Microscopic system for structured light vertical illumination

Abstract

The invention belongs to the technical field of microscopic imaging, and discloses a structured light vertical illumination microscopic system, which comprises an LED light source assembly, a light beam shaping assembly, a spatial light modulator, a light source module, a light source module, a light source module, a light source module, a light source module, a light source module and a light source module, the imaging assembly is used for projecting and imaging the light field modulated by the spatial light modulator to the surface of a target object; the beam splitter prism is used for carrying out beam splitting imaging on the incident light and the reflection light of the target object, so that the incident light and the reflection light are imaged to the camera; and the processing module is used for performing coding control on the spatial light modulator and analyzing the image acquired by the camera to obtain a super-resolution image. The LED light and the spatial light modulator are adopted to generate structured light to illuminate a target, data of a shot picture are processed through a structured light illumination demodulation algorithm model, and super-resolution imaging is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of microscopic imaging, and in particular relates to a microscopic system with structured light epi-illumination. Background Art

[0002] Structured illumination microscopy (SIM) is a technology that inserts a structured light generator into the illumination light path to form a pattern with regularly changing brightness, projects it onto the sample, and obtains a super-resolution image through Fourier transform.

[0003] In the process of realizing the present invention, the inventors found that there are at least the following problems in the existing structured light illumination microscopic imaging scheme: the existing structured light microscopic imaging system usually adopts multi-wavelength laser mixing, and the imaging quality will be greatly reduced due to the influence of laser light source speckle noise and temperature drift; the existing structured light microscopic imaging system adopts filters and dichroic mirrors, and this design can only collect fluorescence excitation information, and microscopic imaging cannot be performed in the absence of fluorescence excitation; in the existing epi-illumination microscopic imaging system, image acquisition is limited by the optical diffraction limit of the imaging system, resulting in the inability to collect high-frequency image information.

[0004] The above description of the background technology is only for facilitating an in-depth understanding of the technical solution of the present invention (such as the technical means used, the technical problems solved and the technical effects produced), and should not be regarded as an admission or any form of implication that the information constitutes prior art already known to those skilled in the art. Summary of the invention

[0005] The present invention aims to solve the above technical problems at least to a certain extent. To this end, the present invention aims to provide a microscopy system with structured light epi-illumination.

[0006] The technical solution adopted by the present invention is:

[0007] A microscopic system with structured light epi-illumination, comprising:

[0008] LED light source components,

[0009] A beam shaping component is used to shape the light emitted by the LED light source component into a uniform and collimated illumination beam, and irradiate it onto the spatial light modulator through a projection prism;

[0010] An imaging component, used for projecting the light field modulated by the spatial light modulator onto the surface of the target object;

[0011] A beam splitter prism is used to split the incident light and the reflected optics of the target object into separate beams for imaging to the camera;

[0012] The processing module is used to perform coding control on the spatial light modulator and to analyze the image captured by the camera to obtain a super-resolution image.

[0013] Preferably, the imaging assembly comprises a tube lens and a microscope objective lens, wherein the tube lens is located between the beam splitter prism and the camera, and the microscope objective lens is located between the beam splitter prism and the target object. Alternatively, the imaging assembly comprises a telecentric lens.

[0014] Preferably, the LED light source assembly includes a plurality of single-wavelength LED light sources, each of which is provided with a corresponding beam shaping unit, the first beam shaping unit is provided with a corresponding emission prism, and the remaining beam shaping units are provided with a corresponding dichroic mirror, and the emission prism and the dichroic mirror are provided on the same straight line. Alternatively, the LED light source assembly is a white LED.

[0015] Preferably, a barrel mirror is provided between the projection prism and the beam splitting prism.

[0016] Preferably, the processing module analyzes the image captured by the camera through a structured light illumination demodulation algorithm model. The structured light illumination demodulation algorithm model includes:

[0017] Phase analysis module, used to extract phase information of images with different phases, thus providing a basis for subsequent phase correction and reconstruction;

[0018] Phase correction module, used to ensure that spectrum information of different phases and directions can be correctly aligned without phase misalignment;

[0019] The image reconstruction module is used to generate a super-resolution image through spectrum synthesis and inverse Fourier transform, and combine denoising and artifact removal algorithms to obtain the final super-resolution image.

[0020] The beneficial effects of the present invention are:

[0021] The present invention provides a microscopic system of structured light epi-illumination, which uses LED light and a spatial light modulator to generate structured light to illuminate the target, and processes the captured image through a structured light illumination demodulation algorithm model to achieve super-resolution imaging. The present invention can achieve synchronous high-real-time super-resolution imaging of multi-wavelength images; the target can be imaged without fluorescence excitation; the imaging is not affected by laser speckle noise and is not affected by temperature drift. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a schematic diagram of the optical path structure of the microscope system of structured light epi-illumination of the present invention.

[0023] Figure 2 Schematic diagram of the LED light source assembly of the present invention.

[0024] In the figure: 1-LED light source assembly; 11-single wavelength LED light source; 12-beam shaping unit; 13-emitting prism; 14-dichroic mirror; 2-beam shaping assembly; 3-projection prism; 4-spatial light modulator; 5-barrel mirror; 6-beam splitter prism; 71-tube mirror; 72-microscope objective lens; 8-sample; 9-camera. DETAILED DESCRIPTION

[0025] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and are not to be construed as limiting the present invention. The components of the embodiments of the present invention generally described and shown in the drawings herein may be arranged and designed in a variety of different configurations.

[0026] In the present invention, unless otherwise clearly stipulated and limited, terms such as "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, or it can be the internal connection of two elements.

[0027] like Figure 1 and Figure 2 As shown, a microscopy system of structured light incident illumination in this embodiment includes a processing module, an LED light source assembly 1, a beam shaping assembly 2, a projection prism 3, a spatial light modulator 4, a barrel mirror 5, an imaging assembly and a beam splitter prism 6; in this embodiment, the LED light source assembly 1 includes a plurality of single-wavelength LED light sources 11, each single-wavelength LED light source 11 is correspondingly provided with a beam shaping unit 12, the first beam shaping unit 12 is correspondingly provided with an emitting prism 13, and the remaining beam shaping units 12 are correspondingly provided with a dichroic mirror 14, and the emitting prism 13 and the dichroic mirror 14 are arranged on the same straight line, so as to combine LED light sources of different wavelengths to form a multi-wavelength LED light source.

[0028] The beam shaping component 2 is used to shape the light emitted by the LED light source component 1 into a uniform and collimated illumination beam, and irradiate it onto the spatial light modulator 4 through the projection prism 3. The imaging component is used to project the light field modulated by the spatial light modulator 4 onto the surface of the sample 8; the barrel mirror 5 is arranged between the projection prism 3 and the beam splitter prism 6. The beam splitter prism 6 is used to split the incident light and the reflected optics of the sample 8 into beams for imaging, so that it is imaged to the camera 9.

[0029] In this embodiment, the imaging assembly includes a tube lens 71 and a microscope objective lens 72 . The tube lens 71 is located between the beam splitter prism 6 and the camera 9 , and the microscope objective lens 72 is located between the beam splitter prism 6 and the sample 8 .

[0030] The processing module is used to perform encoding control on the spatial light modulator 4 and to analyze the image captured by the camera 9 through a structured light illumination demodulation algorithm model to obtain a super-resolution image.

[0031] The working process of the microscopic system of structured light epi-illumination in this embodiment is as follows:

[0032] Combining the LED light sources of different wavelengths in the LED light source assembly 1 to form a multi-wavelength LED light source;

[0033] shaping the light emitted by the multi-wavelength LED light source into a uniform and collimated illumination beam, and irradiating it onto the spatial light modulator 4;

[0034] Encoding and controlling the spatial light modulator 4 so as to form a desired image;

[0035] The light field modulated by the spatial light modulator 4 is projected onto the surface of the sample 8 by using an imaging component;

[0036] The incident light and the reflected light of the sample 8 are split into beams and imaged by using the beam splitter prism 6 so as to be imaged onto the imaging device surface of the camera 9;

[0037] The captured image is analyzed using the structured light illumination demodulation algorithm model, and the final super-resolution image is obtained through calculation.

[0038] The structured light illumination demodulation algorithm model includes a phase analysis module, a phase correction module, and an image reconstruction module. The phase analysis module is used to extract the phase information of images with different phases, thereby providing a basis for subsequent phase correction and reconstruction; the phase correction module is used to ensure that the spectrum information of different phases and directions can be correctly aligned without phase misalignment, thereby ensuring the accuracy of the final image reconstruction; the image reconstruction module is used to generate super-resolution images through spectrum synthesis and inverse Fourier transform, and combine denoising and artifact removal algorithms to obtain the final super-resolution image.

[0039] The working process of the phase analysis module is:

[0040] Fourier transform: First, perform a two-dimensional Fourier transform on the structured light images of different phases to convert the images from the spatial domain to the frequency domain;

[0041] Spectral peak detection: In the frequency domain, fringe patterns with different phases will produce different spectral shifts; the task of phase resolution is to accurately locate the spectral peak positions of these shifts;

[0042] Phase information extraction: By comparing the spectrum peaks at different phases, the phase information of the fringe pattern can be extracted.

[0043] The working process of the phase correction module is:

[0044] Phase difference calculation: By performing Fourier transform on the fringe patterns at different phases, extracting their spectrum, and calculating the phase difference; the phase difference is caused by the phase change of structured light illumination and needs to be corrected;

[0045] Spectrum translation: Based on the calculated phase difference, the spectrum of each image is translated or rotated in the frequency domain to ensure that their high-frequency components can be aligned in the frequency domain; correct phase correction directly determines the accuracy of subsequent image reconstruction;

[0046] Phase compensation: In order to ensure the accuracy of the spectrum, the phase of each spectrum needs to be compensated so that no phase misalignment occurs during spectrum synthesis.

[0047] The working process of the image reconstruction module is:

[0048] Spectrum synthesis: In the frequency domain, spectrum information of different directions and phases has been extracted; after phase correction, all spectra can be superimposed and synthesized; the synthesized spectrum contains not only the low-frequency information of the original image, but also the high-frequency information captured by structured light modulation;

[0049] Inverse Fourier transform: After the spectrum synthesis is completed, the data in the frequency domain is converted back to the spatial domain using a two-dimensional inverse Fourier transform (IFFT) to obtain a reconstructed image; at this time, the reconstructed image contains a higher spatial resolution than traditional microscopes;

[0050] Artifact removal: Since some artifacts and noise may be introduced during the synthesis and modulation of the spectrum, artifact removal and denoising are performed after image reconstruction. Wiener filtering or regularization methods can be used to smooth the image, retaining effective high-frequency details while reducing unnecessary noise and artifacts.

[0051] The spatial light modulator 4 is a DMD spatial light modulator, an LCOS spatial light modulator or a liquid crystal spatial light modulator.

[0052] It should be noted that, in other embodiments, the imaging assembly includes a telecentric lens. In other embodiments, the LED light source assembly 1 is a white LED.

[0053] The present invention is not limited to the above-mentioned optional implementation modes. Anyone can derive other various forms of products under the inspiration of the present invention. However, no matter what changes are made in the shape or structure, all technical solutions that fall within the scope defined by the claims of the present invention fall within the protection scope of the present invention.

Claims

1. A microscopic system with structured light incident illumination, characterized in that: include: LED light source assembly (1), A beam shaping component (2) is used to shape the light emitted by the LED light source component (1) into a uniform and collimated illumination beam, and irradiate the beam onto the spatial light modulator (4) through a projection prism (3); An imaging component, used for projecting the light field modulated by the spatial light modulator (4) onto the surface of the target object; A beam splitter prism (6) is used to split the incident light and the reflected light of the target object into beams for imaging, so that the images are imaged to the camera (9); The processing module is used to perform coding control on the spatial light modulator (4) and to analyze the image collected by the camera (9) to obtain a super-resolution image.

2. The microscopic system according to claim 1, characterized in that: The imaging component comprises a tube lens (71) and a microscope objective lens (72); the tube lens (71) is located between the beam splitter prism (6) and the camera (9); and the microscope objective lens (72) is located between the beam splitter prism (6) and the target object.

3. The microscopic system according to claim 1, characterized in that: The imaging assembly includes a telecentric lens.

4. The microscopic system according to claim 1, characterized in that: The LED light source assembly (1) comprises a plurality of single-wavelength LED light sources (11), each single-wavelength LED light source (11) being provided with a corresponding beam shaping unit (12), the first beam shaping unit (12) being provided with a corresponding emission prism (13), and the remaining beam shaping units (12) being provided with a corresponding dichroic mirror (14), the emission prism (13) and the dichroic mirror (14) being provided on the same straight line.

5. The microscope system according to claim 1, characterized in that: The LED light source assembly (1) is a white LED.

6. The microscope system according to claim 1, characterized in that: A barrel mirror (5) is provided between the projection prism (3) and the beam splitting prism (6).

7. The microscope system according to claim 1, characterized in that: The processing module analyzes the image collected by the camera (9) through a structured light illumination demodulation algorithm model.

8. The microscopic system according to claim 7, characterized in that: The structured light illumination demodulation algorithm model includes: Phase analysis module, used to extract phase information of images with different phases, thus providing a basis for subsequent phase correction and reconstruction; Phase correction module, used to ensure that spectrum information of different phases and directions can be correctly aligned without phase misalignment; The image reconstruction module is used to generate a super-resolution image through spectrum synthesis and inverse Fourier transform, and combine denoising and artifact removal algorithms to obtain the final super-resolution image.

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