CN110584616A - Dual-mode imaging microscope system - Google Patents
Dual-mode imaging microscope system Download PDFInfo
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- CN110584616A CN110584616A CN201910994586.0A CN201910994586A CN110584616A CN 110584616 A CN110584616 A CN 110584616A CN 201910994586 A CN201910994586 A CN 201910994586A CN 110584616 A CN110584616 A CN 110584616A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0033—Features or image-related aspects of imaging apparatus classified in A61B5/00, e.g. for MRI, optical tomography or impedance tomography apparatus; arrangements of imaging apparatus in a room
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0033—Features or image-related aspects of imaging apparatus classified in A61B5/00, e.g. for MRI, optical tomography or impedance tomography apparatus; arrangements of imaging apparatus in a room
- A61B5/004—Features or image-related aspects of imaging apparatus classified in A61B5/00, e.g. for MRI, optical tomography or impedance tomography apparatus; arrangements of imaging apparatus in a room adapted for image acquisition of a particular organ or body part
- A61B5/0042—Features or image-related aspects of imaging apparatus classified in A61B5/00, e.g. for MRI, optical tomography or impedance tomography apparatus; arrangements of imaging apparatus in a room adapted for image acquisition of a particular organ or body part for the brain
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
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- A—HUMAN NECESSITIES
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- A61B5/0093—Detecting, measuring or recording by applying one single type of energy and measuring its conversion into another type of energy
- A61B5/0095—Detecting, measuring or recording by applying one single type of energy and measuring its conversion into another type of energy by applying light and detecting acoustic waves, i.e. photoacoustic measurements
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- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6801—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be attached to or worn on the body surface
- A61B5/6802—Sensor mounted on worn items
- A61B5/6803—Head-worn items, e.g. helmets, masks, headphones or goggles
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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Abstract
The embodiment of the invention provides a dual-mode imaging microscope system, which comprises: the device comprises a light source module, a lens group module, an upper computer, an optical fiber transmission module, a conductive slip ring and a mobile microscope. The upper computer controls the light source module to emit laser, and the laser is transmitted to the mobile microscope through the lens group module and the optical fiber transmission module penetrating through the conductive slip ring through hole; the upper computer controls the laser to scan the target object according to a preset path so as to excite a fluorescence signal and a photoacoustic signal; the fluorescence signal and the photoacoustic signal are converted into electric signals and then transmitted to an upper computer; and the upper computer performs real-time imaging according to the electric signal. The embodiment of the invention adopts the photoacoustic imaging technology and the fluorescence imaging technology, utilizes the conductive slip ring to connect the mobile microscope and other equipment, realizes multifunctional imaging in the moving state of the target object, avoids the wire winding caused by the movement of the target object when the multifunctional imaging of the target object is carried out, has compact structure, realizes the miniaturization of the system and realizes the movable wearing of the imaging microscope.
Description
Technical Field
The embodiment of the invention relates to the technical field of microscopic imaging, in particular to a dual-mode imaging microscope system.
Background
At present, brain imaging modes studied at home and abroad mainly comprise nuclear magnetic resonance imaging, CT imaging, electroencephalogram imaging, optical imaging, photoacoustic imaging and other imaging modes. Among them, magnetic resonance imaging and CT imaging have been widely used in clinical practice, but the time resolution is insufficient and there is a potential radiation hazard to human body; electroencephalogram imaging is the most direct imaging mode for observing neuroelectrical activity in brain science research, but the spatial resolution is not sufficient. Optical imaging and photoacoustic imaging are mainly used in animal basic research at present, and for animal cerebrovascular network research, a single/two-photon microscope in an optical imaging method has achieved wearable imaging of a mouse, however, the imaging range is small, the depth is shallow, and the brain microenvironment may be damaged by injection of fluorescent dye. The photoacoustic imaging combines the advantages of optical imaging and ultrasonic imaging, not only has higher resolution, but also has deeper imaging depth. However, the conventional optical microscopic imaging system has a complicated optical path, and the conventional mechanical scanning has high precision requirement on a motor, a large size, a low speed and a high price, and cannot realize miniaturization and rapid imaging of the system, so that the conventional optical microscopic imaging system always stays in an animal brain imaging state or a body fixing state, and cannot perform real-time imaging on a cerebral vascular network of a rat in a free moving state. In addition, the imaging mode is single, so that the traditional brain imaging cannot link the hemodynamic change and the neuroelectric activity dynamics in the brain activity process, and the structural and functional information of the brain is not completely known.
In order to solve the problems, some groups at home and abroad propose a method based on a high-performance voice coil motor, which combines optical scanning with mechanical scanning and uses a micro scanning electron microscope, so that the imaging speed is effectively improved, the system volume is reduced to a certain extent, the system use portability is improved, and the movable wearing of the system and the multifunctional imaging of animal brain function activities cannot be realized.
Disclosure of Invention
In view of this, the embodiment of the present invention provides a dual-mode imaging microscope system to solve the problem of how to implement multifunctional imaging of an animal brain in a free movement state.
The embodiment of the invention provides a dual-mode imaging microscope system which is used for an animal brain multifunctional imaging process in a free moving state. The dual-mode imaging microscope system comprises: the device comprises a light source module, a lens group module, an upper computer, an optical fiber transmission module, a conductive slip ring and a mobile microscope;
the upper computer is used for generating a laser emission control signal and a laser scanning control signal, and the laser scanning control signal is transmitted to the mobile microscope through a first lead communicated with the conductive slip ring;
the light source module is used for emitting laser according to the laser emission control signal, and the laser enters the optical fiber transmission module through the lens group module;
the optical fiber transmission module penetrates through the through hole of the conductive slip ring and then is connected with the mobile microscope so as to transmit the laser to the mobile microscope, and the mobile microscope controls the laser to scan the surface of a target object according to a preset path according to the laser scanning control signal so as to excite a fluorescence signal and a photoacoustic signal;
the mobile microscope returns the fluorescence signal to the lens group module through the optical fiber transmission module, and the fluorescence signal is photoelectrically converted into a first electric signal and then transmitted to the upper computer;
and the mobile microscope converts the photoacoustic signal into a second electric signal, and the second electric signal is transmitted to the upper computer through a second lead communicated with the conductive slip ring.
Preferably, the light source module includes: a first laser generator and a second laser generator;
the first laser generator generates first laser light and the second laser generator generates second laser light.
Further, the lens group module includes:
and the first dichroic mirror is used for transmitting the first laser and reflecting the second laser so as to realize coaxial transmission of the first laser and the second laser.
Further, the host computer includes:
the photoelectric conversion module is used for converting the fluorescence signal into the first electric signal;
the data acquisition card is used for acquiring the first electric signal and the second electric signal, and the upper computer of the first laser generator displays a two-dimensional image in real time;
and the laser scanning signal generator is used for outputting the laser scanning control signal.
Specifically, the lens group module includes:
the second dichroic mirror is used for transmitting the laser and reflecting the fluorescence signal, and separating the fluorescence signal returned to the lens group module from the original laser transmission light path to the photoelectric conversion module;
the photoelectric conversion module comprises a photomultiplier tube and is used for converting the fluorescence signal into the first electric signal.
Specifically, the optical fiber transmission module includes: the optical fiber coupler, the rotary optical fiber and the multimode optical fiber rotary connector;
the optical fiber coupler is connected with the rotating optical fiber and is used for coupling the laser into the rotating optical fiber;
the multimode optical fiber rotary connector internally fixes the rotary optical fiber;
the multimode optical fiber rotary connector is fixed at the through hole of the conductive slip ring.
Preferably, the lens group module includes:
the spatial filter is used for adjusting the laser into a collimated Gaussian beam;
and the first focusing lens is used for focusing the collimated Gaussian beam and transmitting the collimated Gaussian beam to the mobile microscope through the optical fiber transmission module.
Specifically, the mobile microscope includes:
the reflecting mirror surface of the scanning galvanometer receives the laser output by the optical fiber transmission module, and the scanning galvanometer is used for controlling the laser to scan the surface of the target object according to a preset path according to the laser scanning control signal;
an ultrasound probe for converting the photoacoustic signal into a second electrical signal;
and the light transmission anti-sound device is used for transmitting the fluorescence signal and reflecting the photoacoustic signal, returning the fluorescence signal to the lens group module in the reverse original laser transmission direction by the first laser generator, and reflecting the photoacoustic signal to the ultrasonic detector.
Further, the mobile microscope further comprises: a variable focus collimating lens and a second focusing lens;
the variable-focus collimating lens is used for collimating the laser transmitted to the mobile microscope;
the second focusing lens can focus the collimated laser on the reflecting mirror surface of the scanning galvanometer.
Specifically, the host computer includes:
the first data acquisition card is used for acquiring the first electric signal and the second electric signal, and the upper computer of the first laser generator displays a two-dimensional image in real time;
and the laser scanning signal generator is used for outputting the laser scanning control signal.
Further, the dual-mode imaging microscope system further comprises a micro-conversion bracket for fixing the mobile microscope to the target object.
The working process of the embodiment of the invention is as follows:
the upper computer generates a laser emission control signal to control the light source module to emit laser, and the laser enters the mobile microscope through the lens group module and the optical fiber transmission module penetrating through the conductive slip ring through hole; meanwhile, the upper computer also generates a laser scanning control signal, the laser scanning control signal enters the mobile microscope through a first lead communicated with the conductive slip ring, and the mobile microscope controls laser to scan the surface of the target object according to a preset path according to the laser scanning control signal so as to excite a fluorescence signal and a photoacoustic signal; the mobile microscope can return the fluorescent signal to the lens group module through the optical fiber transmission module, and the fluorescent signal is photoelectrically converted into a first electric signal and then transmitted to an upper computer; meanwhile, the mobile microscope also converts the photoacoustic signal into a second electrical signal, and the second electrical signal is transmitted to an upper computer through a second lead communicated with the conductive slip ring; the upper computer analyzes the first electric signal and the second electric signal and displays the two-dimensional image in real time, and multifunctional imaging of the target object in the free moving state is achieved.
The embodiment of the invention adopts the photoacoustic imaging technology and the fluorescence imaging technology, utilizes the conductive slip ring to connect the mobile microscope and other equipment, realizes multifunctional imaging in the moving state of a target object, realizes the joint research of a brain activity mechanism from two aspects of cranial nerve activity and hemodynamics, and is favorable for more comprehensively and deeply researching the brain activity mechanism; the problem that the electric wire is wound due to the fact that the target object moves when multifunctional imaging of the target object is carried out is avoided; the imaging microscope has small volume and light weight, can realize brain imaging in a wearable free moving state, is closer to the normal physiological state of organisms, and breaks through the limitation that anesthesia and fixation are needed in the traditional brain microscopic imaging, thereby widening the application range of the brain microscopic imaging technology and expanding the research range of the brain microscopic imaging; the imaging quality is high, the speed is high, the imaging performance is good, the cerebrovascular morphological structure can be observed more visually, the cerebral cortex vascular network can be known, and the deep research and development of animal brain science can be facilitated.
Drawings
Fig. 1 is a schematic structural diagram of a dual-mode imaging microscope system according to a first embodiment of the present invention;
fig. 2 is a schematic path diagram of a preset scanning path in the first embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a dual-mode imaging microscope system according to a second embodiment of the present invention;
FIG. 4 is a schematic structural diagram of a dual-mode imaging microscope system in an alternative embodiment of the present invention;
FIG. 5 is a schematic structural diagram of a mobile microscope according to a third embodiment of the present invention;
fig. 6 is an image of photoacoustic imaging results of rat blood vessels obtained by the third embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention. It should be further noted that, for the convenience of description, only some of the structures related to the present invention are shown in the drawings, not all of the structures.
Example one
Fig. 1 is a schematic structural diagram of a dual-mode imaging microscope system according to an embodiment of the present invention. As shown in fig. 1, an embodiment of the present invention provides a dual-mode imaging microscope system, which includes an upper computer 1, a light source module 2, a lens group module 3, an optical fiber transmission module 4, a conductive slip ring 5, and a mobile microscope 6. The dual-mode imaging microscope system of the present embodiment is used to image the target object 7.
The upper computer 1 may generate a laser emission control signal and a laser scanning control signal.
Specifically, the laser scanning control signal may be transmitted to the mobile microscope 6 through a first wire communicated by the conductive slip ring 5; the laser emission control signal is transmitted to the light source module to control the light source module 2 to emit laser.
The light source module 2 emits laser according to a laser emission control signal generated by the upper computer 1, and the laser enters the optical fiber transmission module 4 through the lens group module 3.
The optical fiber transmission module 4 is connected to the mobile microscope 6 after penetrating through the through hole of the conductive slip ring 5 to transmit laser to the mobile microscope 6, and the mobile microscope 6 can control the laser to scan the surface of the target object 7 according to a preset path according to a laser scanning control signal generated by the upper computer 1 to excite a fluorescence signal and a photoacoustic signal. The preset path may be, as shown by an arrow in fig. 2, raster-scan the target object 7 along the imaging area.
Specifically, the optical fiber transmission module 4 may include an optical fiber coupler and a rotating optical fiber, and the optical fiber coupler is connected to the rotating optical fiber, and may couple the laser into the rotating optical fiber to be transmitted to the mobile microscope 6. The rotary optical fiber can be divided into a front-end rotary optical fiber and a rear-end rotary optical fiber which are fixed at the through hole of the conductive slip ring through the multimode optical fiber rotary connector. For example, the front end rotating optical fiber may be fixed to a stator of the multimode optical fiber rotary connector, and the rear end rotating optical fiber may be fixed to a rotor of the multimode optical fiber rotary connector, so that when the target object 7 moves to drive the rear end rotating optical fiber to rotate, the rotor and the stator are fixed, and still remain coaxial with the front end rotating optical fiber, thereby avoiding interruption of optical signal transmission caused by optical fiber rotation when the target object moves.
Further, the mobile microscope 6 can return the fluorescence signal to the lens group module 3 through the optical fiber transmission module 4, and then the fluorescence signal is photoelectrically converted into a first electric signal and transmitted to the upper computer 1.
Specifically, due to the reversibility of the optical path, a fluorescence signal excited after the laser irradiates the target object can return to the lens group module along the reverse original optical path transmission direction; the upper computer 1 can be connected with a photomultiplier to convert a fluorescent signal into a first electric signal.
Further, the movable microscope 6 can convert the photoacoustic signal into a second electrical signal, which is transmitted to the upper computer 1 through a second wire communicated by the conductive slip ring 5.
In particular, the conversion of the photoacoustic signal into the second electrical signal may be achieved by an ultrasound probe built into the mobile microscope 6.
When the dual-mode imaging microscope system provided by the embodiment of the invention works, the upper computer 1 generates a laser emission control signal to control the light source module 2 to emit laser, and then the laser enters the movable microscope 6 through the lens group module 3 and the optical fiber transmission module 4 penetrating through the through hole of the conductive slip ring 5; meanwhile, the upper computer 1 also generates a laser scanning control signal, the laser scanning control signal enters the mobile microscope 6 through a first lead communicated with the conductive slip ring 5, and the mobile microscope 6 controls laser to scan the surface of the target object 7 according to a preset path according to the laser scanning control signal so as to excite a fluorescence signal and a photoacoustic signal; the mobile microscope 6 can return the fluorescence signal to the lens group module 3 through the optical fiber transmission module 4, and the fluorescence signal is photoelectrically converted into a first electric signal and then transmitted to the upper computer 1; meanwhile, the mobile microscope 6 also converts the photoacoustic signal into a second electrical signal, and the second electrical signal is transmitted to the upper computer 1 through a second wire communicated with the conductive slip ring 5; the upper computer 1 analyzes the first electric signal and the second electric signal and displays the two-dimensional image in real time, thereby realizing multifunctional imaging of the target object 7 in a free moving state. The target object of this embodiment may be the surface of an animal brain, such as the brain of an experimental rat.
The embodiment of the invention adopts the photoacoustic imaging technology and the fluorescence imaging technology, utilizes the conductive slip ring to connect the mobile microscope and other equipment, realizes the multifunctional imaging in the moving state of the target object, avoids the winding of electric wires caused by the movement of the target object when the multifunctional imaging of the target object is carried out, has simple structural design and compact connection, realizes the miniaturization of a system, realizes the mobile wearable imaging of the imaging microscope, and is beneficial to the deep research and development of animal brain science.
Example two
Fig. 3 is a schematic structural diagram of a dual-mode imaging microscope system in a second embodiment of the present invention, which can implement photoacoustic imaging and fluorescence imaging of a target object in a free moving state, and as shown in fig. 3, with reference to fig. 1, on the basis of the first embodiment, a light source module 2 in the second embodiment of the present invention includes: a first laser generator 21 and a second laser generator 22.
Specifically, the first laser generator 21 generates a first laser, and the second laser generator 22 generates a second laser, wherein the first laser may be a pulse laser in a visible light or near infrared band, and the second laser may be a pulse laser in an ultraviolet band. Preferably, the first laser light is pulse laser light with a wavelength of 532nm, and the second laser light is pulse laser light with a wavelength of 577 nm.
Further, lens group module 3 includes first dichroic mirror 31, and the parameters of first dichroic mirror 31 are not specifically limited, and only needs to transmit first laser and reflect second laser to realize coaxial transmission of first laser and second laser.
Preferably, lens group module 3 further includes a reflecting mirror 33, and reflecting mirror 33 can reflect the second laser light emitted by second laser generator 22 to first dichroic mirror 31, so as to implement coaxial transmission of the second laser light and the first laser light through control of first dichroic mirror 31.
Specifically, the upper computer 1 includes: a photoelectric conversion module 11, a data acquisition card 12 and a laser scanning signal generator 13. The photoelectric conversion module 11 can receive the fluorescence signal returned to the lens group module 3 and convert the fluorescence signal into a first electric signal; the data acquisition card 12 can acquire a first electric signal and a second electric signal obtained by converting the photoacoustic signal through the mobile microscope 6, and the upper computer 1 of the first laser generator displays a two-dimensional image in real time; the laser scanning signal generator 13 can output a laser scanning control signal according to an instruction of the upper computer 1, and transmits the laser scanning control signal to the mobile microscope 6 through the conductive slip ring 5.
Preferably, the photoelectric conversion module 11 is a photomultiplier tube, and can detect a weak fluorescent signal and convert it into a first electrical signal.
Preferably, the upper computer 1 is internally provided with Labview software, the Labview software controls the data acquisition card 12 to continuously acquire and process a first electrical signal and a second electrical signal of original three-dimensional image data containing the target object 7, and the acquired first electrical signal is restored and the second electrical signal is subjected to maximum projection, so that the upper computer 1 can display a two-dimensional image in real time. Image processing steps such as image noise removal, contrast enhancement, vascularization parameter extraction and the like can be performed through Matlab software.
Preferably, the laser scanning signal generator 13 is a function generator, and the function generator can be controlled by Labview software built in the upper computer 1 to generate a laser scanning control signal. The traveling microscope 6 controls the laser to scan the surface of the target object 7 in a preset path as shown in fig. 2 according to the laser scanning control signal to excite the fluorescence signal and the photoacoustic signal.
Further, the lens group module further includes a second dichroic mirror 32. Preferably, second dichroic mirror 32 is coaxial with first dichroic mirror 31, and parameters of first dichroic mirror 31 are not specifically limited, but only first laser and second laser are transmitted and reflected to separate a fluorescence signal returned to lens group module 3 from an original laser transmission light path to photoelectric conversion module 11.
Further, the fiber transmission module 4 includes a fiber coupler 41 and a rotating fiber 42.
Specifically, the fiber coupler 41 is connected to the rotating fiber 42, and couples the laser light into the rotating fiber 42 and transmits the laser light to the movable microscope 6.
Preferably, the rotating optical fiber 42 can be divided into a front rotating optical fiber and a rear rotating optical fiber, and is fixed at the through hole of the conductive slip ring 5 through the multimode optical fiber rotating connector 43. For example, the front end rotating optical fiber may be fixed to a stator of the multimode optical fiber rotating connector 43, and the rear end rotating optical fiber may be fixed to a rotor of the multimode optical fiber rotating connector 43, so that when the target object 7 moves to drive the rear end rotating optical fiber to rotate, the rear end rotating optical fiber still remains coaxial with the front end rotating optical fiber due to the fixation of the axial positions of the rotor and the stator, thereby avoiding the interruption of optical signal transmission caused by the rotation of the optical fiber when the target object moves. Further, the rear-end rotating optical fiber, a part of the first wire for connecting the conductive slip ring 5 and the mobile microscope 6, and a part of the second wire for connecting the conductive slip ring 5 and the mobile microscope 6 are bound together to form a hybrid connecting wire 8.
When the dual-mode imaging microscope system provided by the embodiment of the invention works, the upper computer 1 generates a laser emission control signal, so that the first laser generator 21 generates first laser and the second laser generator 22 generates second laser, then the first laser and the second laser form mixed light beams through the first dichroic mirror 31, and the mixed light beams pass through the second dichroic mirror 32, enter the rotating optical fiber 42 through the optical fiber coupler 41, and then enter the mobile microscope 6; meanwhile, Labview software is arranged in the upper computer 1, the Labview software can control a laser scanning signal generator to generate a laser scanning control signal, and the mobile microscope 6 controls laser to scan the surface of the target object 7 according to a preset path according to the laser scanning control signal so as to excite a fluorescence signal and a photoacoustic signal; the mobile microscope 6 can return the fluorescence signal to the lens group module 3 through the optical fiber transmission module 4, and then reflect the fluorescence signal to the photoelectric conversion module 11 through the second dichroic mirror 32. The fluorescence signal is converted into a first electrical signal by the photoelectric conversion module 11 and then enters the data acquisition card 12. Meanwhile, the mobile microscope 6 may convert the photoacoustic signal excited by the target object 7 into a second electrical signal, and transmit the second electrical signal to the data acquisition card 12. Labview software is arranged in the upper computer 1, the data acquisition card 12 is controlled by the Labview software to continuously acquire and process a first electric signal and a second electric signal of original three-dimensional image data containing the target object 7, and the acquired first electric signal and the acquired second electric signal are subjected to maximum value projection, so that the upper computer 1 can display a two-dimensional image in real time. Multi-functional imaging of the target object 7 in the freely moving state is realized.
The embodiment of the invention adopts a photoacoustic imaging technology and a fluorescence imaging technology, utilizes the conductive slip ring to connect the mobile microscope and other equipment, and utilizes the rotating optical fiber to enable laser to enter the mobile microscope, thereby realizing multifunctional imaging in a moving state of a target object. Through adopting the portable microscope of conductive slip ring connection and other equipment and adopting the rotatory optic fibre transmission laser that is fixed in optic fibre rotary connector, avoided when carrying out the multi-functional formation of image of target object the target object removes and leads to the electric wire winding, simultaneously, structural design is simple, connects compactly, has realized the miniaturization of system, has realized that imaging microscope's movable is worn the ization, is favorable to the deep research development of animal brain science.
In an alternative embodiment, as shown in fig. 4, lens module 3 further includes a spatial filter 34 disposed behind first dichroic mirror 31 for adjusting the mixed beam of the first laser light and the second laser light into a collimated gaussian beam.
The lens assembly module 3 further comprises a first focusing lens 35 disposed behind the spatial filter 34 for focusing the collimated gaussian beam onto the fiber coupler and into the rotating fiber 42 through the fiber coupler 41.
Preferably, the spatial filter 34 may be constituted by the objective lens 341 in combination with the aperture stop 342.
Preferably, when the fluorescent signal passes through the light-transmitting anti-sound device 65 and returns to the second dichroic mirror 32 in the reverse original optical path transmission direction and is reflected to the photoelectric conversion module 11 by the second dichroic mirror 32, the fluorescent signal also passes through a band-pass filter 36, and the band-pass filter 36 can filter out other stray light contained in the reflected fluorescent signal, so that only the fluorescent signal to be detected is left, and the influence of the stray light on the imaging result is avoided.
Preferably, when the first electrical signal is transmitted to the data acquisition card 12, the first electrical signal may be further amplified by the signal amplifier 14, and noise is filtered out by the built-in capacitor bank, so as to obtain the first electrical signal with good signal-to-noise ratio, and obtain better imaging result.
EXAMPLE III
Fig. 5 is a diagram of an embodiment of a mobile microscope structure in a third embodiment of the present invention, which is a further refinement of the mobile microscope structure in any embodiment of the present invention, and referring to fig. 3, specifically, the mobile microscope 6 includes: a variable focus collimating lens 61, a second focusing lens 63, an ultrasound probe 64, a light transmissive anti-sound device 65 and a scanning galvanometer 66.
Preferably, the scanning galvanometer 66 is a micro-electromechanical scanning galvanometer to reduce the mass of the mobile microscope 6.
Preferably, the mobile microscope 6 further includes a right-angle prism 62 in the embodiment of the present invention to change the transmission direction of the light path, so that the mobile microscope 6 is more compact. The variable focus collimating lens 61 receives the laser light from the optical fiber transmission module 4, and adjusts the laser light into collimated laser light, and then the collimated laser light is reflected by the right angle prism 62 and focused on the reflecting mirror surface of the scanning galvanometer 66 by the second focusing lens 63. The reflecting mirror surface of the scanning galvanometer 66 receives the laser and controls the scanning galvanometer 66 to rotate according to the laser scanning control signal generated by the upper computer 1 so as to control the laser to scan the target object 7 after being transmitted by the light-transmitting anti-sound device 65 according to a preset path.
Further, the target object 7 excites and reflects the fluorescence signal and the photoacoustic signal, after the fluorescence signal and the photoacoustic signal pass through the light-transmitting anti-sound device 65, the fluorescence signal passes through the light-transmitting anti-sound device 65 and returns to the lens group module 3 in the direction opposite to the original light path, and the photoacoustic signal is reflected to the ultrasonic detector 64 through the light-transmitting anti-sound device.
Further, the ultrasound probe 64 may convert the photoacoustic signal into a second electrical signal.
Further, in the dual-mode imaging microscope system provided in the embodiment of the present invention, a micro-switching bracket is further included for fixing the mobile microscope 6 to the target object.
Specifically, the micro conversion cradle may be made of metal, and only a high fit between the target object, such as the head of the rat, and the mobile microscope 6 is achieved, and the specific shape thereof is not more specifically limited.
When the mobile microscope provided by the embodiment of the invention is applied to the dual-mode imaging microscope system provided by any embodiment of the invention, in operation, the upper computer 1 generates a laser emission control signal to enable the first laser generator 21 to generate first laser and the second laser generator 22 to generate second laser, and then the first laser and the second laser form a mixed light beam through the first dichroic mirror 31, pass through the second dichroic mirror 32, enter the rotary optical fiber 42 through the optical fiber coupler 41, and then enter the mobile microscope 6; meanwhile, Labview software is arranged in the upper computer 1, the Labview software can control a laser scanning signal generator to generate a laser scanning control signal, and the mobile microscope 6 controls the scanning galvanometer 66 to rotate according to the laser scanning control signal so as to control laser to scan the surface of the target object 7 according to a preset path to excite a fluorescence signal and a photoacoustic signal; the mobile microscope 6 can return the fluorescence signal to the lens group module 3 through the optical fiber transmission module 4, and then reflect the fluorescence signal to the photoelectric conversion module 11 through the second dichroic mirror 32. The fluorescence signal is converted into a first electrical signal by the photoelectric conversion module 11 and then enters the data acquisition card 12. Meanwhile, the ultrasonic detector 64 may convert the photoacoustic signal excited by the target object 7 into a second electrical signal and transmit the second electrical signal to the data acquisition card 12. Labview software is arranged in the upper computer 1, the data acquisition card 12 is controlled by the Labview software to continuously acquire and process a first electric signal and a second electric signal of original three-dimensional image data containing the target object 7, and the acquired first electric signal and the acquired second electric signal are subjected to maximum value projection, so that the upper computer 1 can display a two-dimensional image in real time. Multi-functional imaging of the target object 7 in the freely moving state is realized. When the dual-mode imaging microscope system in the embodiment of the invention is used for imaging the rat cerebrovascular network in a free moving state, the imaging range is 1.2mm multiplied by 1.2mm, the acquisition frequency is 5Hz, and the rapid raster scanning is carried out on the imaging area of the target object by only using the first laser with the output wavelength of 532nm, the pulse width of 2ns and the repetition frequency of 50KHz, so that the cerebrovascular network structural image shown in figure 6 is obtained.
The embodiment of the invention adopts a photoacoustic imaging technology and a fluorescence imaging technology, utilizes the conductive slip ring to connect the mobile microscope and other equipment, and utilizes the rotating optical fiber to enable laser to enter the mobile microscope, thereby realizing multifunctional imaging in a moving state of a target object. By adopting the conductive slip ring to connect the mobile microscope and other equipment and adopting the rotary optical fiber fixed on the optical fiber rotary connector to transmit laser, the electric wire winding caused by the movement of the target object when the multifunctional imaging of the target object is carried out is avoided. In addition, the variable-focus collimating lens 61 can be suitable for brain imaging of different planes, and meanwhile, elements in the movable microscope are all light elements, so that the movable microscope is simple in structural design, compact in connection and easy to wear, miniaturization of a system is realized, movable wearing of the imaging microscope is realized, and deep research and development of animal brain science are facilitated.
The foregoing is considered as illustrative only of the preferred embodiments of the invention and is for the purpose of promoting an understanding of the principles of the invention and is to be understood that the scope of the invention is not limited by this specific disclosure. All such possible equivalents and modifications are deemed to fall within the scope of the invention as defined in the appended claims.
Claims (10)
1. A dual-mode imaging microscope system is characterized by comprising a light source module, a lens group module, an upper computer, an optical fiber transmission module, a conductive slip ring and a movable microscope;
the upper computer is used for generating a laser emission control signal and a laser scanning control signal, and the laser scanning control signal is transmitted to the mobile microscope through a first lead communicated with the conductive slip ring;
the light source module emits laser according to the laser emission control signal, and the laser enters the optical fiber transmission module through the lens group module;
the optical fiber transmission module penetrates through the through hole of the conductive slip ring and then is connected with the mobile microscope so as to transmit the laser to the mobile microscope, and the mobile microscope controls the laser to scan the surface of a target object according to a preset path according to the laser scanning control signal so as to excite a fluorescence signal and a photoacoustic signal;
the mobile microscope returns the fluorescence signal to the lens group module through the optical fiber transmission module, and the fluorescence signal is photoelectrically converted into a first electric signal and then transmitted to the upper computer;
and the mobile microscope converts the photoacoustic signal into a second electric signal, and the second electric signal is transmitted to the upper computer through a second lead communicated with the conductive slip ring.
2. The dual-mode imaging microscope system of claim 1, wherein the light source module comprises: a first laser generator and a second laser generator;
the first laser generator generates first laser light and the second laser generator generates second laser light.
3. The dual-mode imaging microscope system of claim 2, wherein the lens group module comprises:
and the first dichroic mirror is used for transmitting the first laser and reflecting the second laser so as to realize coaxial transmission of the first laser and the second laser.
4. The dual-mode imaging microscope system of claim 1, wherein the upper computer comprises:
the photoelectric conversion module is used for converting the fluorescence signal into the first electric signal;
the data acquisition card is used for acquiring the first electric signal and the second electric signal, and the upper computer of the first laser generator displays a two-dimensional image in real time;
and the laser scanning signal generator is used for outputting the laser scanning control signal.
5. The dual-mode imaging microscope system of claim 4, wherein the lens group module comprises:
the second dichroic mirror is used for transmitting the laser and reflecting the fluorescence signal, and separating the fluorescence signal returned to the lens group module from the original laser transmission light path to the photoelectric conversion module;
the photoelectric conversion module comprises a photomultiplier tube and is used for converting the fluorescence signal into the first electric signal.
6. The dual-mode imaging microscope system of claim 1, wherein the fiber optic transmission module comprises: the optical fiber coupler, the rotary optical fiber and the multimode optical fiber rotary connector are arranged on the optical fiber coupler;
the optical fiber coupler is connected with the rotating optical fiber and is used for coupling the laser into the rotating optical fiber;
the multimode optical fiber rotary connector internally fixes the rotary optical fiber;
the multimode optical fiber rotary connector is fixed at the through hole of the conductive slip ring.
7. The dual-mode imaging microscope system of claim 1, wherein the lens group module comprises:
the spatial filter is used for adjusting the laser into a collimated Gaussian beam;
and the first focusing lens is used for focusing the collimated Gaussian beam and transmitting the collimated Gaussian beam to the mobile microscope through the optical fiber transmission module.
8. The dual-mode imaging microscope system of claim 1, wherein the mobile microscope comprises:
the reflecting mirror surface of the scanning galvanometer receives the laser output by the optical fiber transmission module, and the scanning galvanometer is used for controlling the laser to scan the surface of the target object according to a preset path according to the laser scanning control signal;
an ultrasound probe for converting the photoacoustic signal into a second electrical signal;
and the light transmission anti-sound device is used for transmitting the fluorescence signal and reflecting the photoacoustic signal, returning the fluorescence signal to the lens group module in the reverse original laser transmission direction by the first laser generator, and reflecting the photoacoustic signal to the ultrasonic detector.
9. The dual-mode imaging microscope system of claim 8, wherein the mobile microscope further comprises: a variable focus collimating lens and a second focusing lens;
the variable-focus collimating lens is used for collimating the laser transmitted to the mobile microscope;
the second focusing lens can focus the collimated laser on the reflecting mirror surface of the scanning galvanometer.
10. The dual-mode imaging microscope system of claim 1, further comprising a micro-conversion mount for securing the mobile microscope to the target object.
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Application publication date: 20191220 |