Two-color ultra-miniature microscopic imaging system

Abstract

The utility model provides a double-color ultra-miniature microscopic imaging system, which solves the problem that the accuracy and effectiveness of data cannot be judged by the existing single-color fluorescence recording, and adopts the main scheme that the double-color ultra-miniature microscopic imaging system comprises a video acquisition card, a double-color ultra-miniature microscopic imaging mirror body, a multi-core optical fiber bundle, an LED (light-emitting diode) driving circuit, a digital signal acquisition card and a switching power supply, the video acquisition card is electrically connected with the LED driving circuit and can be communicated with computer acquisition software to distinguish odd frames and even frames by taking image frames as units and synchronously control corresponding light source signals, and the double-color ultra-miniature microscopic imaging mirror body is used for emitting two excitation lights and collecting and imaging fluorescence models. The multi-core optical fiber bundle is used for coupling two kinds of exciting light into a light source and connecting the light source to the two-color ultra-miniature microscopic imaging mirror body, and the LED driving circuit is used for receiving a control signal of the video acquisition card so as to realize synchronization of on-off of the exciting light and image frames.

Description

TECHNICAL FIELD

[0001] The utility model relates to microscope technical field especially relates to a double -color ultramicro microscope imaging system. BACKGROUND

[0002] The ultramicro microscope imaging system is combined calcium ion fluorescence indicator, and through implanting GRIN (gradient refractive index) lens in the brain area of animal, can realize deep brain imaging, and record the calcium signal of a group of neurons at single cell level.The system has the advantages of small volume, light weight, does not affect the free movement and behavior experiment of mouse.Because of the two index requirements of small volume and light weight, the commonly used ultramicro microscope imaging system at present mostly carries out excitation in monochrome, and only records green fluorescence;Further, the research range of monochrome excitation light is smaller, which limits the experimental design of user, the motion noise of ultramicro microscope imaging system brought by free movement of animal is larger, and monochrome fluorescence recording cannot judge the accuracy and effectiveness of data, therefore, it is very necessary that the ultramicro microscope imaging system can record double -color fluorescence. SUMMARY

[0003] The utility model solves the technical problem that the prior art has the defect, and the utility model provides a double -color ultramicro microscope imaging system that can effectively filter out the motion noise generated in the recording process.The ultramicro microscope imaging system of double -color excitation light can realize the synchronous recording and comparison of multiple fluorescent proteins, and the activity of two types of neurons in a behavior paradigm in the related brain area can be simultaneously recorded by using the system, so as to reflect the encoding characteristics of different types of neurons in the same behavior paradigm.

[0004] To solve the above technical problems, the utility model adopts the technical scheme of a double -color ultramicro microscope imaging system, which comprises a video capture card, a double -color ultramicro microscope imaging mirror body, a multi-core optical fiber bundle, an LED driving circuit, a digital signal acquisition card and a switching power supply, wherein the video capture card is electrically connected with the LED driving circuit, which is used to distinguish odd frames and even frames in units of image frames by communicating with computer acquisition software, and synchronously control the corresponding light source signal, the double -color ultramicro microscope imaging mirror body is used to emit two excitation lights and collect fluorescent images, the multi-core optical fiber bundle is used to couple two excitation lights into a light source and access the double -color ultramicro microscope imaging mirror body, the LED driving circuit is used to receive the control signal of the video capture card to realize the switching of excitation light and the synchronization of image frames, the digital signal acquisition card is electrically connected with the video capture card, which is used to receive the marking signal of external behavior, and realize synchronous analysis with image frame signal, and the switching power supply is electrically connected with the LED driving circuit.

[0005] Further, a case is further included, the video acquisition card, the double color ultramicroscopic imaging mirror body, the multi-core optical fiber bundle, the LED driving circuit, the digital signal acquisition card and the switching power supply are all assembled through the case.

[0006] Further, the LED driving circuit includes a driving chip, a PWM input module, a power module and an LED lamp bead module, the PWM input module is used to input the synchronization signal output by the external video acquisition card and is electrically connected with the driving chip, the power module and the LED lamp bead module are respectively electrically connected with the corresponding pins of the driving chip, the power module is internally provided with a 5V power supply, the driving chip is further electrically connected with a dimming resistor, and the dimming resistor is used to realize the strength control of the LED lamp bead module in cooperation with the PWM input module.

[0007] Further, a collection and analysis module is further included, the collection and analysis module is in communication with the video acquisition card, and is used to separately save different fluorescence signals and analyze the fluorescence change curve data of the corresponding single cells.

[0008] Further, in the collection and analysis module, the collection module realizes the differentiation of different color fluorescence signals by reading the data stored in the specific register of the video acquisition card, and the analysis module can correct the jitter noise generated in the free motion process.

[0009] Further, the collection module is a video synchronous collection software written based on a C# language, and the analysis module is an interface program written based on a MATLAB language.

[0010] Further, the multi-core optical fiber bundle can include a plurality of optical fibers coupled with different light sources.

[0011] Compared with the prior art, the ultramicroscopic imaging system can be widely applied to various fields, such as biomedical science, life science and the like, and achieves the following technical effects:

[0012] 1. The entire system can realize the synchronous recording of multiple fluorescence signals through time division multiplexing, and can realize the synchronous stimulation and recording at the single cell level at the same time;

[0013] 2. The design of the ultramicroscopic imaging system with different functions can be realized by adjusting the combination of the external excitation light source and the filter of the mirror body. BRIEF DESCRIPTION OF DRAWINGS

[0014] The disclosure of the present application will be described with reference to the accompanying drawings. It should be understood that the drawings are only for illustrative purposes, and are not intended to limit the scope of protection of the present application. In the drawings, the same reference numerals are used to refer to the same parts. Among them:

[0015] Fig. 1The machine case structure schematic diagram according to the embodiment of the utility model is shown schematically;

[0016] Fig. 2 The LED drive circuit diagram according to the embodiment of the utility model is shown schematically;

[0017] Marked number in the drawing: 1, machine case;2, drive chip;3, PWM input module;4, power module;5, LED lamp pearl module;6, light modulation resistance. DETAILED DESCRIPTION

[0018] It is easy to understand that according to the technical scheme of the utility model, the general skilled person in the art can propose a plurality of structure modes and implementation modes that can be mutually replaced 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 of the utility model or regarded as the limitation or restriction of the technical scheme of the utility model.

[0019] According to the embodiment of the utility model in combination Figs. 1-2 It is shown.

[0020] The utility model aims at providing a kind of double color ultramicro microscopic imaging system, by the combination of video capture card, double color ultramicro microscopic imaging mirror body, custom-made multi-core fiber bundle, LED drive circuit, digital signal acquisition card, switching power supply, machine case 1 and acquisition and analysis software, the synchronous recording of multiple fluorescence signals or single-cell level synchronous stimulation and recording are realized.

[0021] Specifically, in the embodiment, a kind of double color ultramicro microscopic imaging system, including video capture card, double color ultramicro microscopic imaging mirror body, custom-made multi-core fiber bundle, LED drive circuit, digital signal acquisition card, switching power supply, machine case 1 and acquisition and analysis software etc.;The internal program of video capture card carries out time division multiplexing to video data stream by image frame as unit, distinguishes odd frame and even frame;And corresponding light source signal is synchronously controlled, the synchronization and time division acquisition of light source and image frame are realized;Double color ultramicro microscopic imaging mirror body realizes the reflection of two excitation lights and the collection imaging function of fluorescence signal, the image of single cell emission is transmitted to computer by video capture card;Custom-made fiber bundle can couple two excitation lights into a light source and access double color ultramicro microscopic imaging mirror body;LED drive circuit can receive the control signal of video capture card and realize the switch of excitation light and the synchronization of video frame;Digital signal acquisition card is used to receive the marking signal of external behavior, realizes the synchronous analysis with image frame signal;Switching power supply can realize the power supply to LED drive circuit;Acquisition and analysis software can realize data communication with video capture card, different fluorescence signals are saved separately, and the fluorescence change curve data of corresponding single cell is analyzed.

[0022] Furthermore, the aforementioned dual-color ultra-miniature microscopic imaging system can be customized with multi-core fiber bundles that can be flexibly changed according to customer experimental needs, enabling the mirror to couple with different light sources.

[0023] With the above system setup, the acquisition software distinguishes different colored fluorescence signals by reading data stored in specific registers of the video acquisition card; the analysis software has a motion correction function, which can correct the jitter noise generated during free movement, thereby achieving accurate extraction of single-cell fluorescence signals.

[0024] For the specific implementation of the above driving circuit, such as Fig. 2 As shown, the LED driving circuit includes a driver chip 2, a PWM input module 3, a power supply module 4, and an LED bead module 5. The PWM input module 3 is used to input the synchronization signal output by the external video capture card and is electrically connected to the driver chip 2. The power supply module 4 and the LED bead module 5 are electrically connected to the corresponding pins of the driver chip 2. The power supply module 4 has a built-in 5V power supply. The driver chip 2 is also electrically connected to a dimming resistor 6. The dimming resistor 6 is used to cooperate with the PWM input module 3 to control the strength of the LED bead module 5.

[0025] The dual-color ultra-miniature microscopy imaging system in this scheme needs to achieve simultaneous recording of multicolor fluorescence without increasing the weight and volume of the microscope. Furthermore, due to the overlapping fluorescence emission spectra of the two excitation lights, the system must employ time-division multiplexing technology to achieve simultaneous recording of the two fluorescencees. One dual-color ultra-miniature microscopy imaging system places the two excitation lights externally within the microscope body, and guides different colors of excitation light into the system via a customized multi-core fiber bundle. Simultaneously, the video data stream is time-division multiplexed frame by frame using the internal program of the video acquisition card to distinguish between odd and even frames. The corresponding light source signals are synchronously controlled to achieve synchronization and time-division acquisition of the light source and image frames. Based on these two technical solutions, simultaneous recording of two fluorescencees is achieved without increasing the weight and volume of the ultra-miniature microscopy imaging system. Additionally, this system can replace one of the excitation lights with a high-power red laser to achieve synchronous stimulation and recording at the single-cell level.

[0026] The technical scope of this utility model is not limited to the content described above. Those skilled in the art can make various modifications and variations to the above embodiments without departing from the technical concept of this utility model, and all such modifications and variations should fall within the protection scope of this utility model.

Claims

1. A two-color ultramicroscopic imaging system, characterized by, It includes: Video capture card, double color super micro imaging mirror, multi-core optical fiber bundle, LED drive circuit, digital signal acquisition card and switching power supply, wherein the video capture card is electrically connected with the LED drive circuit, which is used to distinguish odd and even frames by communicating with computer acquisition software in units of image frames, and synchronously control the corresponding light source signal, the double color super micro imaging mirror is used to collect and image the emission of two excitation lights and the collection of fluorescence model, the multi-core optical fiber bundle is used to couple two excitation lights into a light source and access to the double color super micro imaging mirror, the LED drive circuit is used to receive the control signal of the video capture card to realize the synchronization of the excitation light and the image frame, the digital signal acquisition card is electrically connected with the video capture card, which is used to receive the external behavioral marking signal for synchronous analysis with the image frame signal; the switching power supply is electrically connected with the LED drive circuit.

2. The dual-color ultramicroscopy imaging system of claim 1, wherein: It also includes a case, and the video capture card, double color super micro imaging mirror, multi-core optical fiber bundle, LED drive circuit, digital signal acquisition card and switching power supply are assembled in the case.

3. The dual-color ultramicroscopy imaging system of claim 1, wherein: The LED drive circuit includes a drive chip, a PWM input module, a power module and an LED lamp bead module, the PWM input module is used to input the synchronization signal output by the external video capture card and is electrically connected with the drive chip, the power module and the LED lamp bead module are respectively connected with the corresponding pins of the drive chip, the power module is built-in 5V power supply, the drive chip is also electrically connected with a dimming resistor, and the dimming resistor is used to realize the strong and weak control of the LED lamp bead module in cooperation with the PWM input module.

4. The dual-color ultramicroscopy imaging system of claim 1, wherein: The multi-core optical fiber bundle can include a plurality of different light sources.

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