Caenorhabditis elegans motion information measuring system and method based on speckle images

Abstract

The invention discloses a system and a method for measuring motion information of caenorhabditis elegans based on speckle images. The system comprises a bearing assembly, a laser assembly is arranged on one side of the bearing assembly, and an imaging assembly is arranged on one side, away from the laser assembly, of the bearing assembly; the laser assembly transmits the bearing assembly in the first direction to provide a light source for the imaging assembly. Or the laser assembly and the imaging assembly are located on the same side of the bearing assembly; the laser assembly irradiates the bearing assembly in the second direction to provide a light source for the imaging assembly. The imaging assembly obtains a plurality of original speckle images of the caenorhabditis elegans and transmits the original speckle images to the processing assembly; according to the measurement system provided by the invention, the original speckle image of the caenorhabditis elegans is directly obtained, analysis is carried out according to the original speckle image, the contrast map is determined, the motion information of the caenorhabditis elegans is determined according to the contrast map, and on the premise that the caenorhabditis elegans does not need to be transferred, the measurement accuracy of the caenorhabditis elegans is improved. And rapid and objective evaluation of nematode motion information is realized.

Description

Technical Field

[0001] This invention relates to the field of motion information technology for Caenorhabditis elegans, and more particularly to a motion information measurement system and method for Caenorhabditis elegans based on speckle images. Background Technology

[0002] Caenorhabditis elegans, due to its well-defined genetic background, short lifespan, and high conservation of molecular pathways with humans, has become an important model organism for studying aging and neurodegenerative diseases such as Alzheimer's disease. In these studies, motor behavior is a key phenotypic indicator for assessing neurological function and overall health. However, current techniques for quantitatively measuring nematode motor ability still have significant limitations in terms of throughput, objectivity, and non-invasiveness, which can be mainly divided into two categories:

[0003] (1) Manual observation and counting methods:

[0004] The throughput is extremely low, and it relies heavily on the operator's experience and judgment, inevitably introducing subjective bias, resulting in poor data repeatability and reliability, making it difficult to meet the needs of modern high-throughput research.

[0005] (2) Automated tracking and analysis system:

[0006] To overcome the shortcomings of manual methods, various automated systems have been developed. However, based on their technological paths, certain inherent bottlenecks still exist: for example, systems based on image analysis and systems based on infrared microbeam detection, but the core principles and application models bring new limitations. Summary of the Invention

[0007] This invention provides a system and method for measuring the movement information of *C. elegans* based on speckle images. The system uses a laser component to vertically transmit light through a carrier component of *C. elegans*, and an imaging component continuously captures multiple raw speckle images. These raw speckle images are then transmitted to a processing component to determine the movement information of *C. elegans*. This method directly acquires raw speckle images without causing physiological disturbances and stress responses by transferring the *C. elegans* to a liquid environment. A contrast map is then determined from these raw speckle images to obtain the movement information of *C. elegans*, enabling continuous, non-invasive, long-term behavioral monitoring of *C. elegans* throughout its entire life cycle.

[0008] According to a first aspect of the present invention, a motion information measurement system for Caenorhabditis elegans based on speckle images is provided, comprising: a laser component, a carrier component, an imaging component, and a processing component;

[0009] The carrier component is used to carry the Caenorhabditis elegans worm to be tested;

[0010] The laser component is disposed on one side of the support component, and the imaging component is disposed on the side of the support component opposite to the laser component; the output end of the imaging component is connected to the input end of the processing component.

[0011] The laser component is used to transmit the carrier component along a first direction to provide a light source for the imaging component;

[0012] Alternatively, the laser component and the imaging component are located on the same side of the carrier component; the output end of the imaging component is connected to the input end of the processing component; the laser component is used to irradiate the carrier component along a second direction to provide a light source for the imaging component;

[0013] The imaging component acquires multiple raw speckle images of the Caenorhabditis elegans and transmits them to the processing component; the processing component determines the motion information of the Caenorhabditis elegans based on the raw speckle images; wherein, the first direction is perpendicular to the bearing surface of the bearing component, and the second direction intersects with the first direction.

[0014] Optionally, the laser assembly includes a laser, a first lens, and a plurality of scattering sheets;

[0015] The laser is used to emit a laser beam, which is incident on the first lens and, after passing through a plurality of the scattering sheets, is incident on the carrier component along the first direction or the second direction.

[0016] Optionally, the first lens may include a plano-concave lens.

[0017] Optionally, the imaging component includes a complementary metal-oxide-semiconductor camera or a charge-coupled device camera.

[0018] Optionally, the carrier component includes a solid culture medium for Caenorhabditis elegans.

[0019] Optionally, the aperture coefficient of the imaging component includes 2.8.

[0020] According to a second aspect of the present invention, a method for measuring the motion information of *Caenorhabditis elegans* based on speckle images is provided, applicable to any of the *Caenorhabditis elegans* motion information measurement systems based on speckle images described in the first aspect of the present invention, the method comprising:

[0021] Obtain raw speckle images of Caenorhabditis elegans;

[0022] The speckle image sequence is determined based on the original speckle image and the static background intensity map;

[0023] The contrast map of the *Caenorhabditis elegans* was determined based on the speckle image sequence;

[0024] The movement information of the *Caenorhabditis elegans* was determined based on the contrast map.

[0025] Optionally, determining the speckle image sequence based on the original speckle image and the static background intensity map includes:

[0026] Divide the pixels of each frame of the original speckle image in the original speckle image by the static background light intensity map to obtain the speckle image sequence.

[0027] Optionally, determining the contrast map of *Caenorhabditis elegans* based on the speckle image sequence includes:

[0028] The standard deviation of light intensity of each pixel in the time dimension is determined based on each pixel in the speckle image sequence;

[0029] The contrast map of the *Caenorhabditis elegans* is determined based on the light intensity standard deviation and the average light intensity of the pixels.

[0030] Optionally, determining the contrast map of *Caenorhabditis elegans* based on the light intensity standard deviation and the average light intensity of the pixels includes:

[0031] The contrast map is obtained by dividing the standard deviation of light intensity of each pixel in the speckle image sequence by the average light intensity of each pixel in the speckle image sequence.

[0032] This invention discloses a system and method for measuring the motion information of *C. elegans* based on speckle images, comprising: a laser component, a carrier component, an imaging component, and a processing component; the carrier component is used to carry the *C. elegans* to be measured; in a first direction, a laser component is disposed on one side of the carrier component, and an imaging component is disposed on the side of the carrier component opposite to the laser component; the output end of the imaging component is connected to the input end of the processing component; the laser component is used to transmit light from the carrier component along the first direction to provide a light source for the imaging component; the imaging component acquires multiple original speckle images of *C. elegans* and transmits them to the processing component; the processing component determines the motion information of *C. elegans* based on the original speckle images; wherein, the first direction is the direction perpendicular to the carrier surface of the carrier component. The present invention provides a Caenorhabditis elegans motion information measurement system based on speckle images. This system uses a laser component to vertically transmit light through a carrier component of the nematode, and an imaging component to continuously capture multiple raw speckle images. These raw speckle images are then transmitted to a processing component to determine the motion information of the nematode. This system directly acquires raw speckle images without causing physiological disturbances and stress responses by transferring the nematode to a liquid environment. A contrast map is then determined from these raw speckle images to obtain the motion information of the nematode, enabling continuous, non-invasive, long-term behavioral monitoring of the nematode throughout its entire life cycle.

[0033] It should be understood that the description in this section is not intended to identify key or essential features of the embodiments of the present invention, nor is it intended to limit the scope of the invention. Other features of the invention will become readily apparent from the following description. Attached Figure Description

[0034] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0035] Figure 1 This is a structural diagram of a Caenorhabditis elegans motion information measurement system based on speckle images provided in an embodiment of the present invention;

[0036] Figure 2 This is a structural diagram of a Caenorhabditis elegans motion information measurement system based on speckle images provided in an embodiment of the present invention;

[0037] Figure 3 This is a flowchart of a method for measuring the motion information of Caenorhabditis elegans based on speckle images, provided by an embodiment of the present invention.

[0038] Figure 4This is a schematic diagram of the spatial contrast principle in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention.

[0039] Figure 5 This is a schematic diagram of the temporal contrast in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention.

[0040] Figure 6 This is a schematic diagram of the spatiotemporal mixing contrast in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention.

[0041] Figure 7 This is a motion image of Caenorhabditis elegans provided in an embodiment of the present invention, based on a speckle image-based motion information measurement system for Caenorhabditis elegans.

[0042] Figure 8 This is a motion image of Caenorhabditis elegans from another speckle image-based motion information measurement system provided in this embodiment of the invention.

[0043] Figure 9 This is a motion image of Caenorhabditis elegans from another speckle image-based motion information measurement system provided in this embodiment of the invention.

[0044] Figure 10 This is a motion image of Caenorhabditis elegans from another speckle image-based motion information measurement system provided in this embodiment of the invention.

[0045] Figure 11 This is a flowchart of another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention;

[0046] Figure 12 This is the original speckle image in another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention;

[0047] Figure 13 This is a static background light intensity map in another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention;

[0048] Figure 14 Another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention includes a speckle image sequence.

[0049] Figure 15 This is a flowchart of another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention;

[0050] Figure 16This is a flowchart of another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention. Detailed Implementation

[0051] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0052] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0053] It should be understood that the various forms of processes shown above can be used, with steps reordered, added, or deleted. For example, the steps described in this invention can be executed in parallel, sequentially, or in different orders, as long as the desired result of the technical solution of this invention can be achieved, and this is not limited herein.

[0054] In existing technologies, numerous automated tracking and analysis systems have been developed to overcome the unreliability of manual observation:

[0055] One type is image-based systems. While these systems increase throughput, their data processing often involves computationally intensive steps, such as image segmentation, skeleton extraction, and multi-parameter feature calculation. The analysis process is complex and time-consuming, requiring rigorous professional training for users and typically relying on expensive dedicated imaging hardware or precision sensors. More fundamentally, many systems, for ease of observation, often require transferring nematodes from their normal growth medium (Nematode Growth Medium, NGM) to a liquid environment (such as M9 buffer) or specific microplates. This physical transfer itself is a strong stressor, interfering with the nematodes' true physiological state and thus affecting the accuracy of behavioral data.

[0056] Another type is the infrared microbeam detection system. While this approach doesn't require complex image processing, its core principles and application modes introduce new limitations. First, these instruments primarily generate activity signals by detecting the occlusion or scattering effects of biological movement on the infrared beam, converting these signals into abstract parameters such as movement speed and activity index. This method is essentially an indirect measurement, unable to provide intuitive images containing the nematode's true morphology, precise coordinates, and posture information, making it difficult for researchers to verify and conduct in-depth visual analysis of the nematode's specific behavioral state. Furthermore, to ensure baseline stability and accuracy of the detection signal, some models of this series of instruments and most applications still require measurements to be performed in liquid media. This not only differs from the natural behavior of nematodes on solid culture media but also introduces stress interference from the transfer operation. Second, these systems typically require transferring the tested nematodes to relatively confined spaces such as multi-well plates or small culture dishes for detection. This detection environment not only limits the population size for a single observation but also makes it difficult to assess the collective behavior of nematodes under more open, near-conventional culture conditions.

[0057] In summary, the core drawbacks of both traditional manual techniques and mainstream automated systems lie in the invasiveness of the operation and the resulting discontinuity in monitoring. For example, many methods, for ease of observation, typically require transferring nematodes from their normal growth medium (such as NGM) to a liquid environment or specific microplates. This physical transfer itself is a strong stressor, interfering with the nematodes' true physiological state. Since the transfer process is often irreversible, existing techniques struggle to conduct long-term, continuous longitudinal observations of the same population, which is crucial for studying the progression of chronic diseases or assessing the long-term effects of drugs. Therefore, there is an urgent need in this field for a novel, non-invasive, in-situ method for direct detection of nematodes on solid culture media.

[0058] Therefore, this invention provides a motion information measurement system for Caenorhabditis elegans based on speckle images. Figure 1 This is a structural diagram of a Caenorhabditis elegans motion information measurement system based on speckle images provided in an embodiment of the present invention; Figure 2 This is a structural diagram of another motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention, with reference to... Figure 1 and Figure 2 The motion information measurement system of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention includes: a laser component 1, a carrier component 2, an imaging component 3, and a processing component 4; the carrier component 2 is used to carry the Caenorhabditis elegans 5 to be tested;

[0059] A laser component 1 is disposed on one side of the support component 2, and an imaging component 3 (i.e., Figure 1 (as shown); the output end of the imaging component 3 is connected to the input end of the processing component 4; the laser component 1 is used to transmit the load-bearing component 2 along the first direction x to provide a light source for the imaging component 3;

[0060] Alternatively, the laser assembly 1 and the imaging assembly 3 are located on the same side of the carrier assembly 2 (i.e., Figure 2 (as shown); the output end of the imaging component 3 is connected to the input end of the processing component 4; the laser component 1 is used to irradiate the carrier component 2 along the second direction y to provide a light source for the imaging component 3;

[0061] Imaging component 3 acquires multiple raw speckle images 6 of Caenorhabditis elegans 5 and transmits them to processing component 4; processing component 4 determines the motion information of Caenorhabditis elegans 5 based on the raw speckle images 6; wherein, the first direction x is perpendicular to the bearing surface of bearing component 2, and the second direction y intersects the first direction x.

[0062] Specifically, the measurement system provided in this embodiment of the invention includes: a laser component 1, a carrier component 2, an imaging component 3, and a processing component 4;

[0063] The carrier component 2 is used to carry the Caenorhabditis elegans 5 to be tested. The carrier component 2 can be the Caenorhabditis elegans growth medium (NGM). NGM medium is a solid culture medium specially prepared for culturing Caenorhabditis elegans. Its main components include sodium chloride, peptone, cholesterol and agar, in order to carry and culture the Caenorhabditis elegans 5 to be tested.

[0064] like Figure 1 As shown, in the first direction x, which is perpendicular to the bearing surface of the bearing component 2, a laser component 1 is provided on one side of the bearing component 2, a laser component 1 is provided on one side of the bearing component 2 (i.e., below the bearing component 2), and an imaging component 3 is provided on the side of the bearing component 2 away from the laser component 1 (i.e. above the bearing component 2); wherein, the laser component 1, the bearing component 2 and the imaging component 3 are all distributed along the first direction x;

[0065] The output of imaging component 3 is connected to the input of processing component 4;

[0066] The laser assembly 1 is used to transmit light from the carrier assembly 2 along the first direction x to provide a light source for the imaging assembly 3; that is, the laser assembly 1 is perpendicular to the carrier assembly 2 to illuminate the carrier assembly 2. The measurement system also includes a current driver. Figure 1(Not shown in the image) The current driver provides energy for the laser beam emitted by the laser component 1; the imaging component 3 captures a series (e.g., 50 frames) of multiple consecutive raw speckle images 6 above the supporting component 2, and transmits them to the input of the processing component 4 through the output of the imaging component 3. The processing component 4 can be a computer. The processing component 4 determines the contrast map of Caenorhabditis elegans 5 based on the multiple raw speckle images 6 captured by the imaging component 3, and determines the motion information of Caenorhabditis elegans 5 based on the contrast map; the processing component 4 can also control the parameter information of the imaging component 3 (such as exposure time and frame rate).

[0067] like Figure 2 As shown, the laser component 1 and the imaging component 3 are located on the same side of the supporting component 2 (i.e., Figure 2 The system also includes a current driver (above the center) to provide illumination for the carrier component 2. Figure 2 (Not shown in the image) The current driver provides energy for the laser beam emitted by the laser component 1; the imaging component 3 captures a series (e.g., 50 frames) of multiple consecutive raw speckle images 6 above the supporting component 2, and transmits them to the input of the processing component 4 through the output of the imaging component 3. The processing component 4 can be a computer. The processing component 4 determines the contrast map of Caenorhabditis elegans 5 based on the multiple raw speckle images 6 captured by the imaging component 3, and determines the motion information of Caenorhabditis elegans 5 based on the contrast map; the processing component 4 can also control the parameter information of the imaging component 3 (such as exposure time and frame rate); the measurement system provided in this embodiment of the invention needs to be placed in a closed, light-shielding environment to isolate the interference of ambient light and ensure the stability and accuracy of the measurement.

[0068] The measurement system provided in this embodiment of the invention continuously captures multiple raw speckle images through an imaging component and transmits the raw speckle images to a processing component to determine the movement information of *C. elegans*. This allows for the direct acquisition of raw speckle images without causing physiological disturbances and stress responses by transferring *C. elegans* to a liquid environment. A contrast map is then determined from the raw speckle images to obtain the movement information of *C. elegans*, enabling continuous, non-invasive, long-term behavioral monitoring of *C. elegans* throughout its entire life cycle.

[0069] Optional, continue to refer to Figure 1 and Figure 2 The laser component 1 in the motion information measurement system of Caenorhabditis elegans provided in this embodiment of the invention includes a laser 11, a first lens 12 and multiple scattering sheets 13;

[0070] The laser 11 is used to emit a laser beam. The laser beam is incident on the first lens 12 and, after passing through multiple scatterers 13, is incident on the carrier component 2 along the first direction X or the second direction Y.

[0071] Specifically, the laser assembly 1 includes a laser 11, a first lens 12, and multiple scattering sheets 13;

[0072] Along the side of the support component 2 that faces away from the imaging component 3, i.e. Figure 1 In the middle, multiple scattering plates 13 are distributed sequentially below the supporting component 2. Figure 1 and Figure 2 The example shows three diffusers 13), a first lens 12, and a laser 11;

[0073] The laser 11 can be a laser diode, a solid-state laser, etc. The laser 11 is used to emit a laser beam. The laser beam is incident on the first lens 12. After the laser beam is transmitted through the first lens 12, it diverges and expands before being incident on the scatterer 13. After the diverged and expanded laser beam passes through the scatterer 13, it becomes a uniform and soft laser beam and is then incident on the bottom of the support component 2 to provide a light source for the support component 2, so as to illuminate the support component 2 and the Caenorhabditis elegans 5, so that the imaging component 3 can better capture the original speckle image 6.

[0074] Optionally, the first lens 12 in the motion information measurement system of Caenorhabditis elegans provided in this embodiment of the invention includes a plano-concave lens.

[0075] Specifically, a plano-concave lens has a flat surface on one side and an inwardly concave curved surface on the other. When a laser beam parallel to the principal axis passes through the lens, the concave surface causes the light to bend in all directions. Because the lens is thinner in the middle than at the edges, the light is delayed less in the middle, causing the emitted light rays to not converge at a single point, thus amplifying the laser beam.

[0076] Optionally, the imaging component 3 in the motion information measurement system for Caenorhabditis elegans provided in this embodiment of the invention includes a complementary metal-oxide-semiconductor camera or a charge-coupled device camera.

[0077] Specifically, the imaging component 3 can be a complementary metal-oxide-semiconductor (CMOS) camera or a charge-coupled device (CCD) camera to continuously acquire multiple raw speckle images of Caenorhabditis elegans 5.

[0078] Optionally, the carrier component 2 includes a solid culture medium for Caenorhabditis elegans.

[0079] Optionally, the wavelength of the laser beam is 780nm.

[0080] Optionally, the aperture of imaging component 3 includes 2.8.

[0081] Based on the same inventive concept, embodiments of the present invention provide a method for measuring the motion information of *Caenorhabditis elegans* based on speckle images, applicable to any of the above-described embodiments of the *Caenorhabditis elegans* motion information measurement system based on speckle images. Figure 3 This is a flowchart of a method for measuring the motion information of *Caenorhabditis elegans* based on speckle images, provided in an embodiment of the present invention. (Refer to...) Figures 1-3 The present invention provides a method for measuring the motion information of Caenorhabditis elegans based on speckle images, comprising:

[0082] S101. Obtain the original speckle image of Caenorhabditis elegans.

[0083] Specifically, before step S101, the Caenorhabditis elegans 5 to be tested is placed on its conventional growth medium NGM (carrier component 2), and laser component 1 is used to vertically transmit light to carrier component 2 from below.

[0084] Imaging component 3 captures a series (e.g., 50 frames) of consecutive raw speckle images 6 above carrier component 2 and transmits them to processing component 4.

[0085] S102. Determine the speckle image sequence based on the original speckle image and the static background light intensity map.

[0086] Specifically, in order to eliminate the influence of uneven lighting, a speckle image sequence (the corrected ideal speckle image sequence) is determined by the original speckle image 6 obtained in step S101 above and the static background light intensity map.

[0087] Among them, the static background light intensity map can be obtained by averaging a large number of speckle images of static Caenorhabditis elegans (such as blank culture medium) without any active organisms.

[0088] S103. Determine the contrast image of Caenorhabditis elegans based on the speckle image sequence.

[0089] Specifically, this invention uses the Temporal Laser Speckle Contrast algorithm as its theoretical basis. It determines the contrast map of *Caenorhabditis elegans* through a sequence of speckle images. Each pixel value in this contrast map directly reflects the intensity of motion at its corresponding spatial location. The faster the *C. elegans* moves, the more drastic the speckle changes, the stronger the time-averaging effect, and the higher the contrast map K. t The lower the value, the better.

[0090] Figure 4 This is a schematic diagram of the spatial contrast principle in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention. Figure 5This is a schematic diagram of the temporal contrast in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention. Figure 6 This is a schematic diagram of the spatiotemporal mixing contrast in a motion information measurement system for Caenorhabditis elegans based on speckle images provided in an embodiment of the present invention. (Refer to...) Figures 4-6 , Figure 4 Spatial contrast (K) s Schematic diagram; Figure 5 It is time contrast (K) t Schematic diagram; Figure 6 It is a spatiotemporal mixing contrast (K st / K ts Schematic diagram.

[0091] S104. Determine the movement information of Caenorhabditis elegans based on the contrast diagram.

[0092] Specifically, in the generated contrast K t In the figure, the distinction between the target and the background depends on the difference in their dynamic characteristics. The static NGM (C. elegans solid culture medium) background, due to its near-transparency and the absence of moving scattering particles within it, produces almost no dynamic speckle patterns under coherent light illumination, resulting in an extremely low time Kt value, even close to zero.

[0093] Conversely, the continuous peristalsis and internal physiological activities of active *Caenorhabditis elegans* cause dynamic scattering of transmitted light, resulting in a highly fluctuating temporal speckle signal. This makes the Kt value of the area where the active nematode is located significantly higher than that of the static background. Therefore, in the final generated Kt map, the active nematode appears as a clear, high-contrast outline, while the static background is a low-contrast area, creating a sharp contrast that makes them easily identifiable.

[0094] At this point, only simple image processing methods are needed, such as setting a global intensity threshold to filter out background artifacts with low Kt values, or by minimizing manual interaction (such as ROI selection), to accurately and conveniently separate the nematode region.

[0095] The average Kt value of all pixels within a segmented individual nematode region is calculated to obtain a single, quantifiable motion index, which characterizes the overall mobility of the Caenorhabditis elegans.

[0096] Figure 7 This is a motion image of Caenorhabditis elegans provided in an embodiment of the present invention, based on a speckle image-based motion information measurement system for Caenorhabditis elegans. Figure 8 This is a motion image of Caenorhabditis elegans from another speckle image-based motion information measurement system provided in this embodiment of the invention. Figure 9This is a motion image of Caenorhabditis elegans from another speckle image-based motion information measurement system provided in this embodiment of the invention. Figure 10 This is a motion image of *Caenorhabditis elegans* from another *Caenorhabditis elegans* motion information measurement system based on speckle images provided in this embodiment of the invention. (Refer to...) Figures 7-10 The diagrams illustrate the image segmentation and quantification process based on the region of interest (ROI); the process includes:

[0097] Figure 7 : Use bounding boxes to manually locate target individuals;

[0098] Figure 8 Extract precise coordinates from the selected area;

[0099] Figure 9 Define the cropped ROI around the target;

[0100] Figure 10 Apply a threshold segmentation algorithm within the ROI to obtain the final segmented objects.

[0101] This invention provides a method for measuring the motion information of Caenorhabditis elegans based on speckle images;

[0102] 1. Experimental setup:

[0103] System parameters: The system for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention is used. The wavelength of the laser beam is set to 780nm, the exposure time of the imaging component is set to 0.3ms, and the lens aperture parameter is 2.8.

[0104] 2. Experimental Groups: Three experimental groups were set up: (1) Aβ-induced untreated group (disease model group); (2) Aβ-induced drug-treated group (drug efficacy group); (3) Non-induced control group (normal aging group). Each group had 3 biological replicates, with approximately 15 Caenorhabditis elegans per replicate.

[0105] 2. Experimental Procedure:

[0106] 1) All synchronized L1 stage larvae in all groups were first cultured on NGM medium at 15°C, at which Aβ gene expression was suppressed.

[0107] 2) Starting from time t=0, data is collected from each group every 12 hours using the Caenorhabditis elegans motion information measurement system based on speckle images provided in this embodiment of the invention.

[0108] 3) At 36 hours (L2 / L3 development stage), the culture dishes of the disease model group and the drug efficacy group were transferred to 25°C to induce the expression of Aβ protein, thereby inducing progressive paralysis.

[0109] 4) Starting from the 60th hour (24 hours after induction), as Caenorhabditis elegans begins to show signs of paralysis, the observation interval is shortened to 6 hours to accurately capture key points of phenotypic changes.

[0110] 5) During each data acquisition, capture 50 consecutive frames of raw speckle images for subsequent analysis.

[0111] 3. Data Analysis and Results

[0112] For each acquired speckle image sequence, the method provided in this embodiment of the invention is applied: K is calculated. t The graph was finally segmented by ROI and the average K value for each nematode was calculated. t value.

[0113] The results show that:

[0114] Control group: its average K t The value showed a slow, linear increase over time (0-84 hours), which is consistent with the known pattern of natural decline in locomotor ability caused by physiological aging in Caenorhabditis elegans, verifying the effectiveness of this method as an aging biomarker.

[0115] Disease model group: After 36 hours of temperature upregulation, its K t The values ​​initially rose briefly (related to increased temperature and accelerated development), but then dropped sharply after 60 hours, reaching a plateau at approximately 66 hours, indicating that the nematode population had generally entered a state of complete paralysis. This precisely quantifies the severe loss of motor function caused by Aβ toxicity.

[0116] Drug efficacy group: Compared with the disease model group, the nematode K in this group... t The rate of decrease in its K value slowed significantly, and its K value decreased significantly. t The time it took for the value to reach its trough (approximately 72 hours) was significantly later than in the disease model group. This quantitatively demonstrates that the compound used can effectively delay the Aβ-induced paralysis process and exert a neuroprotective effect.

[0117] Furthermore, during the transition period of paralysis (e.g., 66 hours), the K values ​​in the drug group and the untreated group were significantly different. t The data exhibits a non-normal distribution, directly reflecting the phenotypic heterogeneity within the population (partial paralysis, partial activity), providing richer biological information for assessing disease progression and drug efficacy than a single average.

[0118] The method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this invention can be used for:

[0119] Aging biology research: By conducting long-term, continuous, non-invasive monitoring of the same nematode population, the natural decline in motor ability caused by physiological aging throughout its life cycle can be precisely quantified, providing objective and dynamic evaluation indicators for assessing interventions to delay aging (such as drugs and gene editing).

[0120] ● Modeling and research of neurodegenerative diseases: This technology can be widely applied to nematode models of various neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease, Huntington's disease, etc.) to dynamically track the disease progression, non-invasively assess motor dysfunction caused by protein toxicity or neuronal damage, and analyze their potential pathophysiological mechanisms.

[0121] ● Large-scale drug screening and gene function validation: With its high throughput, automation potential and objective quantification, this platform can efficiently screen compound libraries or perform genetic screening to quickly discover potential therapeutic drugs or key regulatory genes that can improve or delay specific pathological phenotypes (such as Aβ-induced paralysis), greatly accelerating the process of new drug development and functional genomics research.

[0122] To further clarify the technical solution, effects, and advantages of the present invention, this paper will illustrate with a specific application example: using the system and method described in this invention, high-resolution dynamic monitoring and quantitative evaluation were performed on the paralysis process induced by Aβ protein toxicity in an Alzheimer's disease Caenorhabditis elegans model, as well as the intervention effect of a specific drug.

[0123] Based on the above embodiments, the present invention further refines the determination of the speckle image sequence based on the original speckle image and the static background intensity map. Figure 11 This is a flowchart of another method for measuring the motion information of *Caenorhabditis elegans* based on speckle images provided in this embodiment of the invention. (Refer to...) Figure 11 The method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention includes:

[0124] S201. Obtain the original speckle image of Caenorhabditis elegans.

[0125] S202. Determine the speckle image sequence based on the original speckle image and the static background light intensity map.

[0126] S203. Divide the pixels of each frame of the original speckle image in the original speckle image by the static background light intensity map to obtain the speckle image sequence.

[0127] Specifically, Figure 12This is the original speckle image in another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention; Figure 13 This is a static background light intensity map in another method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention; Figure 14 Another method for measuring the motion information of *Caenorhabditis elegans* based on speckle images provided in this embodiment of the invention includes a speckle image sequence diagram; see reference. Figures 12-14 Each frame of the original speckle image is divided by a pre-acquired static background light intensity map to obtain a speckle image sequence of Caenorhabditis elegans.

[0128] S204. Determine the contrast image of Caenorhabditis elegans based on the speckle image sequence.

[0129] S205. Determine the movement information of Caenorhabditis elegans based on the contrast diagram.

[0130] Based on the above embodiments, this invention further refines the determination of the contrast map of *Caenorhabditis elegans* from speckle image sequences. Figure 15 This is a flowchart of another method for measuring the motion information of *Caenorhabditis elegans* based on speckle images provided in this embodiment of the invention. (Refer to...) Figure 15 The method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention includes:

[0131] S301. Obtain the original speckle image of Caenorhabditis elegans.

[0132] S302. Determine the speckle image sequence based on the original speckle image and the static background light intensity map.

[0133] S303. Determine the standard deviation of light intensity of each pixel in the time dimension based on each pixel in the speckle image sequence.

[0134] Specifically, based on the temporal speckle contrast method, the standard deviation of light intensity in the time dimension is calculated for each pixel in the speckle image sequence.

[0135] S304. Determine the contrast map of Caenorhabditis elegans based on the standard deviation of light intensity and the average light intensity of the pixels.

[0136] Specifically, based on the temporal speckle contrast method, the average light intensity of each pixel in the speckle image sequence is calculated in the time dimension, and the contrast map of Caenorhabditis elegans is determined according to the standard deviation of light intensity and the average light intensity.

[0137] S305. Determine the movement information of Caenorhabditis elegans based on the contrast diagram.

[0138] Based on the above embodiments, this invention further refines the determination of the contrast map of *Caenorhabditis elegans* based on the standard deviation of light intensity and the average light intensity of pixels. Figure 16 This is a flowchart of another method for measuring the motion information of *Caenorhabditis elegans* based on speckle images provided in this embodiment of the invention. (Refer to...) Figure 16 The method for measuring the motion information of Caenorhabditis elegans based on speckle images provided in this embodiment of the invention includes:

[0139] S401. Obtain the original speckle image of Caenorhabditis elegans.

[0140] S402. Determine the speckle image sequence based on the original speckle image and the static background light intensity map.

[0141] S403. Determine the standard deviation of light intensity of each pixel in the time dimension based on each pixel in the speckle image sequence.

[0142] S404. Determine the contrast map of Caenorhabditis elegans based on the standard deviation of light intensity and the average light intensity of the pixels.

[0143] S405. Divide the standard deviation of light intensity of each pixel in the speckle image sequence by the average light intensity of each pixel in the speckle image sequence to obtain a contrast map.

[0144] Specifically, the standard deviation of light intensity of each pixel in the speckle image sequence obtained in step S404 is divided by the average light intensity of each pixel in the speckle image sequence to obtain a two-dimensional contrast K. t The image shows that each pixel value directly reflects the intensity of motion at the corresponding spatial location—the faster the motion, the more dramatic the speckle changes, and the stronger the time averaging effect. t The lower the value.

[0145] The specific embodiments described above do not constitute a limitation on the scope of protection of this invention. Those skilled in the art should understand that various modifications, combinations, sub-combinations, and substitutions can be made according to design requirements and other factors. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this invention should be included within the scope of protection of this invention.

Claims

1. A system for measuring information of Caenorhabditis elegans movement based on speckle images, characterized by, The system comprises a laser assembly, a bearing assembly, an imaging assembly and a processing assembly; The bearing assembly is used for bearing the Caenorhabditis elegans to be measured; The bearing assembly is provided with the laser assembly on one side, and the imaging assembly is provided on the side of the bearing assembly away from the laser assembly; the output end of the imaging assembly is connected with the input end of the processing assembly; The laser assembly is used for transmitting the bearing assembly along a first direction to provide a light source for the imaging assembly; Alternatively, the laser assembly and the imaging assembly are located on the same side of the bearing assembly; the output end of the imaging assembly is connected with the input end of the processing assembly; and the laser assembly is used for irradiating the bearing assembly along a second direction to provide a light source for the imaging assembly. The imaging assembly acquires original speckle images of a plurality of Caenorhabditis elegans and transmits the original speckle images to the processing assembly; the processing assembly determines motion information of the Caenorhabditis elegans according to the original speckle images; wherein the first direction is a direction perpendicular to a bearing surface of the bearing assembly, and the second direction intersects the first direction.

2. The speckle image-based C. elegans locomotion information measurement system according to claim 1, characterized by, The laser assembly comprises a laser, a first lens and a plurality of scattering pieces. The laser is used for emitting a laser beam; the laser beam is incident to the first lens and then is incident to the bearing assembly along the first direction or the second direction after passing through the plurality of scattering pieces.

3. The speckle image-based C. elegans locomotion information measurement system according to claim 2, characterized by, The first lens comprises a plano-concave lens.

4. The speckle image-based C. elegans locomotion information measurement system according to claim 1, characterized by, The imaging assembly comprises a complementary metal-oxide-semiconductor camera or a charge-coupled device camera.

5. The speckle pattern image based C. elegans locomotion information measurement system according to claim 1, characterized by, The bearing assembly comprises a Caenorhabditis elegans solid culture medium.

6. The speckle pattern image based C. elegans locomotion information measurement system according to claim 1, wherein An aperture coefficient of the imaging assembly comprises 2.

8.

7. A method of measuring information on movement of Caenorhabditis elegans based on a speckle image, characterized by The method is suitable for the system of any one of claims 1-6. Acquiring original speckle images of Caenorhabditis elegans; Determining a speckle image sequence according to the original speckle images and a static background light intensity map; Determining a contrast map of the Caenorhabditis elegans according to the speckle image sequence; Determining motion information of the Caenorhabditis elegans according to the contrast map.

8. The speckle image-based C. elegans locomotion information measuring method according to claim 7, characterized by, The method of determining the speckle image sequence according to the original speckle images and the static background light intensity map comprises: Dividing each pixel point of each frame of the original speckle images by the static background light intensity map to obtain the speckle image sequence.

9. The speckle image-based C. elegans locomotion information measuring method according to claim 7, characterized by, The method of determining the contrast map of the Caenorhabditis elegans according to the speckle image sequence comprises: Determining a light intensity standard deviation of each pixel point of the speckle image sequence in a time dimension; Determining the contrast map of the Caenorhabditis elegans according to the light intensity standard deviation and an average light intensity of the pixel point.

10. The method of claim 9, wherein the method is a method of measuring Caenorhabditis elegans locomotion information based on a speckle image. The method of determining the contrast map of the Caenorhabditis elegans according to the light intensity standard deviation and the average light intensity of the pixel point comprises: Dividing the light intensity standard deviation of each pixel point of the speckle image sequence by the average light intensity of each pixel point of the speckle image sequence to obtain the contrast map.

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