A back-carried visualizing in-vivo calcium imaging device

Abstract

The application discloses a back-carrying visual in-vivo calcium imaging device and belongs to the technical field of in-vivo calcium imaging. The device comprises a device main body, a spine holder and fixing screws. The device main body comprises a track pulley system, four universal wheels and a spine holder base. The track pulley system is composed of two mutually perpendicular metal rods, namely a horizontal track and a vertical track. The bottom end of the vertical track is provided with a microscope fixing platform. The spine holder is connected with the spine holder base of the device main body through four fixing screws and is located below and inside the device main body. The mouse back-carrying visual in-vivo calcium imaging device provided by the application can fix the mouse spine and is not easy to move. The device can be combined with the device main body carrying an ultramicroscope. The device is ingenious in design, simple and reasonable in structure, stable in clamping, good in flexibility, convenient to use and strong in compatibility, and can make the calcium signal recording means more abundant.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of in vivo calcium imaging technology, in particular to a back-mounted visual in vivo calcium imaging device. BACKGROUND

[0002] Calcium imaging refers to a method for monitoring calcium ion concentration in tissues using calcium ion indicators. In the field of neural system research, calcium imaging technology is widely used in in vivo or in vitro experiments to monitor the changes of calcium ions in hundreds or thousands of neurons at the same time, so as to detect the activity of neurons. This technology has been pursued by neuroscientists all over the world since its inception, and it is still the most direct means for observing neuron activity.

[0003] In recent years, with the development of optogenetics and in vivo calcium imaging technology, people have a very perfect application means for brain in vivo calcium imaging technology. However, due to the anatomical structure and physiological function of the spine, the application and research of in vivo calcium imaging technology for the spinal cord are not very mature at present. The reason is that the spine itself is a movable joint and has a physiological curvature, and mice are very likely to turn their heads to bite off the imaging device fixed on the back when they are free to move. In addition, because the spine swings constantly with movement, it is difficult to fix the focal length of the imaging device, so it is difficult to take stable calcium signals. Therefore, the in vivo calcium signal recording at the level of the spinal cord is still in an imperfect stage. Thus, there is an urgent need for an in vivo calcium imaging device that can take stable calcium signals. SUMMARY

[0004] In order to take stable calcium signals, the present application provides a back-mounted visual in vivo calcium imaging device for mice. The device comprises a device main body, a spine holder and fixing screws. The device main body comprises a track pulley system, four universal wheels and a spine holder base. The track pulley system is composed of two mutually perpendicular metal rods, which are a horizontal track and a vertical track. The bottom end of the vertical track is provided with a microscope fixing platform. The spine holder is connected to the spine holder base of the device main body by four fixing screws and is located below the device main body.

[0005] Optionally, the device main body is a ring-shaped cuboid, and the four corners of the ring-shaped cuboid are respectively provided with the universal wheels. One pulley track is embedded on each of two opposite sides of the ring-shaped cuboid, and the pulley track is connected to the track pulley system. The other two opposite sides extend downward to form the spine holder base.

[0006] Optionally, a sleeve is connected to each end of the transverse track and is nested on the pulley track of the device body, and a sliding sleeve is arranged on the transverse track and is connected to the longitudinal track at the side of the sliding sleeve.

[0007] Optionally, the longitudinal track is connected to the sliding sleeve on the transverse track through a track pulley and can slide longitudinally.

[0008] Optionally, the microscope fixing platform is composed of two vertical parallel clamping pieces, the upper end of the clamping piece is connected to the lower end of the longitudinal track, and a rotating screw is arranged in the middle of the two clamping pieces to change the angle of the clamping piece.

[0009] Optionally, each of the two side vertebral holder bases has two first fixing holes for fixing the vertebral holder.

[0010] Optionally, the vertebral holder is connected by two clamping pieces through a hinge and has second fixing holes on both sides matched with the vertebral holder base.

[0011] The technical scheme provided by the present application has the following beneficial effects:

[0012] It is worth noting that the mouse back-mounted visual in vivo calcium imaging device provided by the present application is a device for visual in vivo calcium imaging of a freely movable mouse. The device skillfully combines a vertebral holder, a track pulley system and a microscope fixing platform to achieve a fixing mode that was difficult to achieve in the past. The device has a simple and reasonable structure. Secondly, the device can hold the spine of a mouse based on a vertebral holder composed of two hinges. The clamping is firm and not easy to relax by using the clamping teeth of the shaft, so that the clamping is more stable. In addition, the mouse can carry the device and move freely in a conscious state, and various studies can be carried out at the spinal cord level in a free motion state, so that the device is flexible to use and effectively solves the problems and difficulties existing in the prior art. In summary, the mouse back-mounted visual in vivo calcium imaging device provided by the present application can fix the spine of a mouse and is not easy to move. The device can be combined with a device body carrying a ultramicroscope. The device has a clever design, a simple and reasonable structure, stable clamping, good flexibility, easy use, strong compatibility and can make the calcium signal recording means more abundant. BRIEF DESCRIPTION OF DRAWINGS

[0013] Fig. 1 is a structural schematic diagram of a mouse back-mounted visual in vivo calcium imaging device provided by an embodiment of the present application;

[0014] Fig. 2 is a three-dimensional structural schematic diagram of a vertebral holder provided by an embodiment of the present application;

[0015] Fig. 3 is a partial enlarged perspective view of a gear part of a vertebral holder provided by the embodiment of the present application;

[0016] The reference signs: 1-device body; 2-vertebral holder; 3-fixing screw; 4-track pulley system; 5-four universal wheels; 6-vertebral holder base; 7-transverse track; 8-longitudinal track; 9-microscope fixing platform; 10-pulley track; 11-sliding sleeve; 12-first fixing hole; 13-clamping piece; 14-hinge; 15-second fixing hole; 16-rotation shaft; 17-gear. DETAILED DESCRIPTION

[0017] In order to make the purpose, technical scheme and advantages of the present application more clear, the following will combine the drawings to make further detailed description of the embodiments of the present application.

[0018] It should be noted that when one component is considered to be "provided on" another component, it can be directly provided on the other component, or a middle component can exist simultaneously; when one component is considered to be "connected" to another component, it can be directly connected to the other component, or a middle component can exist simultaneously.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing the particular embodiments only and is not intended to be limiting of the application.

[0020] Fig. 1 is a structure schematic view of a mouse back portable visual in vivo calcium imaging device provided by the embodiment of the present application; Fig. 2 is a perspective structure schematic view of a vertebral holder provided by the embodiment of the present application; Fig. 3 is a partial enlarged perspective structure schematic view of a gear part of a vertebral holder provided by the embodiment of the present application. Referring to Figs. 1-3 , the device comprises a device body 1, a vertebral holder 2 and a fixing screw 3; the device body 1 comprises a track pulley system 4, four universal wheels 5 and a vertebral holder base 6, the track pulley system 4 is composed of two mutually perpendicular metal rods, which are a transverse track 7 and a longitudinal track 8, and the bottom end of the longitudinal track 8 is provided with a microscope fixing platform 9; the vertebral holder 2 is connected to the vertebral holder base 6 of the device body 1 through four fixing screws 3 and is located in the inner lower part of the device body 1.

[0021] It should be noted that the vertebral clamp 2 is used to clamp the vertebra of the mouse, so as to facilitate the subsequent visualization of the spinal cord dorsal horn neurons and calcium imaging; the fixing screw 3 is used to connect the vertebral clamp 2 and the vertebral clamp base 6 of the device main body 1, so that the connection between the vertebral clamp 2 and the device main body 1 is detachable, which is convenient for storage and use; the track pulley system 4 can move the microscope fixed platform 9 up and down and left and right when the mouse moves, so as to drive the ultramicroscope fixed on the microscope fixed platform 9 to move up and down and left and right, so as to adjust the position relationship between the ultramicroscope and the surface of the mouse spinal cord, and to adjust the observed field of view and focal length in real time, so as to take clear photos and obtain stable calcium signals; the universal wheel 5 is flexible and can support and assist the movement of the mouse when the mouse moves freely, and has simple structure and good mobility.

[0022] Optionally, the device main body 1 is a ring-shaped cuboid, and the universal wheels 5 are arranged at four corners of the ring-shaped cuboid; a pulley track 10 is embedded on two opposite sides of the ring-shaped cuboid, and the pulley track 10 is connected with the track pulley system 4, and the other two opposite sides extend downward to form the vertebral clamp base 6. In this way, the entire device can be integrated into a whole, and the ultramicroscope can be stably carried and imaged during the free movement of the mouse, and the structure is simple and reasonable, firm, the selected material is convenient and durable, light in weight, not easy to rust, and can be used for a long time.

[0023] Optionally, a sleeve is connected to each end of the horizontal track 7 and is nested on the pulley track 10 of the device main body 1, and a sliding sleeve 11 is arranged on the horizontal track 7, and the longitudinal track 8 is connected to the side of the sliding sleeve 11.

[0024] Optionally, the longitudinal track 8 is connected with the sliding sleeve 11 on the horizontal track 7 through a track pulley and can slide longitudinally. It should be noted that this design is flexible and ingenious, and is easy to use. Even if the mouse changes different positions and causes the ultramicroscope to deviate from the field of view or out of focus, the ultramicroscope can be quickly adjusted to the appropriate position through the track pulley system 4.

[0025] Optionally, the microscope fixed platform 9 is composed of two vertically parallel clamping pieces, the upper end of the clamping piece is connected with the lower end of the longitudinal track 8, and a rotating screw is arranged in the middle of the two clamping pieces to change the angle of the clamping piece, so as to clamp the ultramicroscope. It should be noted that the connection between the microscope fixed platform 9 and the lower end of the longitudinal track 8 can be in various modes, which is not limited in the present application, for example, the connection can also be achieved through a screw, and the specific connection mode can be referred to the connection mode between the vertebral clamp 2 and the vertebral clamp base 6 in the present application.

[0026] Optionally, the two sides of the vertebral holder base 6 each have two first fixing holes 12 for connecting the vertebral holder 2. In this way, the vertebral holder 2 can be firmly connected to the device body 1, ensuring that the ultramicroscope is not affected by the movement of the mouse during calcium signal recording.

[0027] Optionally, the vertebral holder 2 is connected by two clamping pieces 13 through a hinge 14, and the two sides of the hinge 14 are provided with second fixing holes 15 matched with the vertebral holder base 6. In addition, the hinge 14 is also provided with a rotating shaft 16, and the rotating shaft 16 is provided with a gear 17, so as to realize clamping the spine of the mouse.

[0028] It should be noted that the gear of the rotating shaft can be used to firmly fix and not easily relax during clamping. In addition, the two sides of the clamping piece 13 are also provided with second fixing holes matched with the vertebral holder base 6, and the vertebral holder 2 can be used to connect the mouse vertebral body and the device body 1 as a whole, so as to make the ultramicroscope closely fit the dorsal horn of the spinal cord, and achieve the purpose of stable observation of the calcium signal of the dorsal horn of the spinal cord.

[0029] The complete use process of the mouse back portable visual in vivo calcium imaging device provided by the application can be: when used, the mouse spine can be first placed between the two hinges 13 of the vertebral holder 2 and fixed firmly, then the four first fixing holes 12 on the vertebral holder base 6 are aligned with the second fixing holes 15 on the vertebral holder 2, then the fixed screw 3 is screwed in to make them tightly combined, then the ultramicroscope is connected with the microscope fixing platform 9, and finally the ultramicroscope is adjusted to a suitable position through the track pulley system 4, so as to observe the activity of the dorsal horn neuron, and thus develop the visual calcium imaging of the dorsal horn neuron.

[0030] It is worth mentioning that the mouse back portable visual in vivo calcium imaging device provided by the application is a device for visual in vivo calcium imaging of a freely movable mouse. The device is combined with a spine holder, a track pulley system and a microscope fixing platform to achieve a fixing mode that is difficult to achieve in the prior art. The device has a simple and reasonable structure. Secondly, the device can hold the spine of the mouse based on the spine holder formed by two pieces of a combined leaf. The pawl of the shaft can be used to firmly fix the mouse during holding and is not easy to relax, so that the holding is more stable. In addition, the mouse can carry the device for autonomous activity in a conscious state, and various studies can be carried out at the spinal cord level in a free motion state, so that the device is flexible to use, and the existing problems and difficulties in the in vivo calcium imaging method of the spinal cord are effectively solved. In summary, the mouse back portable visual in vivo calcium imaging device has the advantages of fixed mouse spine, combination with the device main body carrying the ultramicroscope, ingenious design, simple and reasonable structure, stable holding, good flexibility, convenient use, strong compatibility, and can make the calcium signal recording means more abundant.

[0031] The above only describes the preferred embodiments of the application and is not intended to limit the application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the application shall be included in the protection scope of the application.

Claims

1. A mouse back-carried visual in vivo calcium imaging device, characterized by, The device comprises a device body, a vertebral holder and fixing screws; the device body comprises a track pulley system, four universal wheels and a vertebral holder base, the track pulley system is composed of two mutually perpendicular metal rods, namely a horizontal track and a vertical track, the bottom end of the vertical track is provided with a microscope fixing platform; the vertebral holder is connected with the vertebral holder base of the device body through four fixing screws and is located below and inside the device body; The microscope fixing platform is composed of two vertically parallel clamping pieces, the upper end of the clamping piece is connected with the lower end of the vertical track, and a rotating screw is arranged in the middle of the two clamping pieces to change the angle of the clamping piece; The vertebral holder is connected by two clamping pieces through a hinge, and a second fixing hole matched with the vertebral holder base is arranged on the hinge on both sides, a rotating shaft is further arranged on the hinge, and a gear is arranged on the rotating shaft; The device body is a ring-shaped cuboid, the four corners of the ring-shaped cuboid are respectively provided with the universal wheels; one pulley track is embedded on two opposite sides of the ring-shaped cuboid, the pulley track is connected with the track pulley system, and the other two opposite sides extend downward to the vertebral holder base; The vertebral holder bases on both sides are each provided with two first fixing holes for fixing the vertebral holder, and the vertebral holder connects the vertebral body of the mouse and the device body into one body.

2. The mouse back-carried visualized in vivo calcium imaging apparatus according to claim 1, characterized in that, One sleeve is connected with each end of the horizontal track and is nested on the pulley track of the device body, a sliding sleeve is further arranged on the horizontal track, and the sliding sleeve is connected with the vertical track on the side edge.

3. The mouse back-carried visualized in vivo calcium imaging apparatus according to claim 2, characterized in that, The vertical track is connected with the sliding sleeve on the horizontal track through a track pulley and can slide longitudinally.

Images