high-resolution optical coherence confocal microscope comprises the steps of placing a plane reflector on a sample arm of an OCM system, and obtaining two-dimensional coherence gate position curvature of an OCM signal of a mirror surface of the plane reflector, wherein the OCM signal is a depth detection signal of one point in space and is represented as A (x, y, k), x and y are space dimensions, and k is a light source wave number.
The image correction method of the high-resolution optical coherence confocal microscope comprises the following steps of obtaining phase distribution of a coherence gate according to the position curvature of the coherence gate of a plane mirror surface signal, wherein the phase distribution is the phase distribution of the position of the whole coherence gate, and the phase distribution of the coherence gate is obtained by the following method: by collecting a plurality of OCM signal graphs with different phases, the coherent gate phase of the reconstructed image only containing the object information is obtained through multi-step phase shift calculation.
The image correction method of the high-resolution optical coherence confocal microscope comprises the following steps of obtaining a coherence gate phase of a reconstructed image only containing object information through four-step phase shift calculation, wherein a calculation formula is as follows:
wherein,
the phase of the coherent gate is represented,
、
、
、
respectively representing the OCM signal strength in 4 different phase states.
The image correction method of the high-resolution optical coherence confocal microscope includes the steps of calculating a calibration phase mapping chart capable of flattening curvature according to phase distribution of a coherence gate, wherein the calibration phase mapping chart can be obtained by the following formula 2:
wherein,
denotes the calibration phase map and C (x, y) is the corrected surface coefficients.
The image correction method of the high-resolution optical coherence confocal microscope comprises the following steps of apodizing a mirror surface signal of an original plane reflector through a proper window function and carrying out waveform shaping, wherein the proper window function is a Hanning window, and the window expression is w (k):
wherein,
is the wavenumber spacing.
The image correction method of the high-resolution optical coherence confocal microscope is characterized in that the signals after the waveform shaping processing are B (x, y, k):
The image correction method of the high-resolution optical coherence confocal microscope comprises the following steps of subtracting background noise from a windowed and waveform-shaped signal to obtain an original OCM signal subjected to primary processing, wherein the OCM signal subjected to primary processing is a signal C (x, y, k) obtained by subtracting the background noise:
where N (x, y) is the system background noise distribution.
The image correction method of the high-resolution optical coherence confocal microscope comprises the steps of multiplying an original interference spectrum by a corresponding calibration phase in a calibration phase mapping chart for each pixel of an original OCM signal, summing the signal multiplied by the calibration phase by wave number to obtain an optimized OCM image, wherein the OCM signal multiplied by the corresponding calibration phase and the wave number k are summed by the following formula 6:
wherein z is the distance between the imaging plane and the focal plane, and z = 0.
The invention has the beneficial effects that: the invention provides an image correction method of a high-resolution optical coherence confocal microscope, which can solve the problems of curvature artifacts caused by position offset of a coherence gate and fuzzy deformation of a view field edge due to the use of a high numerical aperture lens in an optical coherence confocal microscope system; the technical scheme includes that a calibration phase mapping chart is established by analyzing and restoring phase information of a reference surface, and the chart is mapped into an original image, so that the curvature of a coherence gate is corrected, the phase information can be directly used for coherence processing of an OCM image, and system dispersion of any order can be corrected and eliminated in a calculation mode, and imaging performance is improved on the basis of not changing a system light path structure.
Drawings
Fig. 1 is a schematic diagram of a simplified high numerical aperture non-telecentric system of the present invention.
Fig. 2 is a flowchart of the steps of the image correction method of the high-resolution optical coherence confocal microscope of the present invention.
Fig. 3a is a graph of the original OCM at z =0 in the present invention.
Fig. 3b is the original OCM at z =20um in the present invention.
FIG. 3c is a diagram of the OCM processed by the calibration method of the present invention.
FIG. 3d is a diagram of a corrected phase distribution according to the present invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like, indicate orientations and positional relationships based on those shown in the drawings, and are used only for convenience of description and simplicity of description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be considered as limiting the present invention. Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, features defined as "first", "second", may explicitly or implicitly include one or more of the described features. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; may be mechanically connected, may be electrically connected or may be in communication with each other; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless otherwise expressly stated or limited, "above" or "below" a first feature means that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact with each other via another feature therebetween. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly under and obliquely below the second feature, or simply meaning that the first feature is at a lesser elevation than the second feature.
The following disclosure provides many different embodiments or examples for implementing different features of the invention. To simplify the disclosure of the present invention, the components and arrangements of specific examples are described below. Of course, they are merely examples and are not intended to limit the present invention. Furthermore, the present invention may repeat reference numerals and/or letters in the various examples, such repetition is for the purpose of simplicity and clarity and does not in itself dictate a relationship between the various embodiments and/or configurations discussed. In addition, the present invention provides examples of various specific processes and materials, but one of ordinary skill in the art may recognize applications of other processes and/or uses of other materials.
As shown in fig. 1, in a simplified high-na non-telecentric system, when a light beam passes through an input diaphragm by means of scanning or rotation of a micro-galvanometer, the optical path lengths of the light beam from different positions to a focal plane are changed, generally, the optical path length at positions beyond the edge is shortened, and as a result of two-dimensional scanning, a curved coherence gate is formed, and the positions of the coherence gates at equal optical paths are distributed in a circular shape, so that the optical paths are not equal in the same plane. In an imaging system, a real high numerical aperture optical system is much more complex than the above situation, and is influenced by factors such as processing technology and assembly technology, and the curvature distribution of a focal plane of the system is not completely a circle, but shows non-uniform and non-linear distribution.
As shown in fig. 2, an image correction method of high-resolution optical coherence confocal microscopy based on a high-numerical-aperture non-telecentric system includes the following steps:
s1: placing a plane mirror on a sample arm of an Optical Coherence Microscopy (OCM) system to obtain a two-dimensional coherence gate position curvature of an OCM signal of a plane mirror surface of the plane mirror, wherein the OCM signal is a depth detection signal of one point in space and is represented as A (x, y, k), x and y are space dimensions, k is a light source wave number, and the two-dimensional coherence gate position curvature is phase distribution of a coherence gate;
s2: calculating a calibration phase mapping chart capable of smoothing the curvature according to the phase distribution of the coherent gate;
s3: apodizing the mirror surface signal of the original plane reflector through a proper window function and carrying out waveform shaping;
s4: subtracting background noise from the windowed and waveform-shaped signal to obtain an original OCM signal after preliminary processing;
s5: multiplying the original interference spectrum by the corresponding calibration phase in the calibration phase mapping chart by each pixel of the original OCM signal;
s6: and summing the wave numbers of the signals multiplied by the calibration phase to obtain an optimized OCM image.
Specifically, in step S1, the plane mirror is aligned to the beam focus to collect the backscattered light to the maximum, and the position curvature of the two-dimensional coherence gate can be measured by calculating the OCM signal of the plane mirror.
Specifically, in step S1, the OCM signal is a depth detection signal at a certain point in space, which can be represented as a (x, y, k), where x and y are spatial dimensions, and k is a light source wave number.
Specifically, in step S1, the phase distribution is the phase distribution of the entire coherent gate position, and the phase distribution of the coherent gate is obtained by: and calculating the phase of a coherent gate of a reconstructed image only containing object information by acquiring a plurality of OCM signal graphs with different phases. The phase of the coherent gate for calculating the reconstructed image containing only the object information can be calculated by multi-step phase shift (e.g., three-step phase shift, four-step phase shift, five-step phase shift). In this embodiment, the coherent gate phase of the reconstructed image containing only the object information is obtained through four-step phase shift calculation, and the calculation formula is as follows:
wherein,
the phase of the coherent gate is represented,
、
、
、
respectively representing the OCM signal strength in 4 different phase states.
Specifically, in step S2, the calibration phase map table may be obtained by equation 2:
wherein,
denotes the calibration phase map, and C (x, y) is the corrected surface coefficient.
Specifically, in step S3, due to the coherence of the light source, after the OCM signal is subtracted from the background noise, the OCM signal has strong side lobes after hilbert transform, and it is a common practice to apodize the interference signal with an appropriate window function and shape the waveform before hilbert transform, so as to suppress interference terms due to the side lobes and achieve better image contrast. Wherein, the suitable window function is generally a hanning window commonly used in signal processing, and the window expression is w (k):
wherein,
is the wavenumber spacing;
the waveform shaping includes but is not limited to processing modes such as signal normalization and low-frequency direct current term removal, and the signals after the waveform shaping are B (x, y, k):
Specifically, in step S4, the OCM signal after the preliminary processing is the signal C (x, y, k) obtained by subtracting the background noise:
where N (x, y) is the system background noise distribution.
In step S6, the wave number is summed with the signal multiplied by the calibration phase to obtain a corrected OCM image, the curvature of the coherence gate is corrected, the dispersion is optimally compensated, and an optimized OCM image with an unlimited viewing field is obtained by combining with necessary image processing (such as image brightness amplification, median filtering, etc.).
Specifically, in step S5 and step S6, each pixel of the optimized OCM image with unlimited field of view is obtained by multiplying the original OCM signal by the corresponding calibration phase table and summing the wave numbers k, which is obtained by equation 6:
wherein z is the distance between the imaging plane and the focal plane. As a preferred embodiment, z is zero, which means that the imaging plane is just at the system focal plane, and the imaging effect is best.
According to the technical scheme, the image correction method can effectively reduce the complexity of image single-frame processing in the optical coherent confocal microscope system, and in addition, the corrected phase section provides more accurate coherent gate correction than a peak value searching method; in combination with an FPGA (field Programmable Gate array) operation module of the imaging System and a Micro-Electro-Mechanical System (MEMS) galvanometer scanning, the System can still maintain a higher imaging speed.
In which, fig. 3a to 3d show the application effect of the image correction method in the coherent confocal microscopic image of colon tissue. Wherein, fig. 3a is the original OCM image at z =0, and fig. 3b is the original OCM image at z =20um, it can be seen that, the uncorrected OCM image has different imaging definitions at different positions of the field of view, and the out-of-focus position of a partial region is rather clearer than the focal plane image, and there is an error. Fig. 3c is a diagram of the OCM after being processed by the calibration method, and fig. 3d is a diagram of the calibration phase distribution used.
The technical scheme can solve the problems of curvature artifacts caused by the position deviation of a coherence gate and fuzzy deformation of the field of view edge due to the use of a high numerical aperture lens in an optical coherence confocal microscopy system. The technical scheme includes that a calibration phase mapping chart is established by analyzing and restoring phase information of a reference surface, and the chart is mapped into an original image, so that the curvature of a coherence gate is corrected, the phase information can be directly used for coherence processing of an OCM image, and system dispersion of any order can be corrected and eliminated in a calculation mode, and imaging performance is improved on the basis of not changing a system light path structure.
In the description herein, references to the description of the terms "one embodiment," "certain embodiments," "an illustrative embodiment," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. An image correction method of a high-resolution optical coherence confocal microscope is characterized by comprising the following steps:
placing a plane mirror on a sample arm of an OCM system to obtain the two-dimensional coherence gate position curvature of an OCM signal of a mirror surface of the plane mirror, wherein the OCM signal is a depth detection signal of one point in space and is represented as A (x, y, k), x and y are space dimensions, and k is a light source wave number;
obtaining the phase distribution of a coherent gate according to the position curvature of the coherent gate of a plane mirror surface signal;
calculating a calibration phase mapping chart capable of smoothing the curvature according to the phase distribution of the coherent gate;
apodizing the mirror surface signal of the original plane reflector through a proper window function and carrying out waveform shaping;
subtracting background noise from the windowed and waveform-shaped signal to obtain an original OCM signal after preliminary processing;
multiplying the original interference spectrum by the corresponding calibration phase in the calibration phase mapping chart of each pixel of the original OCM signal after the preliminary processing;
and summing the wave numbers of the signals multiplied by the calibration phase to obtain an optimized OCM image.
2. The method of claim 1, wherein a plane mirror is placed on the sample arm of the OCM system, the plane mirror is aligned to the beam focus to collect the backscattered light, and the curvature of the position of the two-dimensional coherence gate can be measured by calculating the OCM signal of the plane mirror.
3. The method for image correction of high-resolution optical coherence confocal microscopy according to claim 1 or 2, wherein a plane mirror is placed on a sample arm of the OCM system to obtain two-dimensional coherence gate position curvature of the OCM signal of the plane mirror surface.
4. The image correction method of the high-resolution optical coherence confocal microscope according to claim 3, wherein the phase distribution of the coherence gate is obtained according to the curvature of the coherence gate position of the plane mirror image signal, and the phase distribution is the phase distribution of the whole coherence gate position, and the phase distribution of the coherence gate is obtained by: by collecting a plurality of OCM signal graphs with different phases, the coherent gate phase of the reconstructed image only containing the object information is obtained through multi-step phase shift calculation.
5. The image correction method of the high-resolution optical coherence confocal microscope according to claim 4, wherein the phase of the coherence gate of the reconstructed image containing only the object information is obtained by four-step phase shift calculation, and the calculation formula is as follows:
wherein,
the phase of the coherent gate is represented,
、
、
、
respectively representing the OCM signal strength in 4 different phase states.
6. The image correction method for high-resolution optical coherence confocal microscopy according to claim 5, wherein a calibration phase map capable of flattening curvature is calculated from the phase distribution of the coherence gate, and the calibration phase map is obtained by equation 2:
wherein,
denotes the calibration phase map and C (x, y) is the corrected surface coefficients.
7. The image correction method of high-resolution optical coherence confocal microscopy according to claim 6, wherein the original plane mirror surface signal is apodized and waveform shaped by a suitable window function, wherein the suitable window function is Hanning window, and the window expression is w (k):
wherein,
is the wavenumber spacing.
8. The method of claim 7, wherein the waveform-shaped signals are B (x, y, k):
9. The image correction method of the high-resolution optical coherence confocal microscope according to claim 8, wherein the original OCM signal after preliminary processing is obtained by subtracting the background noise from the windowed and waveform-shaped signal, and the OCM signal after preliminary processing is a signal C (x, y, k) after subtracting the background noise:
where N (x, y) is the system background noise distribution.
10. The image correction method of high-resolution optical coherence confocal microscopy according to claim 9, wherein the initial interference spectrum is multiplied by the corresponding calibration phase in the calibration phase mapping table for each pixel of the original OCM signal after the initial processing, and the signal multiplied by the calibration phase is summed with the wave number to obtain the optimized OCM image, wherein the OCM signal is multiplied by the corresponding calibration phase and the wave number k is summed by equation 6:
wherein z is the distance between the imaging plane and the focal plane, and z = 0.
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| CN107328743A (en) * | 2017-07-05 | 2017-11-07 | 广东欧谱曼迪科技有限公司 | A kind of optical coherence confocal microscopy endoscope system and implementation method |
| CN108403079A (en) * | 2018-02-26 | 2018-08-17 | 执鼎医疗科技（杭州）有限公司 | A kind of confocal imaging system based on OCT |
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