CN116058983B - Clamping groove clamping device of single photon fluorescence microscope and clamping groove mounting method thereof - Google Patents
Clamping groove clamping device of single photon fluorescence microscope and clamping groove mounting method thereofInfo
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- CN116058983B CN116058983B CN202211479917.5A CN202211479917A CN116058983B CN 116058983 B CN116058983 B CN 116058983B CN 202211479917 A CN202211479917 A CN 202211479917A CN 116058983 B CN116058983 B CN 116058983B
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- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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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, 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, 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
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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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- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- 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
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
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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
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
- A61B5/0084—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes for introduction into the body, e.g. by catheters
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61B90/00—Instruments, implements or accessories specially adapted for surgery or diagnosis and not covered by any of the groups A61B1/00 - A61B50/00, e.g. for luxation treatment or for protecting wound edges
- A61B90/10—Instruments, implements or accessories specially adapted for surgery or diagnosis and not covered by any of the groups A61B1/00 - A61B50/00, e.g. for luxation treatment or for protecting wound edges for stereotaxic surgery, e.g. frame-based stereotaxis
- A61B90/14—Fixators for body parts, e.g. skull clamps; Constructional details of fixators, e.g. pins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B90/00—Instruments, implements or accessories specially adapted for surgery or diagnosis and not covered by any of the groups A61B1/00 - A61B50/00, e.g. for luxation treatment or for protecting wound edges
- A61B90/20—Surgical microscopes characterised by non-optical aspects
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- G02B21/00—Microscopes
- G02B21/24—Base structure
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements
- G02B7/02—Mountings, adjusting means, or light-tight connections, for optical elements for lenses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
- A61B2503/42—Evaluating a particular growth phase or type of persons or animals for laboratory research
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Abstract
The invention relates to the technical field of neurobiology, in particular to a clamping groove clamping device of a single photon fluorescence microscope and a clamping groove mounting method thereof; the invention comprises a clamping groove clamping body, wherein the lower end of the clamping groove clamping body is in a shape similar to a microscope lens, the upper end of the clamping groove clamping body is provided with a mounting hole used for being connected to the stereotactic instrument, and the lower end of the clamping groove clamping body is used for being inserted into a clamping groove.
Description
Technical Field
The invention relates to the technical field of neurobiology, in particular to a clamping groove clamping device of a single photon fluorescence microscope and a clamping groove mounting method thereof.
Background
The miniature single photon fluorescence imaging is a novel nerve imaging method based on calcium imaging, and the basic principle of the miniature single photon fluorescence imaging is that a virus is used for carrying a calcium ion fluorescence indicator, the virus is used for carrying the calcium ion fluorescence indicator in a target brain region in a targeting way, then the virus infects neurons, the calcium ion indicator is expressed in the neurons, when the neurons are excited, the calcium ion indicator is combined with the calcium ion to emit fluorescence due to the fact that the calcium ion indicator is combined with the calcium ion to emit fluorescence, when the neurons are excited, the calcium ion fluorescence indicator is combined with the calcium ion peak, a fluorescence microscope is used for capturing the explosive fluorescence peak at the time, and the activity and the state of the neurons are analyzed based on fluorescence information.
The miniature single photon fluorescent microscope is mainly divided into two parts, the first part is used for embedding micro lens into virus injecting site, the lens is used as extension of microscope lens to capture fluorescent signal in deep brain tissue, one end of the lens is inserted into brain tissue and the other end is exposed out of skull for docking work with microscope, and the second part is used for installing microscope lens slot on the surface of mouse skull, because the fluorescent signal is recorded by micro microscope in a long time and in regular intermittent process, the microscope number is only one, so that the microscope can be installed on mouse skull only during recording, then the mouse is allowed to move freely, the nerve activity of the mouse under specific action is studied, and the mouse is taken down after recording. This procedure therefore requires a fixed device for inserting a microscope, i.e. a clamping groove, on the surface of the mouse skull.
In the current mainstream implementation, when the clamping groove is installed, the clamping groove needs to be fixed on a microscope head to form a whole with the microscope, the microscope clamping device is used for clamping the microscope, then the clamping device is installed on the stereotactic instrument, the whole formed by the microscope and the clamping groove is accurately adjusted to the proper position on the surface of the skull by the stereotactic instrument, the photocuring resin is smeared on the outer part of the clamping groove and the surface of the skull, after the photocuring resin is solidified, the clamping groove is tightly adhered with the skull, the microscope is removed, and the clamping groove is assembled on the skull of the mouse, so that the process is essentially that the microscope lens is used as a clamp for clamping the clamping groove, and only a microscope clamp is used for clamping the microscope on the market, and the clamping groove is clamped by the microscope clamp for installation. In the follow-up time, when carrying out the formal experiment, insert the microscope in the draw-in groove and screw down, can realize the neural imaging of free movable mouse brain tissue, because micro-microscope is small, and fragile, and because micro single photon fluorescence microscope technique's development stage is limited, the focus in this field at present is micro single photon fluorescence microscope itself, the relevant accessory that matches lacks, after leading to buying the microscope by oneself, there is very big obstacle in later stage use, especially at present do not have appeared the device that is exclusively used in alone centre gripping draw-in groove, only can use the microscope indirectly as the holder of draw-in groove, this has increased micro-microscope damage's risk certainly, and the operation is complicated.
Disclosure of Invention
The invention mainly solves the technical problems of providing a clamping groove clamping device of a single photon fluorescence microscope, which is characterized in that a clamping groove is installed in advance through a clamping groove clamping body, and then the single photon fluorescence microscope is inserted into the clamping groove, so that the process that a microscope lens is indirectly used as a clamp holder to clamp the clamping groove for embedding in the prior art is omitted, the clamping groove embedding process is simplified, the efficiency is improved, and the clamping groove installation method of the single photon fluorescence microscope is also provided.
In order to solve the technical problems, the invention adopts the technical scheme that the clamping groove mounting and clamping device of the single photon fluorescence microscope is provided and is connected with a three-dimensional positioning instrument, wherein the clamping groove mounting and clamping device comprises a clamping groove clamping body, the lower end of the clamping groove clamping body is in a shape similar to a microscope lens, the upper end of the clamping groove clamping body is provided with a mounting hole for being connected with the three-dimensional positioning instrument, and the lower end of the clamping groove clamping body is used for being inserted into a clamping groove.
As an improvement of the invention, the clamping groove clamping body is cylindrical.
As a further improvement of the invention, the stereotactic instrument is provided with a crown-shaped axial arm, the crown-shaped axial arm is provided with a screw hole, and the mounting hole of the clamping groove clamping body is connected with the screw hole of the crown-shaped axial arm through a screw, so that the clamping groove clamping body is fixedly connected to the crown-shaped axial arm.
As a further improvement of the invention, the stereotactic instrument is also provided with a vertical axial arm and a sagittal axial arm.
As a still further development of the invention, the crown axial arm is connected with the vertical axial arm.
As a further improvement of the invention, the cylindrical cavity below the bottom surface of the clamping groove carries a micro lens.
A clamping groove installation method of a single photon fluorescence microscope comprises the following steps:
Step S1, fixing the head of a mouse on a stereotactic instrument, cutting off the scalp of the mouse, exposing the skull of the mouse, establishing a coordinate system on the skull through a bone seam on the surface of the skull according to a neurobiological method, finding a target site, and drilling holes in the target site by using a skull drill to penetrate the skull;
S2, inserting a clamping groove into the lower end of the clamping groove clamping body, and connecting the upper end of the clamping groove clamping body to the three-dimensional positioning instrument through a screw;
s3, operating the stereotactic instrument, and moving the clamping groove clamping body to the upper part of the head of the mouse through the crown-shaped axial arm of the stereotactic instrument, so that the clamping groove connected to the lower end of the clamping groove clamping body is positioned at a target site of the head of the mouse;
s4, operating the stereotactic instrument, and driving the clamping groove clamping body to move downwards through the vertical axial arm and the coronal axial arm of the stereotactic instrument so that the micro lens in the clamping groove is inserted into brain tissue;
and S5, when the micro lens in the clamping groove is inserted into the target depth, using dental cement to adhere the outer wall of the clamping groove to the surface of the skull, and after the dental cement is solidified, pulling out the clamping groove clamping body, so that the clamping groove is firmly adhered to the surface of the skull of the mouse, and at the moment, the installation process of the clamping groove is completed.
Compared with the prior art, the clamping groove clamping device has the advantages that the clamping groove is installed in advance through the clamping groove clamping body, and then the single photon fluorescence microscope is inserted into the clamping groove, so that the process that the clamping groove is clamped by the microscope lens indirectly serving as a clamp holder for embedding in the prior art is omitted, the clamping groove embedding process is simplified, and the efficiency is improved.
Drawings
FIG. 1 is a schematic illustration of an application of the present invention;
FIG. 2 is a schematic diagram of a clamping slot clamping body according to the present invention;
FIG. 3 is a schematic diagram of a card slot according to the present invention;
reference numerals are 1-screw hole, 2-vertical axial arm, 3-sagittal axial arm, 4-coronal axial arm, 5-set screw, 6-clamping groove clamping body and 7-mouse.
Detailed Description
Referring to fig. 1 to 3, a clamping device for a single photon fluorescence microscope according to the present invention is connected to a stereotactic apparatus, and includes a clamping body 6, wherein the lower end of the clamping body 6 is shaped like a microscope lens, the upper end of the clamping body 6 is provided with a mounting hole for connecting to the stereotactic apparatus, and the lower end of the clamping body 6 is used for inserting a clamping groove.
In the invention, the clamping groove is arranged in advance through the clamping groove clamping body 6, and then the single photon fluorescence microscope is inserted into the clamping groove, so that the process of clamping the clamping groove by indirectly taking the microscope lens as a clamp in the prior scheme is omitted, the clamping groove embedding process is simplified, and the efficiency is improved.
In the present invention, the clamping groove clamping body 6 is cylindrical.
In the invention, a crown axial arm 4 is arranged on the stereotactic instrument, a screw hole 1 is arranged on the crown axial arm 4, a mounting hole of a clamping groove clamping body 6 is connected with the screw hole 1 of the crown axial arm 4 through screws, so that the clamping groove clamping body 6 is fixedly connected to the crown axial arm 4, a vertical axial arm 2 for lifting in the vertical direction and a sagittal axial arm 3 for moving forwards and backwards are also arranged on the stereotactic instrument, and the crown axial arm 4 is connected with the vertical axial arm 2.
In the invention, the cylindrical cavity below the bottom surface of the clamping groove carries the micro lens.
Specifically, according to the size of the miniature single photon microscope lens, a 3D printing technology is used to design a clamping groove clamping body 6 capable of stably clamping the clamping groove of the miniature microscope lens, the clamping groove clamping body 6 is integrally shaped into a cylinder, the upper end of the clamping groove clamping body is connected with a screw hole 1 on a crown-shaped axial arm 4 of the stereotactic instrument through a screw, and the lower end of the clamping groove clamping body is designed into the shape of the microscope lens.
The invention also provides a clamping groove mounting method of the single photon fluorescence microscope, which comprises the following steps:
Step S1, fixing the head of a mouse on a stereotactic instrument, cutting off the scalp of the mouse, exposing the skull of the mouse, establishing a coordinate system on the skull through a bone seam on the surface of the skull according to a neurobiological method, finding a target site, and drilling holes in the target site by using a skull drill to penetrate the skull;
s2, inserting a clamping groove into the lower end of the clamping groove clamping body 6, and connecting the upper end of the clamping groove clamping body 6 to the three-dimensional positioning instrument through a screw;
s3, operating the stereotactic instrument, and moving the clamping groove clamping body to the upper part of the head of the mouse through the crown-shaped axial arm 4 of the stereotactic instrument, so that the clamping groove connected to the lower end of the clamping groove clamping body 6 is positioned at a target site of the head of the mouse;
S4, operating the stereotactic instrument, and driving the clamping groove clamping body 6 to move downwards through the vertical axial arm 2 and the coronal axial arm 4 of the stereotactic instrument, so that the micro lens in the clamping groove is inserted into brain tissue;
and S5, when the micro lens in the clamping groove is inserted into the target depth, using dental cement to adhere the outer wall of the clamping groove to the surface of the skull, and after the dental cement is solidified, pulling out the clamping groove clamping body, so that the clamping groove is firmly adhered to the surface of the skull of the mouse, and at the moment, the installation process of the clamping groove is completed.
The method comprises the steps of fixing the head of a mouse on a positioning instrument, cutting off the scalp to expose the skull, establishing a coordinate system on the skull through a bone seam on the surface of the skull according to a neurobiological method, finding a target site, drilling a hole on the surface of the skull by using a skull drill, inserting the lower end of a clamping groove clamping body 6 into a clamping groove, connecting the upper end of the clamping groove clamping body with the stereotactic instrument through a screw, fixing the whole clamping groove clamping body 6 on the stereotactic instrument, operating the stereotactic instrument at the moment, adjusting the clamping groove to be above the drilled hole, carrying a micro lens in a cylindrical official cavity below the bottom of the clamping groove, then operating a clamping groove clamp 6 downwards by using a vertical axial arm 2 of the stereotactic instrument, inserting the micro lens carried by the lower part of the clamping groove into brain tissue, adhering the outer wall of the clamping groove to the surface of the skull when the clamping groove is inserted into the target depth, loosening a screw on the clamping groove after the dental cement is solidified, and firmly adhering the clamping groove on the surface of the forehead mouse at the moment, thus completing the installation process of the clamping groove. In the formal experiment, the lens of the micro microscope is inserted into the clamping groove to record the calcium ion fluorescence signal.
The invention simulates the lens of a microscope to manufacture a clamping groove clamp, clamps the microscope by the microscope clamp before, then installs the clamping groove on the microscope, then uses the whole to embed, reduces the two processes into a process of directly using the clamp to clamp the clamping groove for embedding.
The invention solves the problem of difficult clamping of the clamping groove of the microscope when the micro single photon fluorescence microscope is installed on the surface of the skull of the mouse, changes the prior clamping groove installation process, only has the microscope clamp on the market but does not have a special clamping groove clamp, and needs to install the clamping groove on the microscope by means of the micro microscope, then uses the microscope clamp to install the whole, and then removes the microscope after the installation is completed.
The foregoing description is only of embodiments of the present invention, and is not intended to limit the scope of the invention, and all equivalent structures or equivalent processes using the descriptions and the drawings of the present invention or directly or indirectly applied to other related technical fields are included in the scope of the present invention.
Claims (6)
1. The clamping groove mounting and clamping device of the single photon fluorescence microscope is connected with a stereotactic instrument and is characterized by comprising a clamping groove clamping body, wherein the lower end of the clamping groove clamping body is in a shape similar to a microscope lens and is used for being directly inserted into a clamping groove to replace the microscope lens in the clamping groove mounting process, the upper end of the clamping groove clamping body is provided with a mounting hole used for being connected with the stereotactic instrument, and a vertical axial arm is further arranged on the stereotactic instrument.
2. The clamping groove mounting and clamping device of a single photon fluorescence microscope according to claim 1, wherein the clamping groove clamping body is cylindrical.
3. The clamping groove mounting and clamping device of a single photon fluorescence microscope according to claim 2, wherein a crown-shaped axial arm is arranged on the stereotactic instrument, a screw hole is arranged on the crown-shaped axial arm, and the mounting hole of the clamping groove clamping body is connected with the screw hole of the crown-shaped axial arm through a screw, so that the clamping groove clamping body is fixedly connected to the crown-shaped axial arm.
4. A clamping groove mounting and clamping device for a single photon fluorescence microscope as in claim 3 wherein said coronal axial arms are connected to said vertical axial arms.
5. The clamping groove mounting and clamping device of a single photon fluorescence microscope according to claim 4, wherein a cylindrical cavity below the bottom surface of the clamping groove carries a micro lens.
6. The clamping groove installation method of the single photon fluorescence microscope is characterized by comprising the following steps of:
Step S1, fixing the head of a mouse on a stereotactic instrument, cutting off the scalp of the mouse to expose the skull, establishing a coordinate system on the skull through a bone seam on the surface of the skull according to a neurobiological method, finding a target site, and drilling holes in the target site by using a skull drill to penetrate the skull;
S2, inserting a clamping groove into the lower end of the clamping groove clamping body, and connecting the upper end of the clamping groove clamping body to the three-dimensional positioning instrument through a screw;
s3, operating the stereotactic instrument, and moving the clamping groove clamping body to the upper part of the head of the mouse through the crown-shaped axial arm of the stereotactic instrument, so that the clamping groove connected to the lower end of the clamping groove clamping body is positioned at a target site of the head of the mouse;
s4, operating the stereotactic instrument, and driving the clamping groove clamping body to move downwards through the vertical axial arm and the coronal axial arm of the stereotactic instrument so that the micro lens in the clamping groove is inserted into brain tissue;
and S5, when the micro lens in the clamping groove is inserted into the target depth, using dental cement to adhere the outer wall of the clamping groove to the surface of the skull, and after the dental cement is solidified, pulling out the clamping groove clamping body, so that the clamping groove is firmly adhered to the surface of the skull of the mouse, and at the moment, the installation process of the clamping groove is completed.
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| CN202211479917.5A CN116058983B (en) | 2022-11-24 | 2022-11-24 | Clamping groove clamping device of single photon fluorescence microscope and clamping groove mounting method thereof |
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| CN202211479917.5A CN116058983B (en) | 2022-11-24 | 2022-11-24 | Clamping groove clamping device of single photon fluorescence microscope and clamping groove mounting method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN211674365U (en) * | 2019-12-13 | 2020-10-16 | 中国科学院深圳先进技术研究院 | Clamp holder |
| CN114287881A (en) * | 2021-12-11 | 2022-04-08 | 中国科学院深圳先进技术研究院 | Micro single-photon fluorescence microscope implantation device and implantation method |
| CN219021600U (en) * | 2022-11-24 | 2023-05-16 | 中国科学院深圳先进技术研究院 | A card slot clamping device for a single-photon fluorescence microscope |
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| CN109172025A (en) * | 2018-10-09 | 2019-01-11 | 中国科学技术大学 | Animal brain surgical apparatus and its control device, control system |
| CN110946660B (en) * | 2019-12-30 | 2021-05-07 | 中国科学技术大学 | Adjustable double-hole ferrule pin clamping device and stereotaxic apparatus including the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN211674365U (en) * | 2019-12-13 | 2020-10-16 | 中国科学院深圳先进技术研究院 | Clamp holder |
| CN114287881A (en) * | 2021-12-11 | 2022-04-08 | 中国科学院深圳先进技术研究院 | Micro single-photon fluorescence microscope implantation device and implantation method |
| CN219021600U (en) * | 2022-11-24 | 2023-05-16 | 中国科学院深圳先进技术研究院 | A card slot clamping device for a single-photon fluorescence microscope |
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