Single-detector single-scan phase-multiplexed multicolor imaging method and apparatus

Abstract

This application belongs to the field of multicolor imaging technology, specifically disclosing a single-detector, single-scan phase-multiplexed multicolor imaging method and apparatus. The method includes: projecting a line spot onto the surface of a fluorescent sample to be imaged using a laser; using a detector to scan the surface of the fluorescent sample in a strip-scan manner to obtain multi-layer fluorescence signals; during the strip-scanning process, determining the variation period based on the single exposure time of the detector, adjusting the intensity of the laser to vary sinusoidally with the variation period, applying a time delay of a preset wavelength to the intensity variation waveform of the laser to generate a phase with a wavelength identifier; and processing the multi-layer fluorescence signals to obtain the multicolor imaging result. This method can significantly reduce the time consumption of multicolor imaging.

Description

Technical Field

[0001] This application belongs to the field of multicolor imaging technology, and more specifically, relates to a single detector single scan phase multiplexing multicolor imaging method and apparatus. Background Technology

[0002] Multicolor fluorescence microscopy analyzes the distribution of different substances by acquiring the spectral information of each sampling point in the sample. Existing technologies usually use multiple scans to achieve multicolor imaging, including the following two types: 1) Snapshot imaging: In a single exposure time, two-dimensional images at different wavelengths are acquired simultaneously through multiple area array detectors; 2) Alternating imaging: Filter groups are selected according to the color channels, and after scanning one color, another color channel is switched, and imaging is carried out in sequence until all colors have been traversed.

[0003] Although both methods have their advantages, they also have significant drawbacks: 1) Snapshot imaging: When the imaging range exceeds the size of a single field of view and mosaic scanning imaging is required, switching between different mosaic imaging methods and the movement and stabilization of the sample takes a certain amount of time, which reduces the overall imaging efficiency; 2) Alternating imaging: Different colors require switching between different filter groups, and the imaging time increases exponentially with the increase in the number of color channels. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this application is to provide a single detector single scan phase multiplexing multicolor imaging method and apparatus, which aims to solve the problems of high-quality multicolor imaging range being limited by imaging throughput and low imaging efficiency caused by multiple scans in the prior art.

[0005] To achieve the above objectives, in a first aspect, this application provides a single-detector single-scan phase-multiplexed multicolor imaging method, comprising:

[0006] A laser is used to project a line spot onto the surface of the fluorescent sample to be imaged;

[0007] The detector is used to scan the surface of the fluorescent sample to be imaged in a strip scanning manner to obtain multi-layer fluorescence signals. During the strip scanning process, the change period is determined based on the single exposure time of the detector. The intensity of the laser is adjusted to vary sinusoidally according to the change period. A time delay of a preset wavelength is applied to the intensity change waveform of the laser to generate a phase with wavelength marking.

[0008] The multilayer fluorescence signals are processed to obtain multicolor imaging results.

[0009] This application utilizes a detector to perform a single scan of the fluorescent sample surface in a strip scanning manner, while simultaneously adjusting the laser intensity sinusoidally based on the detector's single exposure time. A phase delay is applied to the laser based on its wavelength, resulting in simultaneous temporal and spatial modulation during sample scanning. This allows for the acquisition of data required for phase-multiplexed multicolor imaging in a single scan, increasing system imaging throughput and reducing the time consumption of multicolor imaging methods. Furthermore, operation is performed on the original image from online confocal imaging, preserving optical tomography capabilities and enabling combination with the line scanning method, which boasts the highest scanning throughput, making it highly suitable for large-scale sample imaging.

[0010] According to the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application, the detector includes multiple sub-detectors. During the process of using the detector to scan the surface of the fluorescent sample to be imaged in a strip scanning manner, the multiple sub-detectors in the detector record the fluorescence signals of different sample points under the same light intensity at the same time, and finally record the fluorescence signals of different sample points under different light intensity.

[0011] This application uses multiple sub-detectors to simultaneously record the sample signal within the illumination range, enabling a single camera to obtain all the data of the demodulated monochrome image in a single scan, without additional pauses or the time consumed by multiple scans.

[0012] According to the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application, the step of processing the multilayer fluorescence signal to obtain the multicolor imaging result includes:

[0013] The multilayer fluorescence signal is processed to obtain an intensity map of all fluorescence channels mixed together and a phase map that can be used to distinguish the fluorescence channels;

[0014] Based on the intensity map and the phase map, the fluorescence channel is split to obtain multicolor imaging results.

[0015] According to the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application, the processing of the multilayer fluorescence signal to obtain an intensity map of all fluorescence channels mixed and a phase map that can be used to distinguish fluorescence channels includes:

[0016] The multi-layer fluorescence signals are rearranged according to the time periodic pattern to generate four images taken within one period.

[0017] Based on the four images, the intensity map of all fluorescence channels and the phase map that can be used to distinguish the fluorescence channels are obtained using the four-step phase-shifting method.

[0018] According to the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application, the step of splitting the fluorescence channel based on the intensity map and the phase map to obtain the multicolor imaging result includes:

[0019] The intensity map is binarized, and the obtained mask is multiplied with the phase map to obtain the aliasing intensity map;

[0020] The multiplexed channel is split based on the phase delay applied to the laser and the phase diagram.

[0021] The intersection of the split phase map and the aliasing intensity map is used to obtain the multicolor imaging result.

[0022] According to the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application, the detector includes four detectors operating in linear array mode. The method of using the detectors to probe the surface of the fluorescent sample to be imaged in a strip scanning manner includes:

[0023] The detector is used to detect the fluorescence signal in the region of the fluorescent sample surface illuminated by the line spot in a direction parallel to the line spot.

[0024] Secondly, this application provides a single-detector single-scan phase-multiplexed multicolor imaging device, comprising:

[0025] The projection module is used to project a line spot of light onto the surface of the fluorescent sample to be imaged using a laser;

[0026] The detection module is used to detect the surface of the fluorescent sample to be imaged using a detector in a strip scanning manner to obtain multi-layer fluorescence signals. During the strip scanning process, the change period is determined based on the single exposure time of the detector, the intensity of the laser is adjusted to change sinusoidally with the change period, and a time delay of a preset wavelength is applied to the intensity change waveform of the laser to generate a phase with a wavelength label.

[0027] An imaging module is used to process the multilayer fluorescence signals to obtain multicolor imaging results.

[0028] Thirdly, this application provides an electronic device, comprising: at least one memory for storing a program; and at least one processor for executing the program stored in the memory, wherein when the program stored in the memory is executed, the processor is configured to execute the single-detector single-scan phase-multiplexed multicolor imaging method described in the first aspect or any possible implementation thereof.

[0029] Fourthly, this application provides a computer-readable storage medium storing a computer program that, when run on a processor, causes the processor to perform the single-detector single-scan phase-multiplexed multicolor imaging method described in the first aspect or any possible implementation of the first aspect.

[0030] Fifthly, this application provides a computer program product that, when run on a processor, causes the processor to execute the single-detector single-scan phase-multiplexed multicolor imaging method described in the first aspect or any possible implementation of the first aspect.

[0031] It is understood that the beneficial effects of the second to fifth aspects mentioned above can be found in the relevant descriptions in the first aspect mentioned above, and will not be repeated here.

[0032] Overall, the technical solutions conceived in this application have the following beneficial effects compared with the prior art:

[0033] By using a detector to perform a single scan of the fluorescent sample surface in a strip scanning manner, and simultaneously adjusting the laser intensity to sinusoidally vary with a period of four times the detector's single exposure time, a phase delay is applied to the laser based on its wavelength. During sample scanning, time and space are simultaneously modulated. This allows for the acquisition of the data required for phase-multiplexed multicolor imaging in a single scan, increasing system imaging throughput and reducing the time consumption of multicolor imaging methods. Furthermore, operation is performed on the original image from online confocal imaging, preserving optical tomography capabilities, and can be combined with the line scanning method, which has the highest scanning throughput, making it ideal for large-scale sample imaging. Attached Figure Description

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

[0035] Figure 1 This is a schematic flowchart of the single-detector single-scan phase multiplexing multicolor imaging method provided in the embodiments of this application;

[0036] Figure 2 This is a system optical path diagram of the single-scan phase multiplexing multicolor imaging method provided in the embodiments of this application;

[0037] Figure 3 This is a schematic diagram illustrating the principle of laser modulation provided in the embodiments of this application;

[0038] Figure 4 This is a schematic diagram of the acquisition of multi-layer fluorescence signals in a single scan, provided in an embodiment of this application.

[0039] Figure 5 This is a schematic diagram of the rearrangement of multilayer fluorescence signals provided in an embodiment of this application;

[0040] Figure 6 This is a flowchart of phase multiplexing multicolor imaging demixing provided in an embodiment of this application;

[0041] Figure 7 This is a schematic diagram of the structure of the single-detector single-scan phase-multiplexed multicolor imaging device provided in the embodiments of this application;

[0042] Figure 8 This is a schematic diagram of the structure of the electronic device provided in the embodiments of this application.

[0043] In all the accompanying drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0044] 1 is the laser for channel 1; 2 is the laser for channel 2; 3 is a dichroic mirror; 4 is a reflecting mirror; 5 is beam expander 1; 6 is beam expander 2; 7 is a cylindrical lens; 8 is an illumination tube lens; 9 is a dichroic mirror; 10 is an objective lens; 11 is a sample; 12 is a motorized translation stage; 13 is a detection tube lens; 14 is an emission filter; 15 is a detector. Detailed Implementation

[0045] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0046] In this article, the term "and / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, or B existing alone. The symbol " / " in this article indicates that the related objects are in an "or" relationship; for example, A / B means A or B.

[0047] In the embodiments of this application, the terms "exemplary" or "for example" are used to indicate that something is an example, illustration, or description. Any embodiment or design that is described as "exemplary" or "for example" in the embodiments of this application should not be construed as being more preferred or advantageous than other embodiments or design. Specifically, the use of the terms "exemplary" or "for example" is intended to present the relevant concepts in a specific manner.

[0048] In the description of the embodiments of this application, unless otherwise stated, "multiple" means two or more, for example, multiple processing units means two or more processing units, multiple elements means two or more elements, etc.

[0049] Next, combined Figures 1-6 The single-scan phase multiplexing multicolor imaging method provided in the embodiments of this application is described.

[0050] Figure 1 This is a schematic flowchart of the single-scan phase-multiplexed multicolor imaging method provided in the embodiments of this application, as shown below. Figure 1 As shown, the method includes the following steps:

[0051] Step 100: Use a laser to project a line spot onto the surface of the fluorescent sample to be imaged;

[0052] Alternatively, a laser beam can be emitted, and a cylindrical lens can be used to form a line spot of a certain width. The line spot is then projected onto the surface of the fluorescent sample to be imaged through an illumination objective and a tube lens.

[0053] Figure 2 This is a system optical path diagram of the single-scan phase-multiplexed multicolor imaging method provided in the embodiments of this application, such as... Figure 2 As shown, in one embodiment of this application, an optical imaging system is used to detect the surface of a fluorescent sample to be imaged. The light emitted from lasers 1 and 2 is combined by a dichroic mirror 3, enters a beam expander system composed of lenses 5 and 6, and then enters a cylindrical lens 7, where it is focused into a line spot. Subsequently, it is projected onto the sample surface through a focusing system composed of a cemented doublet lens 8 and an objective lens 10.

[0054] Step 110: Use a detector to scan the surface of the fluorescent sample to be imaged in a strip scanning manner to obtain multi-layer fluorescence signals. During the strip scanning process, determine the change period based on the single exposure time of the detector, adjust the intensity of the laser to change sinusoidally with the change period, and apply a time delay of a preset wavelength to the intensity change waveform of the laser to generate a phase with wavelength marking.

[0055] Alternatively, the detector can be a linear array detector or a dot array detector.

[0056] Alternatively, if a linear array detector is used, the linear array detector contains 2048 sub-linear arrays, and 4 lines, 8 lines, or 16 lines can be used to detect the surface of the fluorescent sample to be imaged.

[0057] like Figure 2 As shown, the objective lens 10 and the tube lens 13 constitute an infinity correction system that images the fluorescence signal transmitted through the filter 14 onto the surface of the detector 15, enabling the detector to detect the fluorescence signal.

[0058] While the detector performs strip scanning, the illumination laser is modulated so that its intensity varies sinusoidally with the camera frequency. The period of this intensity variation can be a multiple of four, eight, or twelve times the camera's exposure period, effectively broadening the phase spectrum. Simultaneously, a preset wavelength time delay is applied to the laser's intensity waveform, generating a phase with wavelength markings to differentiate color channels. Thus, as the sample moves, the laser also exhibits a periodic, regular variation within the 0-T timeframe.

[0059] In one embodiment of this application, the period is selected to be four times the single exposure time of the detector, that is, the intensity of the laser is adjusted to vary sinusoidally with a period of four times the single exposure time of the detector.

[0060] To ensure the successful execution of phase-multiplexed multicolor imaging, precise synchronization between components is crucial. This includes synchronization between detector imaging and laser intensity modulation, as well as synchronization between linear array detector imaging and translation stage movement during line scanning. To precisely control the timing of these devices, the translation stage is selected as the primary control element.

[0061] Figure 3 This is a schematic diagram illustrating the principle of laser modulation provided in the embodiments of this application, as shown below. Figure 3 As shown, when the translation stage reaches the predetermined position and begins stable operation, it outputs a synchronization signal, which triggers the detector to take continuous pictures. The detector sends a feedback signal at the start of each exposure, which is used as a synchronization signal between the camera and the NI acquisition card. Based on this feedback signal, the NI acquisition card outputs a voltage waveform with a specific frequency and fixed delay, thereby achieving direct modulation of the laser intensity.

[0062] Optionally, the single-detector single-scan phase-multiplexed multicolor imaging method provided in this application can perform three-dimensional imaging, such as imaging by directly lifting samples from thin slices and light-transparent samples, or imaging by cutting and lifting samples from complete tissues and organs. The multi-layer fluorescence signals acquired by the detector can include three-dimensional fluorescence signal data illuminated by the waist of the Gaussian beam and three-dimensional fluorescence signal data illuminated away from the waist of the beam.

[0063] Figure 4 This is a schematic diagram of the acquisition of multi-layer fluorescence signals in a single scan according to an embodiment of this application, as shown below. Figure 4As shown, in one embodiment of this application, the fluorescence signal of the fluorescent sample to be imaged is obtained by scanning the entire sample range using a translation stage. Specifically, firstly, the fluorescence signal of the area brightened by the line spot on the surface of the fluorescent sample to be imaged is detected by a detector to obtain a single-layer fluorescence signal, including spatial coding data and color time coding data. Then, the fluorescent sample to be imaged is lifted by the translation stage, and the fluorescence signal of the area brightened by the line spot on the surface of the fluorescent sample to be imaged is detected again by the detector to obtain a single-layer fluorescence signal. The above steps are repeated until the fluorescent sample to be imaged is completely detected, thereby obtaining a multi-layer fluorescence signal.

[0064] The above spatiotemporal multiplexing process can simultaneously obtain spatially encoded data and color-time encoded data.

[0065] Step 120: Process the multi-layer fluorescence signal to obtain multicolor imaging results.

[0066] After acquiring multi-layer fluorescence signal data in a single scan, the data is processed to obtain multicolor imaging results.

[0067] The single-scan phase-multiplexed multicolor imaging method provided in this application performs a single scan of the fluorescent sample surface to be imaged using a detector in a strip scanning manner. Simultaneously, the laser intensity is adjusted to sinusoidally vary with a period of four times the detector's single exposure time. A phase delay is applied to the laser based on its wavelength, resulting in simultaneous temporal and spatial modulation during sample scanning. This allows for the acquisition of the data required for phase-multiplexed multicolor imaging in a single scan, increasing system imaging throughput and reducing the time consumption of multicolor imaging methods. Furthermore, since the operation is performed on the original image from online confocal imaging, optical tomography capabilities are preserved, and it can be combined with the line scanning method, which boasts the highest scanning throughput, making it highly suitable for large-scale sample imaging.

[0068] In some embodiments, the detector includes multiple sub-detectors. During the process of using the detector to scan the surface of the fluorescent sample to be imaged in a strip scanning manner, the multiple sub-detectors in the detector record the fluorescence signals of different sample points under the same light intensity at the same time, and finally record the fluorescence signals of different sample points under different light intensity.

[0069] Within the illumination range, multiple sub-detectors can be used to record the signal of the sample simultaneously. In this way, the signals of different sample points under the same light intensity are recorded at the same time. As the sample passes through each sub-detector in turn, the different sub-detectors record the signals of different sample points under different light intensity in turn.

[0070] This strategy allows a single camera to acquire all the data of a demodulated monochrome image in a single scan, without any additional pauses or the time required for multiple scans.

[0071] In some embodiments, step 120 specifically includes:

[0072] Step 1201: Process the multilayer fluorescence signal to obtain an intensity map of all fluorescence channels mixed together and a phase map that can be used to distinguish the fluorescence channels;

[0073] Step 1202: Based on the intensity map and phase map, the fluorescence channels are split to obtain multicolor imaging results.

[0074] By processing multi-layer fluorescence signals, which include multi-color data encoded in both time and space, an intensity map of mixed color channels and a phase map of distinguishable colors are obtained. After splitting the fluorescence channels, multi-color imaging results with improved signal-to-background ratio and imaging throughput can be obtained.

[0075] Optionally, the multilayer fluorescence signal can be processed by a four-step phase-shift solution, a linear solution, or deep learning to obtain an intensity map of all fluorescence channels and a phase map that can be used to distinguish the fluorescence channels. This application does not limit this.

[0076] In some embodiments, step 1201 specifically includes:

[0077] Step 12011: Rearrange the multilayer fluorescence signals according to the time periodic pattern to generate four images taken within one period;

[0078] Step 12012: Based on the four images, use the four-step phase-shifting method to solve for the intensity map of all fluorescence channels and the phase map that can be used to distinguish the fluorescence channels.

[0079] Optical imaging systems apply specific phase delays φ1-φ4 to different lasers. For a given point in space, that point may be excited by one or more of these lasers. The emitted fluorescence has a phase of φ0, and its intensity variation follows the following formula:

[0080]

[0081] in, , representing the fluorescence intensity distribution The DC component, For focal plane information, , For the sample surface coordinates, For modulation frequency, For time.

[0082] To solve this phase related to the excitation light The phase of the fluorescence signal is rearranged according to a time periodic pattern to generate four images captured within one period, which can be expressed as:

[0083]

[0084] in, , , , Four images taken within one period. The modulation period.

[0085] Further, using the four-step phase-shifting method, an intensity map of all fluorescence channels and a phase map that can be used to distinguish fluorescence channels are obtained, which can be expressed as:

[0086]

[0087] Figure 5 This is a schematic diagram of the rearrangement of multilayer fluorescence signals provided in an embodiment of this application, as shown below. Figure 5 As shown, the four-step phase-shifting method is used to solve for the intensity aliasing images of all fluorescence channels and the phase images that can distinguish the fluorescence channels.

[0088] Phase After the phase, it can be based on The fluorescence channels were split.

[0089] In particular, this application has a certain robustness to excitation crosstalk and spatially overlapping multicolor samples. It can distinguish excitation crosstalk by phase peak shift and use linear demixing in the phase map to demix spatial colocation information.

[0090] In some embodiments, step 1202 specifically includes:

[0091] Step 12021: Binarize the intensity map, multiply the obtained mask with the phase map to obtain the aliasing intensity map;

[0092] Step 12022: Decompose the multiplexed channel based on the phase delay and phase diagram applied to the laser;

[0093] Step 12023: Take the intersection of the split phase map and the aliasing intensity map to obtain the multicolor imaging result.

[0094] Figure 6 This is a flowchart of phase multiplexing multicolor imaging demixing provided in an embodiment of this application, such as... Figure 6 As shown, when splitting different channels, the aliasing intensity map is first binarized, and the resulting mask is multiplied by the phase image to reduce the influence of the background phase on channel splitting. Next, the multiplexed channels are split based on the phase delay applied to the laser and the phase map. Then, the intersection of the split phase map and the aliasing intensity map is taken to obtain the final single-channel intensity map, achieving phase-multiplexed multicolor imaging.

[0095] The single-detector, single-scan phase-multiplexed multicolor imaging method provided in this application can significantly reduce the scanning time of multiple phase-multiplexed multicolor imaging methods, reducing the time by several times compared to multicolor imaging methods that require changing filters. Furthermore, this method operates on the original image obtained through online confocal imaging, preserving optical tomography capabilities, and can be combined with the line scanning method, which boasts the highest scanning throughput, making it highly suitable for imaging large-scale samples.

[0096] The single-detector single-scan phase-multiplexed multicolor imaging method provided in this application can effectively distinguish fluorescent dyes with excitation light crosstalk. At the same time, for situations where multiple substances emit light in a space, the intensity of different colors of fluorescence can be separated by using the phase linear mixing law.

[0097] In some embodiments, the detector includes four detectors operating in linear array mode, and step 120 specifically includes:

[0098] The detector is used to detect the fluorescence signal in the area of ​​the fluorescent sample surface illuminated by the line spot in a direction parallel to the line spot.

[0099] This application uses a linear array detector comprising four operating in linear array mode, wherein the detected single-layer fluorescence signal includes the raw data from multi-line array detection: fluorescence sample signal detected by linear array detector 1, fluorescence sample signal detected by linear array detector 2, fluorescence sample signal detected by linear array detector 3, and fluorescence sample signal detected by linear array detector 4.

[0100] The detector operates in linear array mode, detecting fluorescence signals in the bright areas illuminated by the linear light spots, parallel to the direction of the light spots.

[0101] Figure 7 This is a schematic diagram of the structure of the single-detector single-scan phase-multiplexed multicolor imaging device provided in the embodiments of this application, as shown below. Figure 7 As shown, the device includes a projection module 710, a detection module 720, and an imaging module 730, wherein:

[0102] Projection module 710 is used to project a line spot onto the surface of the fluorescent sample to be imaged using a laser;

[0103] The detection module 720 is used to detect the surface of the fluorescent sample to be imaged in a strip scanning manner using a detector to obtain multi-layer fluorescence signals. During the strip scanning process, the change period is determined based on the single exposure time of the detector, the intensity of the laser is adjusted to change sinusoidally with the change period, and a time delay of a preset wavelength is applied to the intensity change waveform of the laser to generate a phase with a wavelength label.

[0104] The imaging module 730 is used to process multi-layer fluorescence signals to obtain multi-color imaging results.

[0105] It should be understood that the above-described device is used to execute the methods in the above embodiments. The implementation principle and technical effect of the corresponding program modules in the device are similar to those described in the above methods. The working process of the device can be referred to the corresponding process in the above methods, and will not be repeated here.

[0106] Based on the methods in the above embodiments, Figure 8 An example is a schematic diagram of the physical structure of an electronic device, such as... Figure 8 As shown in the illustration, this application provides an electronic device that may include a processor 810, a communications interface 820, a memory 830, and a communication bus 840. The processor 810, communications interface 820, and memory 830 communicate with each other via the communication bus 840. The processor 810 can call logical instructions stored in the memory 830 to execute the single-detector single-scan phase-multiplexed multicolor imaging method described in the above embodiment.

[0107] Furthermore, the logic instructions in the aforementioned memory 830 can be implemented as software functional units and, when sold or used as independent products, can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of this application, in essence, or the part that contributes to the prior art, or a portion of the technical solution, can be embodied in the form of a software product. This computer software product is stored in a storage medium and includes several instructions to cause a computer device (which may be a personal computer, server, or network device, etc.) to execute all or part of the steps of the single-detector single-scan phase-multiplexed multicolor imaging method described in the various embodiments of this application.

[0108] Based on the methods in the above embodiments, this application provides a computer-readable storage medium storing a computer program. When the computer program runs on a processor, it causes the processor to execute the single-detector single-scan phase multiplexing multicolor imaging method in the above embodiments.

[0109] Based on the methods in the above embodiments, this application provides a computer program product that, when running on a processor, causes the processor to execute the single-scan phase multiplexing multicolor imaging method in the above embodiments.

[0110] It is understood that the processor in the embodiments of this application can be a central processing unit (CPU), or other general-purpose processors, digital signal processors (DSPs), application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), or other programmable logic devices, transistor logic devices, hardware components, or any combination thereof. A general-purpose processor can be a microprocessor or any conventional processor.

[0111] The method steps in this application embodiment can be implemented in hardware or by a processor executing software instructions. The software instructions can consist of corresponding software modules, which can be stored in random access memory (RAM), flash memory, read-only memory (ROM), programmable read-only memory (PROM), erasable programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), registers, hard disks, portable hard disks, CD-ROMs, or any other form of storage medium known in the art. An exemplary storage medium is coupled to the processor, enabling the processor to read information from and write information to the storage medium. Of course, the storage medium can also be a component of the processor. The processor and the storage medium can reside in an ASIC.

[0112] In the above embodiments, implementation can be achieved entirely or partially through software, hardware, firmware, or any combination thereof. When implemented using software, it can be implemented entirely or partially as a computer program product. The computer program product includes one or more computer instructions. When the computer program instructions are loaded and executed on a computer, all or part of the processes or functions described in the embodiments of this application are generated. The computer can be a general-purpose computer, a special-purpose computer, a computer network, or other programmable device. The computer instructions can be stored in a computer-readable storage medium or transmitted through the computer-readable storage medium. The computer instructions can be transmitted from one website, computer, server, or data center to another website, computer, server, or data center via wired (e.g., coaxial cable, fiber optic, digital subscriber line (DSL)) or wireless (e.g., infrared, wireless, microwave, etc.) means. The computer-readable storage medium can be any available medium that a computer can access or a data storage device such as a server or data center that integrates one or more available media. The available medium can be a magnetic medium (e.g., floppy disk, hard disk, magnetic tape), an optical medium (e.g., DVD), or a semiconductor medium (e.g., solid-state disk (SSD)).

[0113] It is understood that the various numerical designations used in the embodiments of this application are merely for the convenience of description and are not intended to limit the scope of the embodiments of this application.

[0114] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application should be included within the scope of protection of this application.

Claims

1. A single-detector, single-scan, phase- multiplexed, multichrome imaging method, characterized by, The method comprises the steps of: projecting a line spot onto a fluorescent sample surface to be imaged using a laser; detecting the fluorescent sample surface to be imaged in a strip scanning manner using a detector to obtain multi-layer fluorescent signals, and determining a variation period based on a single exposure time of the detector during the strip scanning process, adjusting the intensity of the laser to vary in a sinusoidal manner with the variation period, imposing a time delay of a preset wavelength on the intensity variation waveform of the laser to generate a phase with wavelength identification; processing the multi-layer fluorescent signals to obtain a multi-color imaging result; The processing of the multi-layer fluorescent signals to obtain a multi-color imaging result comprises the steps of: processing the multi-layer fluorescent signals to obtain an intensity map of all fluorescent channels mixed and a phase map that can be used to distinguish fluorescent channels; splitting the fluorescent channels based on the intensity map and the phase map to obtain a multi-color imaging result; The processing of the multi-layer fluorescent signals to obtain an intensity map of all fluorescent channels mixed and a phase map that can be used to distinguish fluorescent channels comprises the steps of: rearranging the multi-layer fluorescent signals in a time period to generate four images taken within a period; solving the intensity map of all fluorescent channels mixed and the phase map that can be used to distinguish fluorescent channels using a four-step phase shift method based on the four images.

2. The single-detector, single-scan, phase-encoded, multichromatic imaging method of claim 1, wherein, The detector comprises a plurality of sub-detectors, and during the detection of the fluorescent sample surface to be imaged in a strip scanning manner using the detector, the plurality of sub-detectors in the detector record fluorescent signals of different sample points under the same light intensity modulation at the same time, and finally record fluorescent signals of different sample points under different light intensity modulations.

3. The single-detector, single-scan, phase-encoded, multichromatic imaging method of claim 1, wherein, The detector comprises four detectors working in a line array mode, and the detection of the fluorescent sample surface to be imaged in a strip scanning manner using the detector comprises the steps of: detecting fluorescent signals of the area of the fluorescent sample surface to be imaged illuminated by the line spot in a direction parallel to the line spot using the detector.

4. A single-detector, single-scan, phase- multiplexed, multichromatic imaging apparatus, characterized by, The method comprises the steps of: projecting a line spot onto a fluorescent sample surface to be imaged using a laser; detecting the fluorescent sample surface to be imaged in a strip scanning manner using a detector to obtain multi-layer fluorescent signals, and determining a variation period based on a single exposure time of the detector during the strip scanning process, adjusting the intensity of the laser to vary in a sinusoidal manner with the variation period, imposing a time delay of a preset wavelength on the intensity variation waveform of the laser to generate a phase with wavelength identification; processing the multi-layer fluorescent signals to obtain a multi-color imaging result; The processing of the multi-layer fluorescent signals to obtain a multi-color imaging result comprises the steps of: processing the multi-layer fluorescent signals to obtain an intensity map of all fluorescent channels mixed and a phase map that can be used to distinguish fluorescent channels; splitting the fluorescent channels based on the intensity map and the phase map to obtain a multi-color imaging result; The processing of the multi-layer fluorescent signals to obtain an intensity map of all fluorescent channels mixed and a phase map that can be used to distinguish fluorescent channels comprises the steps of: rearranging the multi-layer fluorescent signals in a time period to generate four images taken within a period; solving the intensity map of all fluorescent channels mixed and the phase map that can be used to distinguish fluorescent channels using a four-step phase shift method based on the four images. Based on the four images, the intensity map of all the fluorescent channels mixed and the phase map that can be used to distinguish the fluorescent channels are solved using a four-step phase shifting method.

5. An electronic device, comprising: The method comprises: at least one memory for storing a computer program; at least one processor for executing the program stored in the memory, and when the program stored in the memory is executed, the processor is configured to execute the single-detector single-scan phase multiplexing multi-color imaging method according to any one of claims 1-3.

6. A computer-readable storage medium storing a computer program, the computer program comprising instructions that, when executed by a computer, cause the computer to perform the method of any one of claims 1 to 5. When the computer program runs on the processor, the processor is caused to execute the single-detector single-scan phase multiplexing multi-color imaging method according to any one of claims 1-3.

7. A computer program product, characterised in that, When the computer program product runs on the processor, the processor is caused to execute the single-detector single-scan phase multiplexing multi-color imaging method according to any one of claims 1-3.

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