Optical fiber recording device

Abstract

The optical fiber recording device provided by the embodiment of the utility model comprises an excitation light module which emits an excitation light signal and irradiates a preset position so as to excite a fluorescence signal; the photoelectric detection module receives a fluorescence signal; the optical assembly comprises an optical fiber, and the optical fiber guides an excitation light signal to a preset position and guides a fluorescence signal to the photoelectric detection module; the acquisition and analysis module is respectively connected with the excitation light module and the photoelectric detection module, acquires the excitation light signal and the fluorescence signal, and performs synchronization and analysis to obtain fluorescence lifetime information in the fluorescence signal. According to the embodiment, the excitation light signal of the excitation light module is guided to the preset position through the optical fiber to excite the fluorescence signal, the fluorescence signal is guided to the photoelectric detection module through the optical fiber, and the excitation light signal and the fluorescence signal are collected and synchronized and analyzed to obtain fluorescence lifetime information in the fluorescence signal. The method is suitable for long-term fluorescence signal analysis, and is also suitable for observing and recording the neural activity of the brain region of a small animal which moves freely.

Description

Technical Field

[0001] This invention relates to the field of fluorescence signal detection technology in living organisms, and more particularly to an optical fiber recording device. Background Technology

[0002] Existing fiber optic recording devices acquire fluorescence signals of a single wavelength or different wavelengths and detect the fluorescence intensity to observe and record neural activity in animal brain regions. However, fluorescence intensity is affected by factors such as probe concentration, excitation light intensity, and photobleaching effect. As the detection time increases, the fluorescence intensity changes significantly, making it unsuitable for long-term, stable fluorescence signal analysis.

[0003] Fluorescence lifetime in fluorescence signals is unaffected by factors such as probe concentration, excitation light intensity, and photobleaching effect, making it suitable for long-term, stable fluorescence signal analysis. Furthermore, fluorescence lifetime offers advantages such as high specificity, high sensitivity, and quantitative measurement in monitoring microenvironmental changes and reflecting intermolecular interactions. Utility Model Content

[0004] This invention provides an optical fiber recording device, which aims to solve the problem that existing optical fiber recording devices are not suitable for long-term stable fluorescence signal analysis.

[0005] In a first aspect, an optical fiber recording device is provided, comprising:

[0006] The excitation light module (1) is used to emit an excitation light signal of at least one wavelength and irradiate a preset position to excite a fluorescence signal of at least one wavelength;

[0007] A photoelectric detection module (2) is used to receive fluorescence signals of at least one wavelength;

[0008] Optical component (3), which includes optical fiber (31), which guides an excitation light signal of at least one wavelength to a preset position and guides a fluorescence signal of at least one wavelength to a photodetector module (2).

[0009] The acquisition and analysis module (4) is connected to the excitation light module (1) and the photodetector module (2) respectively. It is used to acquire excitation light signals of at least one wavelength and fluorescence signals of at least one wavelength, and to synchronize and analyze them to obtain at least one fluorescence lifetime information in the fluorescence signals of at least one wavelength.

[0010] This invention guides the excitation light signal from the excitation light module to a preset position via an optical fiber to excite the fluorescence signal. The fluorescence signal is then guided to the photodetector module via the optical fiber to collect the excitation light signal and the fluorescence signal. The two signals are synchronized and analyzed to obtain fluorescence lifetime information from the fluorescence signal. This invention is suitable for long-term fluorescence signal analysis. Furthermore, due to the small size and flexibility of the optical fiber, it is also suitable for observing and recording the neural activity of brain regions in freely moving small animals. Attached Figure Description

[0011] The above and / or additional aspects and advantages of this utility model will become apparent and readily understood from the following description of the embodiments taken in conjunction with the accompanying drawings, in which:

[0012] Figure 1 This is a schematic diagram of the optical fiber recording device provided in Embodiment 1 of this utility model;

[0013] Figure 2 This is a schematic diagram of the optical fiber recording device provided in Embodiment 2 of this utility model;

[0014] Figure 3 This is another structural schematic diagram of the optical fiber recording device provided in Embodiment 2 of this utility model;

[0015] Figure 4 This is a schematic diagram of the end face of the multimode fiber bundle provided in Embodiment 2 of this utility model;

[0016] Figure 5 This is a schematic diagram of the optical fiber recording device provided in Embodiment 3 of this utility model;

[0017] Figure 6 This is another structural schematic diagram of the optical fiber recording device provided in Embodiment 3 of this utility model. Detailed Implementation

[0018] The embodiments of this utility model are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar modules or modules having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this utility model, and should not be construed as limiting this utility model. Rather, the embodiments of this utility model include all variations, modifications, and equivalents falling within the spirit and scope of the appended claims.

[0019] This invention guides the excitation light signal from the excitation light module to a preset position via an optical fiber to excite the fluorescence signal. The fluorescence signal is then guided to the photodetector module via the optical fiber to collect the excitation light signal and the fluorescence signal. The two signals are synchronized and analyzed to obtain fluorescence lifetime information from the fluorescence signal. This invention is suitable for long-term fluorescence signal analysis. Furthermore, due to the small size and flexibility of the optical fiber, it is also suitable for observing and recording the neural activity of brain regions in freely moving small animals.

[0020] Figure 1 This is a schematic diagram of the optical fiber recording device provided in Embodiment 1 of this utility model. Figure 1 As shown, the fiber optic recording device includes an excitation light module 1, a photoelectric detection module 2, an optical component 3, and an acquisition and analysis module 4.

[0021] The excitation light module 1 includes at least one modulated excitation light source and / or at least one high-repetition-rate ultrashort pulse excitation light source, emitting an excitation light signal of at least one wavelength to irradiate a preset location of a specific neuron in the experimental animal to excite a fluorescence signal of at least one wavelength. The excitation light source is a picosecond laser, femtosecond laser, etc., with a pulse width of less than 600 picoseconds and a repetition frequency greater than 1 MHz and not less than 40 MHz. The wavelength of the excitation light signal is different from the wavelength of the fluorescence signal generated by it.

[0022] The optical component 3 includes an optical fiber 31, which serves as the detection end. The optical fiber 31 guides an excitation light signal of at least one wavelength emitted by the excitation light module 1 to a preset position, and guides a fluorescence signal of at least one wavelength emitted at the preset position to the photodetector module 2. The optical fiber 31 is a low-fluorescence optical fiber, meaning its autofluorescence level is low, reducing interference with the fluorescence signal and improving the accuracy of fluorescence lifetime observation. Preferably, its core diameter is 5-1000 micrometers, such as 50 micrometers.

[0023] The photodetector module 2 receives the fluorescence signal of at least one wavelength and converts the fluorescence signal into an electrical signal. It includes at least one photomultiplier tube, single photon avalanche diode (SPAD), hybrid photon detector (HPD), other array photon detectors, or array photodetectors with picosecond or femtosecond capabilities.

[0024] The acquisition and analysis module 4 is connected to the excitation light module 1 and the photodetector module 2 respectively, and acquires the excitation light signal and the fluorescence signal of the at least one wavelength, and performs synchronization and analysis to obtain at least one fluorescence lifetime information in the fluorescence signal of the at least one wavelength.

[0025] In this embodiment of the invention, the acquisition and analysis module 4 acquires and synchronizes the electrical signals of the excitation light signal and the fluorescence signal, such as aligning the timing of the pulses of each excitation light signal and aligning the timing of the excitation light signal with the fluorescence signal generated by it. Then, it processes, analyzes, and displays the signals based on the relationship between these two signals to observe neuronal activity. The acquisition and analysis module 4 processes the excitation light signal and fluorescence signal using either frequency domain methods or time domain methods. The frequency domain method involves the acquisition and analysis module 4 analyzing the frequency domain information of at least one wavelength of excitation light signal and at least one wavelength of fluorescence signal to obtain at least one fluorescence lifetime information from the fluorescence signal of at least one wavelength. For example, a modulated continuous excitation light signal is used to irradiate a preset position, and the fluorescence lifetime information is calculated and analyzed by detecting changes in the amplitude and phase of the fluorescence signal. The time domain method involves the acquisition and analysis module 4 analyzing the time domain information of at least one wavelength of excitation light signal and at least one wavelength of fluorescence signal to obtain at least one fluorescence lifetime information from the fluorescence signal of at least one wavelength. For example, a preset position can be irradiated with a high repetition frequency ultrashort pulse excitation light signal, and a high-precision time marking device, such as a time-correlated single-photon counter, can be used to record the output time of the excitation light signal in the excitation light module 1, the corresponding reception time and / or intensity of the fluorescence signal in the photodetector module 2, the number of photons, etc., and construct a fluorescence decay curve to analyze and obtain fluorescence lifetime information.

[0026] Furthermore, the acquisition and analysis module 4 can also be used to analyze at least one fluorescence intensity information from fluorescence signals of at least one wavelength, and combine at least one fluorescence intensity information and at least one fluorescence lifetime information to analyze fluorescence signals of at least one wavelength. The fiber optic recording device of this embodiment has the ability to detect both fluorescence intensity information and fluorescence lifetime information. By comparing and analyzing these two information, more neuronal activity can be observed, which is beneficial for more scientific discoveries.

[0027] The acquisition and analysis module 4 can be a single device that performs acquisition, analysis, data storage, and display functions; or it can be divided into different units or devices, with one device performing the acquisition and analysis functions and another device performing the data storage and display functions, presenting the analyzed fluorescence intensity information, fluorescence lifetime information, etc. to the user.

[0028] Figure 2 This is a schematic diagram of the optical fiber recording device provided in Embodiment 2 of this utility model. Figure 2 As shown, optical component 3 also includes a coupling mirror 32 and a first dichroic mirror 33. The coupling mirror 32 can be composed of an aspherical lens or a spherical lens, or it can be a high numerical aperture microscope objective.

[0029] Excitation light signals of at least one wavelength emitted by excitation light module 1 are reflected by first dichroic mirror 33 and enter coupling mirror 32. Coupler 32 couples the excitation light signals of at least one wavelength into optical fiber 31 to guide them to a preset position, thereby exciting a fluorescence signal of at least one wavelength. This fluorescence signal of at least one wavelength is guided through coupling mirror 32 by optical fiber 31 and transmitted through first dichroic mirror 33 before being received by photodetector module 2. In another embodiment of this invention, the positions of excitation light module 1 and photodetector module 2 are interchanged. The excitation light signal is transmitted through first dichroic mirror 33 and enters coupling mirror 32, while the fluorescence signal is reflected by first dichroic mirror 33 and received by photodetector module 2. Acquisition and analysis module 4 is connected to excitation light module 1 and photodetector module 2 respectively. In this embodiment of the invention, optical fiber 31 can be a single-mode optical fiber, and the excitation light signal and the fluorescence signal generated by it pass through the same optical fiber in opposite directions, allowing for a smaller area at the irradiated preset position. To reduce the reflected light when the excitation light signal and fluorescence signal enter the optical fiber 31, the end face of the optical fiber 31 is treated to reduce reflection, such as by making the end face inclined or by coating it with an anti-reflection film.

[0030] Furthermore, Figure 3 This is another structural schematic diagram of the optical fiber recording device provided in Embodiment 2 of this utility model. (See diagram below.) Figure 3As shown, the excitation light module 1 includes a first excitation light source 11, a second excitation light source 12, and a third excitation light source 13, the laser power output by each of the three can be independently adjusted. The first excitation light source 11, the second excitation light source 12, and the third excitation light source 13 emit excitation light signals of different wavelengths in collimated form. Optionally, each of the three has its corresponding filter. The excitation light signal emitted by the third excitation light source 13 is transmitted through a dichroic mirror corresponding to the second excitation light source 12, and converges with the excitation light signal emitted by the second excitation light source 12 reflected by the dichroic mirror to form a coaxial, two-color beam; the excitation light signal emitted by the first excitation light source 11 is transmitted through a corresponding dichroic mirror, and converges with the aforementioned two-color beam reflected by the dichroic mirror to form a coaxial, collimated three-color beam, which reaches the first dichroic mirror 33, and then illuminates a preset position through a coupling mirror 32 and an optical fiber 31, exciting a first fluorescence signal, a second fluorescence signal, and a third fluorescence signal. The wavelengths of the first fluorescence signal, the second fluorescence signal, and the third fluorescence signal do not overlap. The photoelectric detection module 2 includes a first photodetector 21, a second photodetector 22, and a third photodetector 23. Optionally, each of the three has its corresponding filter. The first photodetector 21, the second photodetector 22, and the third photodetector 23 respectively receive fluorescence signals of different wavelengths. After irradiating a preset position, one wavelength of the multi-wavelength fluorescence signal excited is reflected by the dichroic mirror corresponding to the first photodetector 21 and received by the first photodetector 21, while other wavelengths of fluorescence signals are transmitted through the dichroic mirror. Then, among the other wavelengths of fluorescence signals, one wavelength of fluorescence signal is transmitted through the dichroic mirror corresponding to the second photodetector 22 and received by the third photodetector 23, while another fluorescence signal is reflected by the dichroic mirror corresponding to the second photodetector 22 and received by the second photodetector 22.

[0031] Depending on experimental requirements, the excitation light module 1 may include more excitation light sources. Each excitation light source is converged into a beam to illuminate a preset location, exciting fluorescence signals of different wavelengths. Existing technologies can be used to converge the excitation light sources into a single beam and separate the fluorescence signals, which will not be elaborated upon here.

[0032] In order to achieve multi-channel fiber optic recording, as another embodiment of this utility model, fiber 31 is a multimode fiber bundle composed of multiple multimode fibers with a numerical aperture greater than 0.22, and each multimode fiber is a channel. Figure 4 This is a schematic diagram of the end face of the multimode fiber bundle provided in Embodiment 2 of this utility model. Figure 4 As shown, one end of multiple multimode optical fibers is bundled together to form a bundled end face, which is a densely packed multi-branched end face located at the focal plane of the coupling mirror 32. The other ends of the multiple multimode optical fibers are dispersed, and the multimode optical fibers of different branches can irradiate different neurons of the same experimental animal or neurons of different experimental animals.

[0033] Excitation light signals of at least one wavelength are converged into a beam, transmitted or reflected by the first dichroic mirror 33, and then enter the coupling mirror 32. The coupling mirror 32 couples the excitation light signals of at least one wavelength, and the diameter of the flat-top spot of each excitation light signal is not less than the diameter of the bundled end face, so that it enters each multimode fiber uniformly at the bundled end face of the multimode fiber bundle, guiding it to different preset positions to excite fluorescence signals of at least one wavelength. This allows each channel to be experimented independently under the same conditions (such as the same excitation time, the same excitation intensity, etc.). The fluorescence signals of multiple excited channels are guided through the multimode fiber bundle through the coupling mirror 32, and after being reflected or transmitted by the first dichroic mirror 33, they are received by different areas of the photodetector module 2. The photodetector module 2 may include one photodetector or multiple photodetectors. Fluorescence signals of different channels are received by different photodetectors or by different areas or designated pixels of the same photodetector. Fluorescence signals of the same channel are received by the same area of ​​the same photodetector in a time-division manner or by different areas of the same photodetector simultaneously. The photoelectric detection module 2 converts the fluorescence signal into an electrical signal. The acquisition and analysis module 4 obtains the electrical signal of each channel according to the mapping relationship between the photoelectric detectors corresponding to each channel and the specified pixels, and performs independent or comprehensive analysis on each channel.

[0034] Figure 5 This is a schematic diagram of the optical fiber recording device provided in Embodiment 3 of this utility model. Figure 5 As shown, optical fiber 31 includes a first optical fiber 311 and a second optical fiber 312. The first optical fiber 311 guides the excitation light signal of at least one wavelength emitted by the excitation light module 1 to a preset position, and the second optical fiber 312 guides the fluorescence signal of at least one wavelength to the photodetector module 2. The excitation light module 1 and the photodetector module 2 can be designed as in Embodiment 2, and will not be described again here. In this embodiment of the present invention, optical fiber 31 can be a single-mode optical fiber, and the excitation light signal and the fluorescence signal generated by it pass through different optical fibers, reducing the interference between the excitation light signal and the fluorescence signal.

[0035] Furthermore, Figure 6 This is another structural schematic diagram of the optical fiber recording device provided in Embodiment 3 of this utility model. (See diagram below.) Figure 6As shown, the optical component 3 also includes an adapter 34, a first pin 35, and a second pin 36. The first optical fiber 311 is connected to the first pin 35 via the adapter 34, and the second optical fiber 312 is connected to the second pin 36 via the adapter 34, illuminating a preset position. When the experiment reaches a certain stage, the first optical fiber 311 and the second optical fiber 312 can be detached, and the adapter 34, the first pin 35, and the second pin 36, or vice versa, can be reattached to the experimental animal. In the next stage of the experiment, the first optical fiber 311, the second optical fiber 312, and the adapter 34 can be reconnected without requiring further pre-experimental treatment of the experimental animal, thus reducing harm to the animal. The adapter 34, the first pin 35, and the second pin 36 are small in size and lightweight, and will not affect the daily activities of the experimental animal.

[0036] This invention guides the excitation light signal from the excitation light module to a preset position via an optical fiber to excite the fluorescence signal. The fluorescence signal is then guided to the photodetector module via the optical fiber to collect the excitation light signal and the fluorescence signal. The two signals are synchronized and analyzed to obtain fluorescence lifetime information from the fluorescence signal. This invention is suitable for long-term fluorescence signal analysis. Furthermore, due to the small size and flexibility of the optical fiber, it is also suitable for observing and recording the neural activity of brain regions in freely moving small animals.

[0037] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. An optical fiber recording device, characterized in that, include: The excitation light module (1) is used to emit an excitation light signal of at least one wavelength and irradiate a preset position to excite a fluorescence signal of at least one wavelength; A photoelectric detection module (2) is used to receive fluorescence signals of at least one wavelength; Optical component (3), which includes optical fiber (31), the optical fiber (31) guides the excitation light signal of the at least one wavelength to a preset position and guides the fluorescence signal of the at least one wavelength to the photodetector module (2). The acquisition and analysis module (4) is connected to the excitation light module (1) and the photoelectric detection module (2) respectively, and is used to acquire the excitation light signal of at least one wavelength and the fluorescence signal of at least one wavelength, and to synchronize and analyze them to obtain at least one fluorescence lifetime information in the fluorescence signal of at least one wavelength.

2. The optical fiber recording device according to claim 1, characterized in that, The optical component (3) also includes a coupling mirror (32) and a first dichroic mirror (33); The excitation light signal of at least one wavelength is transmitted or reflected by the first dichroic mirror (33) and then enters the coupling mirror (32). The coupling mirror (32) couples the excitation light signal of at least one wavelength into the optical fiber (31) to guide it to a preset position and excite the fluorescence signal of at least one wavelength. The fluorescence signal of at least one wavelength is guided through the optical fiber (31) through the coupling mirror (32), and is received by the photoelectric detection module (2) after being reflected or transmitted by the first dichroic mirror (33).

3. The optical fiber recording device according to claim 2, characterized in that, The excitation light module (1) includes a first excitation light source (11), a second excitation light source (12) and a third excitation light source (13), and the photoelectric detection module (2) includes a first photodetector (21), a second photodetector (22) and a third photodetector (23).

4. The optical fiber recording device according to claim 2, characterized in that, The optical fiber (31) is a multimode fiber bundle; The excitation light signal of at least one wavelength is transmitted or reflected by the first dichroic mirror (33) and then enters the coupling mirror (32). The coupling mirror (32) couples the excitation light signal of at least one wavelength into the multimode fiber bundle to guide it to different preset positions and excite the fluorescence signal of at least one wavelength. The fluorescence signal of at least one wavelength is guided through the multimode fiber bundle through the coupling mirror (32), and after being reflected or transmitted by the first dichroic mirror (33), it is received by different regions of the photoelectric detection module (2).

5. The optical fiber recording device according to claim 1, characterized in that, The optical fiber (31) includes a first optical fiber (311) and a second optical fiber (312); The first optical fiber (311) guides the excitation light signal of at least one wavelength to a preset position; The second optical fiber (312) guides the fluorescence signal of the at least one wavelength to the photodetector module (2).

6. The optical fiber recording device according to claim 5, characterized in that, The optical component (3) also includes an adapter (34), a first pin (35), and a second pin (36); The first optical fiber (311) is connected to the first ferrule (35) via the adapter (34); The second optical fiber (312) is connected to the second ferrule (36) via the adapter (34).

7. The optical fiber recording apparatus according to any one of claims 1-6, characterized in that, The excitation light module (1) includes at least one modulated excitation light source and / or at least one high repetition frequency ultrashort pulse excitation light source; The photoelectric detection module (2) includes at least one photomultiplier tube, single-photon avalanche diode or hybrid photodetector.

8. The optical fiber recording apparatus according to any one of claims 1-6, characterized in that, The acquisition and analysis module (4) is used for: Based on the frequency domain information of the excitation light signal of the at least one wavelength and the fluorescence signal of the at least one wavelength, at least one fluorescence lifetime information of the fluorescence signal of the at least one wavelength is obtained by analysis; or Based on the time-domain information of the excitation light signal of the at least one wavelength and the fluorescence signal of the at least one wavelength, at least one fluorescence lifetime information of the fluorescence signal of the at least one wavelength is obtained by analysis.

9. The optical fiber recording apparatus according to any one of claims 1-6, characterized in that, The acquisition and analysis module (4) is also used for: Analysis yields at least one fluorescence intensity information from the fluorescence signals of at least one wavelength; The fluorescence signal at the at least one wavelength is analyzed by combining the at least one fluorescence intensity information and the at least one fluorescence lifetime information.

10. The optical fiber recording apparatus according to any one of claims 1-6, characterized in that, The core diameter of the optical fiber 31 is 5-1000 micrometers.

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