Imaging control method of multi-photon imaging equipment and multi-photon imaging system

Abstract

The invention provides an imaging control method of multi-photon imaging equipment and a multi-photon imaging system, and relates to the technical field of optics. In order to solve the problem of cable torsion in an imaging system, a photoelectric composite slip ring is arranged in the imaging equipment, so that the imaging equipment rotates based on the rotation of an imaging probe relative to the photoelectric composite slip ring; the cable torsion caused by the rotation of the imaging probe (or the imaging object) is eliminated, so that the imaging probe and the imaging object can relatively freely move in the imaging space. Meanwhile, in consideration of the problem that the excitation light power carried by the excitation light optical fiber is unstable due to the fact that the coupling relation of the excitation light optical fiber at the photoelectric composite slip ring is possibly unstable when the excitation light optical fiber rotates, an excitation light detection device is arranged between the optical slip ring rotor end and the excitation light path so as to collect the actual power information of the excitation light; therefore, the laser for exciting light and / or the processor for imaging are / is compensated based on the actual power information, so that the stability of the imaging process is maintained.

Description

Technical Field

[0001] The present application relates to the field of optical technology, and in particular to an imaging control method for a multi-photon imaging device and a multi-photon imaging system. Background Art

[0002] Multiphoton imaging is an advanced microscopy technique that exploits the multiphoton effect. It achieves high-resolution imaging of deep tissue by simultaneously absorbing multiple low-energy photons (such as two-photons or three-photons) to excite fluorescent molecules. Multiphoton imaging often requires an imaging probe that modulates the excitation light. During imaging, the probe is in direct contact with the object, triggering the multiphoton phenomenon in the object and collecting the fluorescence signal.

[0003] In actual detection, multiphoton imaging is a point effect, and scanning imaging is often required, which takes a long time. Rotation of the imaging object or imaging probe can cause cable twisting, affecting detection results. For example, when imaging equipment is used in biological experiments and the imaging object is a living animal such as a mouse or rabbit, the object's own motion (such as self-rotation) can cause cable twisting, affecting the motion of the imaging object.

[0004] Therefore, how to eliminate the cable twisting caused by the imaging probe is a technical problem that needs to be solved urgently by those skilled in the art. Summary of the Invention

[0005] In view of this, an embodiment of the present application provides an imaging control method and a multiphoton imaging system for a multiphoton imaging device, which actively releases the cable torsion caused by the rotation of the imaging probe (active or passive) by setting an optoelectronic composite slip ring for twisting rotation.

[0006] In a first aspect, the present application provides an imaging control method for a multiphoton imaging device. The multiphoton imaging device includes an imaging host, an imaging probe, an optoelectronic composite slip ring, and an excitation light detection device. The imaging host is configured to release excitation light and generate a fluorescence image based on the returned fluorescence signal. The imaging probe is configured to release excitation light toward the imaging object, triggering a multiphoton fluorescence effect and collecting the fluorescence signal generated by the multiphoton fluorescence effect. The imaging host is connected to the stator end of the optoelectronic composite slip ring, and the imaging probe is connected to the mover end of the optoelectronic composite slip ring. The mover end is coupled to the stator end and driven to rotate relative to the stator end. The optoelectronic composite slip ring includes a smooth ring connected to an excitation light fiber and an electric slip ring connected to at least one metal conductor. The excitation light detection device has one end connected to the smooth ring mover end of the smooth ring and the other end connected to the electric slip ring mover end of the electric slip ring. The device is configured to collect excitation light power information within the excitation light fiber at the smooth ring mover end. The imaging control method includes determining the operating power of the excitation light fiber using the excitation light detection device. In response to relative rotation of the optoelectronic composite slip ring, the excitation light detection device determines the post-rotation power of the excitation light fiber after rotation. The working parameters of the imaging host are adjusted based on the working power and the power after rotation to generate a fluorescent image of the imaging object after the excitation light fiber is rotated.

[0007] In a second aspect, the present application provides a multiphoton imaging system, comprising a multiphoton detection device corresponding to the imaging control method described in the first aspect, a controller for executing the imaging control method described in the first aspect on the multiphoton detection device, and an imaging space accommodating an imaging object. The imaging probe of the multiphoton detection device is connected to the imaging object, and an optoelectronic composite slip ring of the multiphoton detection device is disposed at an edge of the imaging space based on the connection direction between the imaging probe and the imaging object. The optoelectronic composite slip ring is configured to rotate based on the rotation of the imaging probe relative to the optoelectronic composite slip ring. The excitation light detection device of the multiphoton detection device is configured to detect power information of excitation light between the smooth ring rotor end of the optoelectronic composite slip ring and the excitation light path of the imaging probe to adjust the imaging process.

[0008] Therefore, the imaging control method and multiphoton imaging system of the multiphoton imaging device provided by the present application are designed to solve the problem of cable twisting caused by the imaging object in the detection system. The detection device of the present application is provided with an optoelectronic composite slip ring, which rotates based on the rotation of the imaging probe relative to the optoelectronic composite slip ring to eliminate the cable twisting caused by the rotation of the imaging probe (or imaging object) and enable the imaging probe and imaging object to move relatively freely within the imaging space. At the same time, considering that the coupling relationship of the excitation light fiber at the optoelectronic composite slip ring may be unstable when it rotates, thereby causing the problem of unstable excitation light power carried by the excitation light fiber, the present application provides an excitation light detection device between the smooth ring mover end and the excitation light path to collect the actual power information of the excitation light, so as to compensate the excitation light laser and / or the imaging processor based on the actual power information to maintain the stability of the imaging process. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0010] Figure 1 It is a system schematic diagram of a detection system provided by an exemplary embodiment of the present application.

[0011] Figure 2 Schematic diagram of a longitudinal section of an optoelectronic composite slip ring provided by an exemplary embodiment of the present application.

[0012] Figure 3 It is a schematic structural diagram of an excitation light detection device provided by an exemplary embodiment of the present application.

[0013] Figure 4 Schematic diagram of the structure of an imaging probe provided by an exemplary embodiment of the present application.

[0014] Figure 5 is an exemplary flow chart of an imaging control method provided by an exemplary embodiment of the present application.

[0015] Figure 6 is an exemplary flow chart of a rotation control and compensation process provided by an exemplary embodiment of the present application.

[0016] Among them, 10, imaging system; 100, imaging device; 200, imaging space; 300, imaging object; 110, imaging host; 120, imaging probe; 121, collimation module; 122, scanning galvanometer; 123, lens module; 124, objective lens module; 130, optoelectronic composite slip ring; 131, central smooth ring; 132, hollow electric slip ring; 133, drive motor; 140, optoelectronic composite cable; 141, excitation light fiber; 142, fluorescent fiber; 150, excitation light detection device; 151, spectrometer module; 1511, quartz structure; 1512, coupling structure; 152, power detection module; 1521, beam splitting structure; 1522, multiple power sensors; 153, beam splitting fiber. DETAILED DESCRIPTION

[0017] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0018] Application Overview:

[0019] As mentioned above, in a multi-photon imaging device, in order to relieve cable twisting caused by the rotation of the imaging probe (including the rotation generated when the user uses the imaging probe and the rotation caused by the rotation of the imaging object itself), a photoelectric composite slip ring can be used to relieve twisting.

[0020] To further illustrate the cable untwisting process based on the optoelectronic composite slip ring, the present application provides a system schematic diagram of an imaging system provided with an optoelectronic composite slip ring ( Figure 1 ).

[0021] like Figure 1 As shown, the imaging system 10 may include an imaging device 100 and an imaging space 200 , wherein an imaging object 300 is disposed in the imaging space 200 .

[0022] The imaging device 100 may refer to a device that performs imaging using multiphoton phenomena. The imaging device 100 may include, but is not limited to, a multiphoton imaging device, a multi-harmonic imaging device, etc. The specific structure of the imaging device 100 may be found in the subsequent description.

[0023] Taking two-photon imaging as an example, during the actual imaging process, when excitation light is released onto the imaging object 300, two photons in the excitation light converge at a certain depth within the imaging object 300, thereby stimulating the multiphoton phenomenon of the relevant fluorescent dye or structure, thereby releasing a fluorescent signal. The imaging device 100 can form a detection image by changing the imaging position to perform planar scanning and / or depth scanning.

[0024] The imaging space 200 may refer to the space where the imaging object 300 is located, and is generally established based on a physical structure. For example, in a biological experiment, the imaging space 200 may be constructed based on an ecological box that accommodates the experimental object (generally presented as Figure 1 In addition, in practical applications, the imaging space 200 may not be limited to biological experiments. For example, the aforementioned imaging device 100 may also be used in human skin testing, and the imaging space 200 only needs to provide support for the optoelectronic composite slip ring of the imaging device 100 (e.g., a support frame). In addition, when the imaging probe of the imaging device 100 is configured as a handheld probe, the imaging space formed therefrom can also be equivalently understood as a side-opening structure (i.e., the optoelectronic composite cable enters the imaging space from the side).

[0025] The imaging object 300 may refer to the detection object of the aforementioned imaging system 10. Figure 1 A mouse is used as an example to illustrate the detection and detwisting process. In actual applications, the imaging object 300 may also be other living organisms (such as rabbits, humans, etc.) or partial tissues (such as diseased tissues, body sampled tissues, etc.).

[0026] Taking mice as an example, when using the aforementioned multiphoton imaging equipment for detection, images of the mouse brain can generally be detected. Specifically, a window can be opened in the mouse's head and infiltrated with a fluorescent dye, so that some structures in the mouse brain can be excited by the twin light phenomenon and release fluorescence of different wavelengths, forming a detection image. In practical applications, the detection image of the mouse brain generally reflects structures such as calcium ions, mitochondria, and synapses.

[0027] To further illustrate the untwisting process, the aforementioned imaging device 100 further includes an imaging host 110, an imaging probe 120, an optoelectronic composite slip ring 130, and an optoelectronic composite cable 140. The optoelectronic composite slip ring 130 is disposed on the optoelectronic composite cable 140, dividing the optoelectronic composite cable 140 into an optoelectronic composite cable on the imaging host 110 side and an optoelectronic composite cable on the imaging probe 120 side. The two sections of the optoelectronic composite cable are coupled via the optoelectronic composite slip ring 130.

[0028] Based on the aforementioned imaging space 200, the imaging host 110 is disposed outside the imaging space 200, the optoelectronic composite slip ring 130 is disposed on the imaging space 200, and the imaging probe 120 is disposed on the imaging object 300 within the imaging space 200. The optoelectronic composite cable 140 is connected between the imaging host 110 and the imaging probe 120 via the optoelectronic composite slip ring 130.

[0029] The imaging host 110 may be formed by components disposed outside the imaging space 200 , and its main functions may be to release excitation light, generate timing control signals, and generate detection images based on fluorescence signals. It may generally include components such as a laser and a controller.

[0030] The aforementioned imaging probe 120 may refer to a device that contacts the imaging object 300 , and generally needs to form an excitation light path for releasing excitation light to the imaging object 300 and a fluorescence light path for collecting fluorescence optical signals.

[0031] The optoelectronic composite slip ring 130 can be a slip ring assembly capable of simultaneously coupling and rotating optical fibers and metal conductors. The optoelectronic composite slip ring 130 can include a stator end and a mover end. The mover end can be driven (e.g., by a motor) to rotate relative to the stator end. During the relative rotation of the mover end and the stator end, the cables connected to the two ends can maintain a coupled state (i.e., the metal conductors maintain an electrical connection, and the optical fibers remain optically coupled).

[0032] Based on the aforementioned stator and mover ends, the optoelectronic composite cable 140 on the imaging host 110 side can be connected to the stator end, maintaining the stability of the cable between the imaging host 110 and the optoelectronic composite slip ring 130. The optoelectronic composite cable 140 on the imaging probe 120 side can be connected to the mover end and rotate accordingly when the imaging probe 120 rotates, thereby relieving the torsion of the optoelectronic composite cable 140 on the imaging probe 120 side caused by the imaging probe 120.

[0033] Furthermore, in actual applications, the present applicant has discovered that, to achieve the formation of a detection image, the imaging host 110 often needs to receive a fluorescence signal. Simultaneously, the imaging host 110 needs to release excitation light. Therefore, the optoelectronic composite slip ring 130 needs to simultaneously detwist the fluorescence fiber and the excitation light fiber. To simplify the fluorescence fiber, optoelectronic composite slip ring 130 can perform photoelectric conversion of the fluorescence signal between the optoelectronic composite slip ring 130 and the imaging probe 120. This allows the fluorescence signal transmitted to the imaging host 110 to be transmitted via a metal conductor, while the optoelectronic composite slip ring 130 only needs to detwist the excitation light fiber. The specific optoelectronic conversion process and its device configuration can be found in related art and will not be detailed here.

[0034] As mentioned above, in nonlinear optical detection equipment, as the imaging requirements change, its actual excitation light fiber may need to simultaneously transport excitation light of multiple wavelengths. In order to achieve the transmission of multi-wavelength excitation light, the common solid optical fiber has limited spectral bandwidth, and the detection equipment often requires a highly stable and high-power laser, while the optical fiber can usually only transmit a single wavelength of laser. The detection equipment (such as the detection equipment shown in the aforementioned CN119147464A) can use a multimode photonic crystal fiber for excitation light transmission, so that the multimode photonic crystal fiber can transmit multiple wavelengths of excitation light within the target wavelength range.

[0035] In the field of multiphoton imaging, the aforementioned multimode photonic crystal fiber can be further configured as a hollow-core antiresonant fiber (HC-ARF). This fiber has a two-layer structure: a silica core surrounded by a cladding containing multiple equally spaced air holes. The cladding, which forms the air holes, can form the aforementioned resonant cavity structure, thereby strongly confining the excitation light within the target bandwidth.

[0036] Specifically, in multi-photon imaging, the excitation light is generally 550nm to 1800nm, and the aforementioned multi-mode photonic crystal fiber can be designed to have good performance within the range of 550nm to 1800nm ​​to transmit excitation light beams of multiple wavelengths.

[0037] Therefore, in an actual device, the optoelectronic composite cable 140 may include a multimode photonic crystal fiber and at least one metal conductor, and the multimode photonic crystal fiber transmits multiple wavelengths of excitation light within the target wavelength range. The optical fiber is connected to the smooth ring in the optoelectronic composite slip ring 130 corresponding to the multimode photonic crystal fiber, so that the rotation of the smooth ring enables relative rotation of the multimode photonic crystal fibers at both ends.

[0038] However, during the actual untwisting process, the alignment accuracy and stability of the optoelectronic composite slip ring 130 itself are limited, and the properties of the excitation light beam may change before and after the optoelectronic composite slip ring 130 rotates. For example, the rotation of the optoelectronic composite slip ring 130 may cause the alignment between the stator and mover ends to change, or the shape of the mover end to change. This may cause the actual optical path to differ before and after, leading to fluctuations in the power of the excitation light.

[0039] It should be noted that during multiphoton imaging, the excitation light emitted by the laser will experience some loss as it moves toward the focal position of the imaging object. However, this loss should be known and can be compensated for by adjusting device parameters such as the excitation power and the fluorescence sensor threshold. However, the rotatability of the aforementioned optoelectronic composite slip ring, combined with the complexity of the actual operating environment, may make the loss (or gain) caused by its rotation unpredictable, seriously affecting the imaging effect (e.g., possible failure to excite fluorescence or abnormal focus range).

[0040] Therefore, in order to understand the impact of the rotation of the optoelectronic composite slip ring 130 on the excitation light, the multiphoton imaging device 100 provided in this application can also be provided with an excitation light detection device 150. One end of the excitation light detection device 150 is connected to the smooth ring mover end of the smooth ring, and the other end is connected to the electric slip ring mover end of the electric slip ring, and is used to collect power information of the excitation light in the excitation light fiber at the smooth ring mover end.

[0041] That is, one end of the aforementioned excitation light detection device 150 can be coupled between the smooth ring mover end of the smooth ring and the excitation light path, so that the excitation light beam is split and enters the excitation light detection device 150 for detection (such as power detection), thereby determining its power information and connecting it to the electric slip ring mover end through a metal wire and transmitting it to the imaging host 110 through the optoelectronic composite slip ring 130, so that the imaging host 110 can perform imaging control based on the power information to execute the imaging control method provided by this application. For more information about the imaging control method, please refer to Figure 5 、 6 and its related descriptions.

[0042] Based on the aforementioned multi-photon imaging device and system, an optoelectronic composite slip ring is provided to solve the problem of cable twisting caused by the imaging object in the detection system. That is, the imaging probe can rotate based on the rotation of the optoelectronic composite slip ring to eliminate the cable twisting caused by the rotation of the imaging probe (or imaging object), so that the imaging probe and the imaging object can move relatively freely within the imaging space. At the same time, considering that the coupling relationship of the excitation light fiber at the optoelectronic composite slip ring may be unstable when the excitation light fiber rotates, thereby causing the problem of unstable excitation light power carried by the excitation light fiber, the present application provides an excitation light detection device between the smooth ring mover end and the excitation light path to collect the actual power information of the excitation light, so as to compensate the excitation light laser and / or the imaging processor based on the actual power information to maintain the stability of the imaging process.

[0043] It should be noted that the aforementioned optoelectronic composite cable 140 on the imaging host 110 side and the optoelectronic composite cable 140 on the imaging probe 120 side merely refer to the cables between the aforementioned devices as a whole, and do not require that the optoelectronic composite cable 140 on the imaging host 110 side and the optoelectronic composite cable 140 on the imaging probe 120 side be bundled together to form a single composite cable. For example, to reduce the restrictions on the movement range of the individual cables in the optoelectronic composite cable 140 on the imaging probe 120 side, the individual optical fibers and metal conductors within the optoelectronic composite cable 140 on the imaging probe 120 side can be separated and independently connected between the imaging probe 120 and the optoelectronic composite slip ring 130.

[0044] Exemplary configuration of the excitation light detection device:

[0045] In order to further illustrate the setting of the excitation light detection device in combination with the structure of the optoelectronic composite slip ring, the present application provides a cross-sectional schematic diagram of the optoelectronic composite slip ring ( Figure 2 ). The cross section can be viewed at the center of the optoelectronic composite slip ring along the axial direction of the optoelectronic composite slip ring.

[0046] like Figure 2 As shown, the optoelectronic composite slip ring 130 may include a central smooth ring 131 and a hollow electric slip ring 132. The central smooth ring 131 is arranged at the rotation axis of the hollow electric slip ring 132, and the mover end of the central smooth ring 131 is connected to the mover end of the hollow electric slip ring 132 to rotate synchronously.

[0047] In order to realize the rotation of the aforementioned central smooth ring 131 and the mover end of the hollow electric slip ring 132, the optoelectronic composite slip ring 130 may further include a drive motor 133, which can be connected to the mover end of the hollow electric slip ring 132 through transmission (such as through a gear transmission connection) to drive the mover end of the hollow electric slip ring 132 to rotate.

[0048] Based on the above-mentioned setting, the central smooth ring 131 only involves the coupling of the excitation light fiber (that is, the stator end of the central smooth ring is connected to the excitation light fiber 141 in the optoelectronic composite cable 140 on the imaging host 110 side, and the mover end is connected to the excitation light fiber 141 in the optoelectronic composite cable 140 on the imaging probe 120 side). Then, the mover end and the stator end of the central smooth ring 131 can be coupled inside the central smooth ring 131 through the beam alignment module to realize the excitation light transmission during the rotation process.

[0049] The aforementioned hollow electric slip ring 132 may include multiple coaxial channels, and the same channels at the stator end and the mover end of the hollow electric slip ring 132 may be electrically connected (eg, through contacts, brushes, etc.) to achieve electrical connection during rotation.

[0050] Furthermore, considering that the aforementioned excitation light is actually configured as multiple excitation lights of different wavelengths within the target wavelength range, the aforementioned excitation light fiber 141 can be further configured as a photonic crystal fiber for transmitting multiple excitation lights of different wavelengths within the target wavelength range.

[0051] To detect the excitation light within the photonic crystal fiber, the excitation light detection device 150 includes a spectrometer module 151 and a power detection module 152. The spectrometer module 151 can split the excitation light from the excitation light fiber 141 / excitation light path on the imaging probe 120 side and transmit it to the power detection module 152. The power detection module 152 then detects the power of the excitation light to determine the power information of the excitation light.

[0052] That is, the aforementioned spectrometer module 151 has an input end, an output end and a spectrometer end, wherein the input end is connected to the smooth ring mover end, the output end is connected to the excitation light path of the imaging probe 120, and the spectrometer end is used to split the excitation light from the smooth ring mover end and the excitation light path.

[0053] The power detection module 152 has an input end and an output end. The input end is coupled to the light splitting end of the light splitting module 151 , and the output end can transmit the power information of the split excitation light to the imaging host 110 .

[0054] In practical applications, the present application does not limit the actual structure of the light splitting module 151. For example, the excitation light beam splitting can be achieved based on diffraction optical element (DOE) technology, grating structure design, spatial optics, and refraction and reflection of tilted light mirrors. It only needs to ensure that the split laser beam can reflect the excitation light intensity in the excitation light path. For example, the two can be linearly / positively correlated. In addition, the optical properties of the light beam split by the aforementioned light splitting module 151 are generally consistent with the original light beam, that is, the aforementioned light splitting module 151 is generally not implemented by a polarization beam splitter.

[0055] The aforementioned power detection module 152 can generally be constructed based on a laser power detector (PD, Photodetector). A laser power detector is a device that converts optical signals into electrical signals to achieve power measurement. It can generally be based on a photodiode constructed of semiconductor materials and its related circuits to detect laser power.

[0056] Furthermore, considering that the aforementioned spectrometer module 151 is arranged on the side of the imaging probe 120, its shape will also change with the imaging probe 120 / the excitation light fiber 141 on the imaging probe 120 side. In order to avoid entanglement of the spectrometer module 151 and the power detection module 152 and affect the coupling between the two, the power detection module 152 can be fixed to the electric slip ring rotor end of the hollow electric slip ring 132, and the power detection module 152 can be rotated with the electric slip ring rotor end to avoid torsion between the power detection module 152 and the spectrometer module 151.

[0057] As a result, the power detection module 152 is fixedly connected to the electric slip ring's rotor end and rotates with the optoelectronic composite slip ring 130's rotor end. Based on this fixed connection to the electric slip ring's rotor end, the output end of the power detection module 152 can also be connected to the electric slip ring's rotor end via a metal lead, thereby transmitting power information to the imaging host 110 via the optoelectronic composite slip ring 130.

[0058] In some embodiments, to reduce the coupling accuracy requirements between the light splitting module 151 and the power detection module 152 and avoid coupling failure during rotation, the light splitting module 151 and the power detection module 152 can be coupled via a beam splitting fiber 153. The flexibility of the beam splitting fiber 153 allows for a certain amount of redundancy in the coordination between the light splitting module 151 and the power detection module 152. Specifically, the excitation light detection device 150 further includes a beam splitting fiber 153 connecting the light splitting end of the light splitting module 151 and the input end of the power detection module 152.

[0059] To further illustrate the internal structure of the excitation light detection device, the present application also provides a schematic diagram of the internal structure of the excitation light detection device ( Figure 3 ).

[0060] In some embodiments, to improve the light splitting effect, such as Figure 3 As shown, the aforementioned spectrometer module 151 is configured as a quartz spectrometer, wherein the quartz spectrometer may include a quartz structure 1511 for splitting light and a coupling structure 1512 (such as a coupling lens group) for aligning the output end. The input end of the spectrometer module 151 is coupled to the quartz structure 1511 so that the excitation light reaches the quartz structure. Considering that the expansion coefficient of the quartz structure itself is relatively low, when the excitation light enters the quartz structure 1511 from the core of the excitation light optical fiber, most of the excitation light passes through the quartz structure 1511 and part of the excitation light is split at the quartz structure. The excitation light that passes through the quartz structure 1511 can be output from the output end of the spectrometer module 151 through the coupling structure 1512 (or other coupling structures) and enter the excitation light path / excitation light optical fiber. The excitation light split at the quartz structure can be output from the splitting end to the power detection module 152.

[0061] At the same time, considering that the expansion coefficient of the quartz structure 1511 is low, the original light beam can be evaluated by the light beam split out of it. At the same time, the quartz structure 1511 can reduce the requirements of the splitting optical fiber 153 and conventional optical fiber (such as solid optical fiber) can be used.

[0062] In some embodiments, considering that the excitation light fiber often carries excitation light of multiple wavelengths, the excitation light power of each wavelength can also be detected during power detection to precisely control the excitation light.

[0063] In some embodiments, as Figure 3 As shown, the aforementioned detection of each excitation light can be performed by beam splitting detection during detection, that is, the power detection module 152 can include a beam splitting structure 1521 and a plurality of power sensors 1522. The input end of the beam splitting structure 1521 is coupled to the beam splitting end of the spectrometer module 151, and the plurality of output ends are respectively coupled to the respective power sensors, which are used to split the excitation light split by the spectrometer module 151 based on the wavelength to determine the laser beam corresponding to each excitation light in the plurality of excitation lights. Generally, it can be composed of optical elements (such as beam splitters and filter plates). The power sensor 1522 corresponds to each excitation light in the plurality of excitation lights, and is used to detect the power information of the corresponding laser beam, such as the aforementioned PD. At this time, the beam splitting optical fiber 153 between the spectrometer module 151 and the power detection module 152 can preferably adopt the same optical fiber as the excitation light optical fiber to propagate excitation lights of different wavelengths.

[0064] As an alternative embodiment, the aforementioned detection of each excitation light beam can be performed by splitting the beams into different wavelengths during the beam splitting process. That is, the aforementioned optical splitting module 151 may include multiple optical splitting terminals, wherein the multiple optical splitting terminals are used to split the excitation light beams based on wavelength to determine the laser beam corresponding to each excitation light beam in the multiple excitation light beams. Correspondingly, the power detection module 152 may include multiple power sensors corresponding to each excitation light beam in the multiple excitation light beams, and the power sensors are used to detect the power information of the corresponding laser beam.

[0065] In the aforementioned Figure 2 、 3 In the figure, the spectrometer module 151 can be understood as being arranged on the excitation light fiber on the side of the imaging probe 120. In actual applications, based on its actual setting requirements, the present application also creatively arranges the spectrometer module 151 in the excitation light path of the imaging probe, and makes the beam splitting fiber 153 similar to the fluorescent fiber 142, connected between the imaging probe 120 and the optoelectronic composite slip ring 130.

[0066] To further illustrate the internal structure of the imaging probe 120 provided with the spectrometer module 151 , the present application uses an imaging probe in two-photon fluorescence imaging as an example.

[0067] like Figure 4As shown, the present application also provides a schematic structural diagram of an imaging probe. The imaging probe 120 can be connected to an excitation light fiber 141, a fluorescent fiber 142, and a beam splitting fiber 153 in an optoelectronic composite cable 140 on the imaging probe 120 side. The imaging probe 120 can include a collimator module 121, a scanning galvanometer 122, a lens module 123, and an objective lens module 124. The collimator module 121, the scanning galvanometer 122, the lens module 123, and the objective lens module 124 form an excitation light path with the excitation light fiber 141, and the objective lens module 124 forms a fluorescent light path with the fluorescent fiber 142.

[0068] In the excitation light path, the focus position of the excitation light can be adjusted based on the aforementioned scanning galvanometer 122, focusing on a specific location to determine the fluorescence signal at that location. Other modules are generally presented as optical elements or a collection of them. The specific imaging principles can be found in related technologies of two-photon imaging and are not detailed here.

[0069] To prevent the spectrometer module 151 from affecting the excitation light beam, the spectrometer module 151 can be positioned upstream of the collimator module 121, between the collimator module 121 and the scanning galvanometer 122, or within the lens module 123. This prevents the spectrometer module 151 from affecting the excitation light path. Considering the complexity of the optical components within the excitation light path, the spectrometer module 151 can be configured without a coupling module. Instead, the components within the excitation light path can perform the coupling. This requires only that the split beams be emitted from a location separate from the excitation light fiber 141 and the fluorescent fiber 142.

[0070] like Figure 4 As shown, the splitting module 151 is arranged upstream of the collimating module 121, and is specifically presented as a tilted lens and a reflector arranged upstream of the collimating module 121. The tilted lens can make part of the light beam be split to the side when the excitation light passes through the tilted lens and coupled to the splitting optical fiber 153 by the reflector.

[0071] Exemplary Imaging Control Methods:

[0072] Based on the aforementioned description of the multi-photon imaging device and its excitation light detection device, the imaging control method provided in this application can be executed by the multi-photon imaging device (such as the controller in its imaging host).

[0073] To further describe the imaging control process, the present application also provides an exemplary flow chart of an imaging control method ( Figure 5 ).

[0074] like Figure 5 As shown, process P500 may include the following steps:

[0075] S510 , determining the operating power of the excitation light optical fiber through an excitation light detection device.

[0076] S520 , in response to the relative rotation of the optoelectronic composite slip ring, determining the post-rotation power of the excitation light optical fiber after rotation by an excitation light detection device.

[0077] S530 , adjusting the working parameters of the imaging host based on the working power and the power after rotation, so as to generate a fluorescent image of the imaging object after the excitation light fiber is rotated.

[0078] In the aforementioned S510, the operating power may refer to the power value in the imaging information detected by the excitation light detection device when the imaging device is performing normal imaging before the relative rotation of the optoelectronic composite slip ring. That is, the imaging device can perform normal imaging based on the operating power.

[0079] It should be noted that, considering the actual splitting situation, the operating power is not the actual power of the excitation light during imaging, but rather the power of the split excitation light beam detected by the excitation light detection device. Considering that the two can be linearly / positively correlated through optical design, in practical applications, the operating power can be used as a representative value for the normal operation of the excitation light.

[0080] In some embodiments, when determining S510, taking into account the complexity of the actual working environment, the aforementioned working power may also have certain fluctuations, and an average value over a period of time may also be calculated when determining the working power.

[0081] Similar to the aforementioned working power, in the aforementioned S520 , the power after rotation may refer to a representative value of the excitation light power of the rotated excitation light fiber.

[0082] Based on the positive correlation / linear relationship between the aforementioned power information and the actual excitation light power, when the power after rotation is different from the working power, it can be reflected that the relative rotation of the aforementioned optoelectronic composite slip ring changes the coupling relationship of the light beam, thereby affecting the transmission of the excitation light and possibly causing gain / loss.

[0083] As mentioned above, in actual imaging, a known loss is often expected. Therefore, when there is a difference between the working power and the power after rotation, the above S230 can be executed to adjust the overall loss to a known value of the system, and then imaging is performed based on the known overall loss.

[0084] In some embodiments, the aforementioned adjustment process can be a negative feedback process for the laser output power. That is, the output power of the laser in the imaging host can be adjusted based on the difference between the operating power and the power after rotation, until the power information of the rotated excitation light fiber matches the operating power.

[0085] Furthermore, considering that the losses at different relative angles between the smooth ring stator end and the smooth ring rotor end may be different, when performing compensation, the relative angle between the smooth ring stator end and the smooth ring rotor end can be adjusted to the desired angle corresponding to the working power, and then the power of the aforementioned laser can be adjusted.

[0086] Therefore, based on the aforementioned imaging control method, the imaging parameters can be returned to a known domain so that the influence caused by the rotation of the optoelectronic composite slip ring can be controlled, thereby avoiding its influence on the imaging process.

[0087] In some embodiments, considering that the imaging process provided in the present application often requires plane scanning and / or depth scanning of multiple points, a single imaging task has a certain execution time, and the readjustment of the aforementioned rotation process may affect the execution of the imaging task, timing control can be introduced when rotating to avoid rotation during the imaging process.

[0088] To further illustrate the process, the present application also provides an exemplary flow chart of a rotation control and compensation process ( Figure 6 ).

[0089] like Figure 6 As shown, process P600 may include the following steps:

[0090] S610 : In response to the twisting condition of the excitation light fiber meeting the untwisting condition, the imaging host determines whether the scanning imaging is completed.

[0091] S620: Drive the optoelectronic composite slip ring to rotate based on the torsion of the excitation light fiber.

[0092] S630: Wait for the scanning and imaging to be completed.

[0093] Among them, S610 can be a judgment step, S620 can be a step of branching if yes, and S630 can be a step of branching if no. After S630 is executed, it can jump back to S610 until it meets the corresponding adjustment and executes S620.

[0094] In the aforementioned S610, the detwisting condition may refer to the condition for whether to detwist the cable torsion using an optoelectronic composite slip ring. Considering that detwisting using an optoelectronic composite slip ring in this application has minimal external impact, the aforementioned detwisting adjustment generally requires that the optoelectronic composite slip ring detwist the cable when a full turn of torsion exists. Therefore, based on this detwisting condition, the cable detwisting process is more frequent, ensuring that the cable is in a low-twist / zero-twist state for a long time. If the aforementioned S620 is not executed for an extended period of time, this state can be maintained for a longer period of time without affecting the activity of the imaging subject.

[0095] The aforementioned S610 can generally be implemented based on a timing control signal from the imaging host. That is, the imaging host processor can send timing control signals to various devices. During imaging, the fluorescence signal can be correlated with time based on the timing control signal to render an image. The aforementioned completion of scanning imaging can be understood as the completion of the execution device (such as a scanning galvanometer, an electrically controlled focusing device / a translation stage) executing the corresponding timing control signal (or its corresponding drive signal), while the incomplete scanning imaging can be understood as the incomplete execution of the timing control signal.

[0096] Therefore, based on the judgment process of the aforementioned S610, the aforementioned S620 can only perform rotational detwisting in the gap between two scanning imaging to avoid uncontrollable effects caused by detwisting of the optoelectronic composite slip ring during the imaging process, thereby wasting imaging data.

[0097] In addition, combined with the above discussion on the different coupling conditions at different relative angles, a full circle rotation can be performed when executing the above S620 to ensure that the relative angle between the stator end of the smooth ring and the rotor end of the smooth ring is consistent after each rotation of the central smooth ring, thereby avoiding the impact of its change on power.

[0098] Considering that the optoelectronic composite slip ring will not untwist and rotate during the imaging process, the power adjustment is also mistakenly required to strictly adjust the power back to the power before rotation (i.e., the aforementioned working power). It is only necessary to ensure that it is within the effective range. Its impact on fluorescence can be compensated by the algorithm.

[0099] Specifically, based on the aforementioned P600, after the aforementioned S620, P600 may further include the following steps:

[0100] S640: Determine the power after rotation.

[0101] S650: Determine whether the power after rotation meets the fluorescence imaging requirement.

[0102] S660 , adjusting imaging parameters of the fluorescence image based on the post-rotation power and calibrating the post-rotation power as the working power.

[0103] S670: Adjust the output power of the laser in the imaging host based on the difference between the working power and the power after rotation, until the power information of the rotation of the excitation light fiber is consistent with the working power.

[0104] The aforementioned S650 is a judgment step, and its subsequent step S660 is its yes branch, and S670 is its no branch.

[0105] In the aforementioned S650, the fluorescence imaging requirements can be formed based on experimental testing to determine the power range of the excitation light that can be used for imaging. In addition, compensation for other parameters can also be determined during testing and applied in subsequent steps.

[0106] That is, in S660, after rotation is complete, if the post-rotation power meets the fluorescence imaging requirements, imaging can proceed directly based on the current parameters without adjusting the actual operating parameters of the actuator, thus avoiding other issues. Any impact caused by this can be compensated at the algorithm level. For example, the fluorescence threshold corresponding to the excitation light can be lowered.

[0107] The aforementioned S670 is similar to the aforementioned S530 and will not be described in detail here.

[0108] In some embodiments, during the adjustment / compensation process of the aforementioned P500 and P600, if it is found that the power cannot be adjusted to the working power, it can be determined that an abnormality currently exists and an error is reported to notify the staff to perform manual maintenance.

[0109] During the execution of the aforementioned P500 and P600, the specific power information used can be determined based on the actual hardware configuration and accuracy requirements. For example, the power information can reflect the overall power of the excitation light. For another example, the power information can include the power information of each wavelength beam within the excitation light. However, the feedback and execution processes are similar and are not further described in this application.

[0110] All of the above optional technical solutions can be arbitrarily combined to form optional embodiments of the present application, and will not be described in detail here.

[0111] Those skilled in the art will appreciate that the units and algorithm steps of each example described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, or a combination of computer software and electronic hardware. Whether these functions are performed in hardware or software depends on the specific application and design constraints of the technical solution. Professional and technical personnel can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.

[0112] Those skilled in the art will clearly understand that, for the convenience and brevity of description, the specific working processes of the systems, devices and units described above can refer to the corresponding processes in the aforementioned method embodiments and will not be repeated here.

[0113] In the several embodiments provided in this application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the units is only a logical function division. There may be other division methods in actual implementation, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be through some interfaces, indirect coupling or communication connection of devices or units, which can be electrical, mechanical or other forms.

[0114] The units described as separate components may or may not be physically separate, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed across multiple network units. Some or all of these units may be selected to achieve the purpose of this embodiment according to actual needs.

[0115] It should be noted that, in the description of this application, the terms "first," "second," "third," etc. are used for descriptive purposes only and should not be understood as indicating or implying relative importance. In addition, in the description of this application, unless otherwise specified, "plurality" means two or more.

[0116] It should be noted that "miniaturization" in this application means that the multiphoton microscopy system has little impact on the activities of the living organism to be observed during the observation period. For example, when the multiphoton microscopy system is fixed on the living organism to be observed, the living organism to be observed can still move freely.

[0117] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent replacements, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. An imaging control method for a multi-photon imaging device, characterized in that: The multiphoton imaging device comprises: An imaging host, configured to release excitation light and generate a fluorescence image based on the returned fluorescence signal; An imaging probe, configured to release excitation light to an imaging object to trigger a multiphoton fluorescence effect and collect fluorescence signals generated by the multiphoton fluorescence effect; an optoelectronic composite slip ring, wherein the imaging host is connected to the stator end of the optoelectronic composite slip ring, the imaging probe is connected to the mover end of the optoelectronic composite slip ring, the mover end is coupled to the stator end and driven to rotate relative to the stator end, and the optoelectronic composite slip ring includes a slip ring connected to the excitation light fiber and an electric slip ring connected to at least one metal wire; and an excitation light detection device, one end of which is connected to the smooth ring mover end of the smooth ring, and the other end of which is connected to the electric slip ring mover end of the electric slip ring, for collecting power information of the excitation light in the excitation light optical fiber at the smooth ring mover end; The imaging control method comprises: Determining the operating power of the excitation light optical fiber by the excitation light detection device; In response to the relative rotation of the optoelectronic composite slip ring, determining the post-rotation power of the excitation light optical fiber after rotation by the excitation light detection device; The operating parameters of the imaging host are adjusted based on the operating power and the post-rotation power to generate a fluorescent image of the imaging object after the excitation light fiber is rotated.

2. The imaging control method according to claim 1, wherein: The adjusting the working parameters of the imaging host based on the working power and the post-rotation power includes: The output power of the laser in the imaging host is adjusted based on the difference between the working power and the power after rotation, until the power information of the excitation light fiber after rotation is consistent with the working power.

3. The imaging control method according to claim 1, wherein: The imaging control method comprises: In response to the twisting condition of the excitation light fiber meeting the untwisting condition, the imaging host determines whether scanning imaging is completed; If so, the optoelectronic composite slip ring is driven to rotate based on the torsion of the excitation light fiber.

4. The imaging control method according to claim 3, characterized in that: The adjusting the working parameters of the imaging host based on the working power and the post-rotation power includes: Determining whether the power after rotation meets the fluorescence imaging requirements; If so, adjusting the imaging parameters of the fluorescence image based on the post-rotation power and calibrating the post-rotation power as the working power; If not, the output power of the laser in the imaging host is adjusted based on the difference between the working power and the power after rotation, until the power information of the excitation light fiber after rotation is consistent with the working power.

5. The imaging control method according to claim 1, wherein: The excitation light fiber is configured as a photonic crystal fiber, and the photonic crystal fiber is used to transmit a plurality of excitation lights of different wavelengths within a target wavelength range; The excitation light detection device includes a spectrometer module and a power detection module. The input end of the spectrometer module is connected to the smooth ring mover end, and the output end is connected to the excitation light path of the imaging probe. The spectrometer end is used to split the excitation light from the smooth ring mover end and the excitation light path; the input end of the power detection module is coupled to the spectrometer end of the spectrometer module, and the output end is connected to the electric slip ring mover end, and is used to determine the power information of the split excitation light; The power detection module is fixedly connected to the mover end of the electric slip ring and rotates along with the mover end of the optoelectronic composite slip ring.

6. The imaging control method according to claim 5, characterized in that: The excitation light detection device further includes a beam splitting optical fiber connecting the light splitting end of the light splitting module and the input end of the power detection module.

7. The imaging control method according to claim 6, characterized in that: The optical splitter module is configured as a quartz optical splitter tube, and the beam splitting optical fiber is configured as a solid optical fiber.

8. The imaging control method according to claim 7, characterized in that: The light splitting module is arranged in the excitation light path of the imaging probe, one end of the beam splitting optical fiber is connected to the imaging probe and coupled with the beam splitting end of the light splitting module, and the other end is connected to the power detection module.

9. The imaging control method according to claim 5, characterized in that: The power detection module includes a beam splitting structure and multiple power sensors, the input end of the beam splitting structure is coupled to the beam splitting end of the beam splitting module, and the multiple output ends are respectively coupled to the respective power sensors, and is used to split the excitation light split by the beam splitting module based on wavelength to determine the laser beam corresponding to each excitation light in the multiple excitation lights; the power sensor corresponds to each excitation light in the multiple excitation lights, and is used to detect the power information of the corresponding laser beam; or The spectrometer module includes multiple spectrometer ends, which are used to split the excitation light based on wavelength to determine the laser beam corresponding to each excitation light in the multiple excitation lights. The power detection module includes multiple power sensors corresponding to each excitation light in the multiple excitation lights, and the power sensors are used to detect the power information of the corresponding laser beam.

10. A multiphoton imaging system, characterized in that: The multiphoton imaging system comprises: A multiphoton imaging device corresponding to the imaging control method according to any one of claims 1 to 9; A controller, configured to execute the imaging control method according to any one of claims 1 to 9 for the multiphoton imaging device; and an imaging space for accommodating an imaging object; The imaging probe of the multi-photon imaging device is connected to the imaging object. The optoelectronic composite slip ring of the multi-photon imaging device is arranged at the edge of the imaging space based on the connection direction of the imaging probe and the imaging object. The optoelectronic composite slip ring is used to rotate based on the rotation of the imaging probe relative to the optoelectronic composite slip ring. The excitation light detection device of the multi-photon imaging device is used to detect the power information of the excitation light between the smooth ring rotor end of the optoelectronic composite slip ring and the excitation light path of the imaging probe to adjust the imaging process.

Images