Four-color optical fiber recording system
By using a four-color fiber optic recording system, combined with multiple LED excitation sources and CMOS sensors, the problems of fluorescence signal quenching and low signal-to-noise ratio in existing technologies have been solved, enabling fluorescence signal recording in deeper brain regions and multi-dimensional experimental data acquisition.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-23
- Publication Date
- 2026-03-24
AI Technical Summary
In existing fiber optic recording systems, the excitation sources of violet and blue light cause fluorescence signal quenching, and the signal-to-noise ratio of the yellow light channel is not high, which limits the study of physiological rhythms. Furthermore, the red fluorescent virus is immature and cannot meet the needs of multidimensional characterization of mouse brain activity.
A four-color fiber optic recording system is adopted, which combines a 405nm/470nm/560nm/640nm LED excitation source, a CMOS sensor camera, a data acquisition card, and a microcontroller control circuit board to achieve synchronous recording and analysis of multiple fluorescence signals. The excitation light is coupled to a multimode fiber and the fluorescence signal is reflected to the camera through the optical path components.
The addition of a 640nm red light excitation source improves penetration, enabling the recording of fluorescence signal changes in deeper brain regions. It also has a lower bias value, meeting the needs of multi-dimensional experiments and providing more comprehensive data on fluorescence signal changes in brain regions.
Smart Images
Figure CN224023555U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to fluorescent signal acquisition equipment especially relates to a four color fiber recording system. BACKGROUND
[0002] In recent years, fiber recording system is widely applied in detecting and recording the population calcium signal level of each functional brain area in the process of mouse behavior. The technology detects and records the specific brain area of synchronous mouse behavior activity by drop fluorescent excitation, conduction of multimode optical fiber, and combination of weak light signal detection and specific fluorescent marking technology. The excitation light emitted by the light source is reflected and focused in the multimode optical fiber by the dichroic mirror, the optical fiber transmits the excitation light to the specific brain area, excites the neuron specifically marked by the calcium indicator, and the neuron activity is shown in the form of fluorescence. The neuron activity or neurotransmitter level is positively correlated with the fluorescence signal intensity, and as the neuron activity or neurotransmitter level increases, the fluorescence signal also increases, and vice versa. The transformed fluorescence signal is transmitted back to the detector through the same optical fiber, and is converted into an image signal that can be collected through a photoelectric conversion device, thereby reflecting the function of the related neural nucleus when the specific behavior occurs.
[0003] At present, the excitation light of the common fiber recording system is generally 3 kinds of light sources, which are 405nm purple light, 470nm blue light and 560nm yellow light. Among them, the purple light and the blue light will excite the fluorescence of the multimode optical fiber and cause the fluorescence signal in the mouse brain to be obviously quenched during the recording process, which limits the development of some long-term research projects such as the study of physiological rhythm. Secondly, the virus of red fluorescence is not mature enough, so the data signal-to-noise ratio collected by the 560nm yellow light channel is not high. Finally, with the wide application of the fiber recording system in China, the research demand is also expanding, and it is necessary to characterize the activity of different regions in the mouse brain from more dimensions.
[0004] Therefore, we propose a four-color fiber recording system to solve the above problems. UTILITY MODEL CONTENT
[0005] The technical problem to be solved by the utility model is to overcome the defects in the prior art. The utility model provides a four-color fiber recording system. Through the combination of the acquisition card, the (405nm / 470nm / 560nnm / 640nm) four kinds of LED excitation light source combination, the integrated single-chip microcomputer control circuit board, the optical path assembly, the camera, the case and the acquisition and analysis software, the synchronous recording and analysis of multiple fluorescence signals are realized.
[0006] To solve the above technical problems, the utility model adopts the technical scheme of a four-color fiber recording system, which comprises:
[0007] The excitation light source comprises at least 4 kinds of LED lasers.
[0008] a camera, which is internally provided with a CMOS sensor to acquire a time sequence diagram corresponding to a fluorescent signal in a specific brain region of an experimental animal;
[0009] a collection card, which communicates with an external system terminal, and is used to realize collection of a TTL signal output by the system and a TTL signal input by an external system;
[0010] a control circuit, which is internally integrated with a single-chip microcomputer, and is used to realize control of different light-emitting sequences of a plurality of LED lasers and synchronous collection of the camera and different wavelength excitation lights;
[0011] an optical path assembly, which is used to couple the plurality of LED lasers into a same multimode optical fiber and transmit them to a specific brain region of an experimental animal, and reflect returned fluorescent signals to the camera.
[0012] Further, the excitation light source is four LED lasers, wherein central wavelengths of three LED lasers are 405 nm, 470 nm and 560 nm respectively, and a central wavelength range of another LED laser is 630-710 nm.
[0013] Further, the optical path assembly comprises optical filters, a mirror and a dichroic mirror, each of the LED lasers in the excitation light source is filtered through an independent optical filter, then is refracted by the dichroic mirror and input into the specific brain region of the experimental animal through an optical fiber, and finally is reflected by the mirror and filtered according to wavelengths to be introduced into the CMOS sensor.
[0014] Further, the camera comprises two cameras, one of which collects LED lasers with central wavelengths of 405 nm and 470 nm, and the other of which collects LED lasers with a central wavelength of 560 nm and the remaining one.
[0015] Further, the single-chip microcomputer is of an MK60DN512 type, and corresponding pins thereof are electrically connected with the excitation light source, the camera and the collection card.
[0016] Further, the collection card is of an NI6001 collection card of the National Instruments, USA.
[0017] Further, the control circuit can be integrated in a circuit board, and the circuit board, the excitation light source, the camera, the collection card and the optical path assembly are uniformly installed and positioned by an external case.
[0018] Compared with the prior art, the beneficial effects of the utility model include: through increasing 640nm red light as excitation light source in optical fiber system, using the characteristics of stronger penetration of excitation light of this wavelength, can record fluorescence signal change of deeper brain area, the bias value is also lower when collecting signal, can react fluorescence signal change in brain area from more dimensions, can satisfy more experimental design needs. BRIEF DESCRIPTION OF DRAWINGS
[0019] The disclosure of the utility model will be explained with reference to the drawings. It should be appreciated that the drawings are only for illustrative purposes and are not intended to limit the scope of protection of the utility model. In the drawings, the same reference signs are used to refer to the same parts. Among them:
[0020] Figure 1 The system overall structure schematic diagram according to an embodiment of the utility model is schematically shown;
[0021] Figure 2 The single-chip microcomputer circuit connection principle diagram according to an embodiment of the utility model is schematically shown;
[0022] Figure 3 The excitation light source circuit principle diagram according to an embodiment of the utility model is schematically shown;
[0023] Figure 4a And Figure 4b The control circuit principle diagram of the center wavelength 405nm LED laser according to an embodiment of the utility model is schematically shown.
[0024] Reference signs in the drawings: 1, first camera; 2, second camera; 3, reflector; 4, optical filter; 5, dichroic mirror. DETAILED DESCRIPTION
[0025] It is easy to understand that according to the technical scheme of the utility model, a person skilled in the art can propose a plurality of structure modes and implementation modes that can be replaced with each other without changing the essential spirit of the utility model. Therefore, the following specific embodiments and drawings are only exemplary description of the technical scheme of the utility model, and should not be regarded as the whole or regarded as the limitation or restriction of the technical scheme of the utility model.
[0026] According to an embodiment of the utility model Figure 1 , Figure 2 , Figure 3 , Figure 4a and Figure 4b are shown.
[0027] As Figure 1As shown, in this embodiment, a four-color fiber optic recording system uses two cameras to simultaneously record four excitation fluorescence signals (405nm / 470nm / 560nm / 640nm). The system uses a microcontroller to synchronously control the light source and cameras, and employs time-division multiplexing technology to distinguish fluorescence signals from different channels. Four types of LED lasers emitted from the excitation source are coupled into a single beam through multiple dichroic mirrors, and finally focused into a multimode fiber after reflection by the dichroic mirrors. The fiber transmits the excitation light to a specific brain region, stimulating neurons specifically labeled with calcium indicators, and the fluorescence signal changes generated by neuronal activity are reflected back to the system's detector through the same multimode fiber.
[0028] The fluorescence signals excited by excitation light with wavelengths of 405nm and 470nm are green wavelengths and are acquired by the system's first camera. The synchronization of the switching timing of the first camera with the 405nm and 470nm excitation light is controlled by the system's microcontroller. The fluorescence signals excited by excitation light with wavelengths of 560nm and 640nm are red and far-red wavelengths, respectively and are acquired by the system's second camera. The synchronization of the switching timing of the second camera with the 560nm and 640nm excitation light is also controlled by the system's microcontroller. The first and second cameras respectively set the odd-numbered frames and even-numbered frames acquired as channel data for the two fluorescence signals. The switching timing of the above four light sources does not overlap in time.
[0029] like Figure 2 As shown, in this embodiment, the specific microcontroller signal model is MK60DN512. Pins 20 and 26 are connected to the first and second cameras respectively, pins 21, 22, 23, and 24 are connected to the four types of LED lasers respectively, and pin 25 is connected to the acquisition card. The connection methods of the four types of LED lasers to each connector are as follows: Figure 3 As shown.
[0030] Similarly, such as Figure 4a and Figure 4b As shown, for each type of LED laser control circuit, taking purple as an example, the U7 chip, together with an external power supply, provides a constant current to the LED laser. Then, the LED pp- is connected to the corresponding relay's interface 3. The corresponding LED driver line is controlled to turn on and off through the relay, thus realizing the LED's on and off state.
[0031] It can be seen that the optical path assembly described above can couple four excitation lights into the same multi-mode optical fiber and transmit them into the mouse brain, and reflect the returned fluorescent signals to two cameras; the cameras are the fluorescent signal detectors of the optical fiber recording system, the multi-mode optical fiber is imaged through the optical path assembly, and the gray value of the imaging area of the multi-mode optical fiber is processed through the acquisition software to finally realize the acquisition function of the fluorescent signal; the acquisition software can communicate with the acquisition card / circuit board of the integrated single-chip microcomputer / camera to realize the switching control and brightness adjustment of the four excitation lights in the system, and the final display and data saving functions of the fluorescent signal; the analysis software can normalize the fluorescent signal saved by the acquisition software, draw the fluorescent change curve graph and heat map, and further statistically analyze the normalized data and analyze and process multiple groups of data.
[0032] Specifically, the frame data returned by the camera is counted and saved by the external acquisition software (such as QAXK-FPS-OST-ACQUISITION-SOFTWARE) for the gray value of the imaging area of the optical fiber, thereby realizing the acquisition of the fluorescent signal. The analysis software can normalize the fluorescent signal data obtained by the acquisition and draw a curve graph and a heat map. At the same time, the system is internally provided with an acquisition card, which can realize the synchronous TTL signal output of the system and the synchronous TTL signal output of other systems, thereby realizing the synchronous analysis of the fluorescent signal.
[0033] The system realizes the synchronous recording and analysis of multiple fluorescent signals through the combination of the acquisition card, the combination of the four LED excitation light sources (405nm / 470nm / 560nnm / 640nm), the control circuit board of the integrated single-chip microcomputer, the optical path assembly, the camera, the case and the acquisition and analysis software. The light source can be customized according to the customer's demand to realize the acquisition of different fluorescent signals.
[0034] The synchronous timing of the light source and the camera can be customized according to the special needs of the user, and the fluorescent signals corresponding to the three excitation lights can be collected by the same camera, the fluorescent signal of one excitation light can be collected alone, and the corresponding timing of the excitation light and the camera can be freely arranged and combined. The four-color optical fiber recording system can be widely used in various fields, such as biomedicine, life science, etc. The whole system can realize the synchronous recording of multiple fluorescent signals through time division multiplexing, and more dimensions can be used to show the activity level of different brain areas; the system can realize the synchronous acquisition of the fluorescent signals of the same site and the light regulation stimulation.
[0035] The technical scope of the utility model is not limited to the content in the above description, and those skilled in the art can make various deformations and modifications to the above embodiment without departing from the technical thought of the utility model, and these deformations and modifications should all belong to the protection scope of the utility model.
Claims
1. A four-color optical fiber recording system characterized by, It comprises: An excitation light source, which is four kinds of LED lasers, three of which have central wavelengths of 405 nm / 470 nm / 560 nm, and the other has a central wavelength range of 630-710 nm adjustable; A camera with a built-in CMOS sensor to obtain the time sequence diagram corresponding to the fluorescence signal in the specific brain area of the experimental animal; An acquisition card for communication with external system terminals to realize the output of system synchronization TTL signals and the acquisition of external input system TTL signals; A control circuit with a built-in single-chip microcomputer to realize the control of different light-emitting sequences of multiple LED lasers and the synchronous acquisition of the camera and different wavelength excitation light; An optical path assembly to couple multiple LED lasers into the same multimode optical fiber and transmit them to the specific brain area of the experimental animal, while reflecting the returned fluorescence signal to the camera.
2. A four-color optical fiber recording system according to claim 1, characterized in that: The optical path assembly includes optical filters, mirrors, and dichroic mirrors. Each LED laser in the excitation light source is filtered by an independent optical filter, then refracted by the dichroic mirror and input into the specific brain area of the experimental animal through the optical fiber, and finally reflected by the mirror while the reflected fluorescence signal is filtered by wavelength and introduced into the CMOS sensor.
3. A four-color optical fiber recording system according to claim 1, characterized in that: The camera includes two cameras, one of which corresponds to the fluorescence collection of LED lasers with central wavelengths of 405 nm and 470 nm, and the other of which corresponds to the fluorescence collection of LED excitation light with a central wavelength of 560 nm and the remaining one.
4. The four-color optical fiber recording system of claim 1, wherein: The model of the single-chip microcomputer is MK60DN512, and its corresponding pins are electrically connected to the excitation light source, camera, and acquisition card.
5. A four-color optical fiber recording system according to claim 4, characterized in that: The model of the acquisition card is the NI6001 acquisition card of the National Instruments of the United States.
6. A four-color optical fiber recording system according to claim 4, characterized in that: The control circuit can be integrated on a circuit board, and the circuit board, excitation light source, camera, acquisition card, and optical path assembly are uniformly installed and positioned by an external case.