Rapid laser spot scanning control equipment for large imaging view field
By using a combination of a galvanometer scanning mirror and a biaxial microcomputer fast scanning mirror in laser point scanning optical microscopy technology, the problem of slow imaging speed in large imaging fields is solved, and fast and efficient imaging effects are achieved.
Patent Information
- Application Number
- CN202311323831.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-11
- Publication Date
- 2025-04-11
AI Technical Summary
The existing laser point scanning optical microscopy technology has slow imaging speed in large imaging fields and cannot perform phase lock control, making it impossible to achieve fast and efficient imaging.
A pair of galvanometer scanning mirrors are used to perform full-frame scanning and sub-region switching, and a pair of biaxial microcomputer rapid scanning mirrors are used to quickly image sub-regions of interest to avoid imaging areas of uninterested interest.
Fast and efficient imaging in large imaging field is achieved, scanning speed and imaging efficiency are improved, and it is especially suitable for imaging detection of rapidly changing physiological processes.
Smart Images
Figure CN119805734A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of optical microscopic imaging, and in particular to a fast laser point scanning control device for a large imaging field of view. Background Art
[0002] Laser point scanning optical microscopy (such as laser confocal imaging and two-photon microscopy) has the advantages of high spatial resolution and high signal-to-noise ratio, and has become an indispensable research tool in the field of life science and biomedicine. This type of imaging technology mainly relies on scanning mirrors and other equipment for laser point scanning imaging. Existing scanning technologies include: (1) using a pair of galvanometer scanning mirrors for scanning imaging; (2) combining resonant galvanometers and galvanometer scanning mirrors for rapid scanning imaging. Technology (1) has a slow imaging speed; technology (2) has a certain improvement in scanning speed, but the scanning range is small and phase-locked control cannot be performed. Summary of the invention
[0003] In view of this, it is necessary to provide a fast laser point scanning control device that is easy to operate and can be used for a large imaging field of view to address the technical defects of the prior art.
[0004] In order to solve the above problems, the present invention adopts the following technical solutions:
[0005] The present invention discloses a fast laser point scanning control device for a large imaging field of view, characterized in that it comprises a laser control module (100) and a dual-axis microcomputer fast scanning mirror (1), a first lens (2), a second lens (3), an X-axis galvanometer scanning mirror (4), a third lens (5), a fourth lens (6), a Y-axis galvanometer scanning mirror (7), a fifth lens (8), a sixth lens (9), a dichroic mirror (10), an objective lens (11) and a detection module (12) arranged in sequence along the propagation direction of an excitation light beam emitted by the laser control module (100), wherein the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) are used for full-width scanning of the entire imaging field of view and switching between different sub-areas within the imaging field of view, and the dual-axis microcomputer fast scanning mirror (1) is used for fast scanning and imaging of a target position. Wherein:
[0006] The X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) perform full-width scanning imaging on the entire imaging field of view, and select a sub-region of interest. When the dual-axis microcomputer fast scanning mirror (1) is used to scan and image the sub-region of interest, the X / Y galvanometer scanning mirror is in a stationary state: the excitation light beam emitted by the laser control module is reflected by the dual-axis microcomputer fast scanning mirror (1), then transmitted through the first lens (2) and the second lens (3), and then incident on the X-axis galvanometer scanning mirror (4), and then reflected by the X-axis galvanometer scanning mirror (4), transmitted through the third lens (5) and the fourth lens (6), and then incident on the Y-axis galvanometer scanning mirror (7). The galvanometer scanning mirror (7) is incident on the fifth lens (8) after being reflected by the Y-axis galvanometer scanning mirror (7), and then sequentially passes through the sixth lens (9), the dichroic mirror (10) and the objective lens (11) to be focused in the tissue to be tested; the emission light generated after excitation is then transmitted through the objective lens (11), and then is reflected by the dichroic mirror (10) and detected by the detection module (12), thereby completing the scanning imaging of the sub-area; subsequently, the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) switch between different selected sub-areas, and the dual-axis microcomputer fast scanning mirror (1) scans and images the new sub-area; and the scanning imaging of all sub-areas is completed in a cycle.
[0007] In some of these embodiments, the first lens and the second lens constitute an optical relay.
[0008] In some of the embodiments, the third lens and the fourth lens constitute an optical relay.
[0009] The present invention adopts the above technical solution, and its beneficial effects are as follows:
[0010] The present invention uses a pair of galvanometer scanning mirrors to perform full-width scanning within the entire imaging field of view, and thereby determine the imaging sub-region of interest; then the MEMS scanning mirror is used to quickly image the sub-region, and the galvanometer scanning mirror is used to quickly switch between different sub-regions and image them in sequence. This technology avoids imaging uninteresting areas, greatly improves scanning speed and imaging efficiency, and is particularly suitable for imaging and detecting rapidly changing physiological processes in large field imaging. BRIEF DESCRIPTION OF THE DRAWINGS
[0011] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments of the present invention or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0012] Figure 1 A schematic structural diagram of a fast laser point scanning control device for a large imaging field of view provided by the present invention. DETAILED DESCRIPTION
[0013] 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 should not be construed as limiting the present invention.
[0014] In the description of the present invention, it is necessary to understand that the terms "upper", "lower", "horizontal", "inside", "outside", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.
[0015] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.
[0016] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments.
[0017] Please read Figure 1 The present embodiment provides a fast laser point scanning control device for a large imaging field of view, comprising a laser control module (100) and a dual-axis microcomputer fast scanning mirror (1), a first lens (2), a second lens (3), an X-axis galvanometer scanning mirror (4), a third lens (5), a fourth lens (6), a Y-axis galvanometer scanning mirror (7), a fifth lens (8), a sixth lens (9), a dichroic mirror (10), an objective lens (11) and a detection module (12) arranged in sequence along the propagation direction of an excitation light beam emitted by the laser control module (100), wherein the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) are used for full-width scanning of the entire imaging field of view and switching between different sub-areas within the imaging field of view, and the dual-axis microcomputer fast scanning mirror (1) is used for fast scanning and imaging of a target position, wherein:
[0018] The X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) perform full-width scanning imaging on the entire imaging field of view, and select a sub-region of interest. When the dual-axis microcomputer fast scanning mirror (1) is used to scan and image the sub-region of interest, the X / Y galvanometer scanning mirror is in a stationary state: the excitation light beam emitted by the laser control module is reflected by the dual-axis microcomputer fast scanning mirror (1), then transmitted through the first lens (2) and the second lens (3), and then incident on the X-axis galvanometer scanning mirror (4), and then reflected by the X-axis galvanometer scanning mirror (4), transmitted through the third lens (5) and the fourth lens (6), and then incident on the Y-axis galvanometer scanning mirror (7). The galvanometer scanning mirror (7) is incident on the fifth lens (8) after being reflected by the Y-axis galvanometer scanning mirror (7), and then sequentially focuses on the tissue to be tested through the sixth lens (9), the dichroic mirror (10) and the objective lens (11); the emission light generated by the excitation is then transmitted through the objective lens (11), and then detected by the detection module (12) after being reflected by the dichroic mirror (10), thereby completing the scanning imaging of the sub-area; subsequently, the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) switch between different selected sub-areas, and the dual-axis microcomputer fast scanning mirror (1) scans and images the new sub-area; and the scanning imaging of all sub-areas is completed in a cycle.
[0019] The fluorescent protein mCherry and the calcium ion indicator GCAMP are used to mark the mouse brain nerve cells and the calcium activity of the mouse brain nerve cells respectively, and the X-axis galvanometer scanning mirror and the Y-axis galvanometer scanning mirror are used to fully scan and image the nerve cell structure in the entire imaging field, and the position of the subsequent neural calcium signal imaging sub-area is determined, and the neural calcium signal of the imaging sub-area is quickly scanned and imaged using a dual-axis microcomputer fast scanning mirror. The specific working method is as follows:
[0020] A 1080-nanometer femtosecond laser is used as the excitation light source for two-photon imaging. When the entire imaging field of view is scanned and imaged for the neural cell structure (the MEMS scanning mirror is stationary at this time), the excitation light beam is reflected by the MEMS scanning mirror, and then transmitted through the optical relay composed of the first lens and the second lens; then, the excitation light beam is incident on the X-axis scanning mirror, and after being reflected by the X-axis scanning mirror, it is transmitted through the optical relay composed of the third lens and the fourth lens, and then incident on the Y-axis scanning mirror; the excitation light beam is reflected by the Y-axis scanning mirror and enters the microscopic imaging module (composed of the fifth lens, the sixth lens and the objective lens), and is focused on the mouse brain tissue; the fluorescence generated by the excitation of mCherry is collected by the objective lens, and after being reflected by the dichroic mirror, it is detected by the detection module, completing the scanning imaging of the neural cell structure in the entire imaging field of view. The imaging sub-area for neural calcium activity detection is determined based on the structural image.
[0021] A 920-nanometer femtosecond laser is used as the excitation light source for its two-photon imaging. A MEMS scanning mirror is used to quickly image the neural calcium activity in the imaging sub-area (the X / Y axis scanning mirror is stationary at this time). The excitation beam is reflected by the MEMS scanning mirror, and then transmitted through the optical relay composed of the first lens and the second lens; then, the excitation beam is incident on the X-axis scanning mirror, and after being reflected by the X-axis scanning mirror, it is transmitted through the optical relay composed of the third lens and the fourth lens, and then incident on the Y-axis scanning mirror; the excitation beam is reflected by the Y-axis scanning mirror and enters the microscopic imaging module (composed of the fifth lens, the sixth lens and the objective lens), and is focused in the mouse brain tissue; the fluorescence generated by the excitation GCaMP is collected by the objective lens, and after being reflected by the dichroic mirror, it is detected by the detection module, completing the scanning imaging of the neural calcium activity in the imaging sub-area. Subsequently, the X / Y scanning mirror switches the selected different sub-areas, and uses the MEMS scanning mirror to quickly image the neural calcium activity, and the cycle is repeated.
[0022] It can be understood that the technical features of the above-described embodiments can be arbitrarily combined. In order to make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0023] The above are only preferred embodiments of the present invention, and only specifically describe the technical principles of the present invention. These descriptions are only for explaining the principles of the present invention and cannot be interpreted as limiting the scope of protection of the present invention in any way. Based on the explanations here, any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention, and other specific implementations of the present invention that can be associated with by technicians in this field without creative labor, should be included in the scope of protection of the present invention.
Claims
1. A fast laser point scanning control device for a large imaging field of view, characterized in that: The invention comprises a laser control module (100), and a biaxial microcomputer fast scanning mirror (1), a first lens (2), a second lens (3), an X-axis galvanometer scanning mirror (4), a third lens (5), a fourth lens (6), a Y-axis galvanometer scanning mirror (7), a fifth lens (8), a sixth lens (9), a dichroic mirror (10), an objective lens (11), and a detection module (12) which are sequentially arranged along the propagation direction of an excitation light beam emitted by the laser control module (100), wherein the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) are used for full-width scanning of the entire imaging field of view and switching between different sub-areas in the imaging field of view, and the biaxial microcomputer fast scanning mirror (1) is used for fast scanning and imaging of a target position, wherein: The X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) perform full-width scanning imaging on the entire imaging field of view, and select a sub-region of interest. When the dual-axis microcomputer fast scanning mirror (1) is used to scan and image the sub-region of interest, the X / Y galvanometer scanning mirror is in a stationary state: the excitation light beam emitted by the laser control module is reflected by the dual-axis microcomputer fast scanning mirror (1), then transmitted through the first lens (2) and the second lens (3), and then incident on the X-axis galvanometer scanning mirror (4), and then reflected by the X-axis galvanometer scanning mirror (4), transmitted through the third lens (5) and the fourth lens (6), and then incident on the Y-axis galvanometer The scanning mirror (7) is incident on the fifth lens (8) after being reflected by the Y-axis galvanometer scanning mirror (7) and then sequentially focused on the tissue to be tested through the sixth lens (9), the dichroic mirror (10) and the objective lens (11); the emission light generated by the excitation is then transmitted through the objective lens (11), and then detected by the detection module (12) after being reflected by the dichroic mirror (10), thereby completing the scanning imaging of the sub-area; subsequently, the X-axis galvanometer scanning mirror (4) and the Y-axis galvanometer scanning mirror (7) switch between different selected sub-areas, and the dual-axis microcomputer fast scanning mirror (1) scans and images the new sub-area; and the scanning imaging of all sub-areas is completed in a cycle.
2. The fast laser point scanning control device according to claim 1, characterized in that: The first lens and the second lens constitute an optical relay.
3. The fast laser point scanning control device according to claim 1, characterized in that: The third lens and the fourth lens constitute an optical relay.