阅读说明：本技术 一种可进行光遗传刺激的微透析探针 (Microdialysis probe capable of carrying out optogenetic stimulation ) 是由 陈嘉志 陈心语 李楠翔 黄曙杰 文焕涛 王军 张旺明 于 2019-08-06 设计创作，主要内容包括：本发明公开了一种可进行光遗传刺激的微透析探针,其包括探针壳体、限位块、光纤连接头,探针壳体头部连接与限位块,光纤连接头与限位块结合,此三个结构构成本发明的主体结构,本发明将光纤模块和透析模块相结合,实现了同步进行光遗传刺激和微量渗析这两种操作,能够实时同步地检测光波激活特定神经元后带来的分子水平变化；只需要在进行光刺激和微透析操作时将其插入目标脑区,这样可以避免因长期放置光纤与脑内的传统方法造成的神经元损伤和组织反应,这样有利于长期跟踪实验。此发明用于生物医疗器械领域。(The invention discloses a microdialysis probe capable of carrying out optogenetic stimulation, which comprises a probe shell, a limiting block and an optical fiber connector, wherein the head part of the probe shell is connected with the limiting block, the optical fiber connector is combined with the limiting block, and the three structures form a main structure of the microdialysis probe; only need insert it in target brain area when carrying out light stimulation and microdialysis operation, can avoid because of long-term placing optic fibre and the neuron injury and the tissue reaction that traditional method in the brain caused, so be favorable to long-term tracking experiment. The invention is used in the field of biomedical devices.)

1. a microdialysis probe capable of optogenetic stimulation comprising: the dialysis system comprises an optical fiber module and a dialysis module, wherein the optical fiber module comprises an optical fiber connector (1) and a quartz optical fiber (2);

The dialysis module comprises a probe shell (3), two cavities are arranged in the probe shell (3), and each cavity comprises a perfusion liquid passing cavity (31) positioned on the inner side and a dialysis liquid passing cavity (32) positioned on the outer side; a horn-shaped first opening (33) is designed at the bottom end of the probe shell (3), a dialysis membrane (4) is mounted on the conical surface of the first opening (33), and the head of the probe shell (3) is connected to a limiting block (5); the optical fiber connector (1) is combined with the limiting block (5); the quartz optical fiber (2) is arranged in the optical fiber connector (11), the limiting block (5) and the probe shell (3);

The perfusion liquid inlet at the upper end of the perfusion liquid passing cavity (31) is connected with a perfusion liquid input pipe (34), the bottom end of the perfusion liquid passing cavity (31) is provided with a second opening (35), and the dialysis liquid passing cavity (32) is communicated with the perfusion liquid passing cavity (31) through the second opening (35); the dialysate is connected to a dialysate outlet line (36) via an inlet port at the upper end of the chamber (32).

2. The microdialysis probe according to claim 1 which is capable of optogenetically stimulating, characterized by: an isolation layer covers the outer side of the quartz optical fiber (2) in the probe shell (3).

3. the microdialysis probe according to claim 2 which is capable of optogenetically stimulating, characterized by: the bottom end of the quartz optical fiber (2) and the top end of the first opening (33) of the probe shell (3) are positioned on the same plane.

4. The microdialysis probe according to claim 3 which is capable of optogenetically stimulating, characterized by: the quartz optical fiber (2) is located in the center of the optical fiber connector (1), the limiting block (5) and the probe shell (3).

5. The microdialysis probe according to claim 1 which is capable of optogenetically stimulating, characterized by: the perfusion fluid input tube (34) and the dialysate output tube (36) are buried in the optical fiber connector (1), and the openings of the perfusion fluid input tube (34) and the dialysate output tube (36) are located at the same side of the optical fiber connector (1).

6. The microdialysis probe according to claim 1 which is capable of optogenetically stimulating, characterized by: the dialysis membrane (4) is an annular dialysis membrane mounted on the first opening (33).

Technical Field

The invention relates to the field of biomedical equipment, in particular to a microdialysis probe capable of carrying out optogenetic stimulation.

Background

Dialysis is a common technique in the field of life science, and the main principle is to separate small molecules from biomacromolecules by using a semipermeable membrane, and dialyze and exchange perfusion fluid and extracellular fluid for low-molecular-weight substances to be taken out and measured, so as to collect and analyze the substances. The optogenetic technology is the leading-edge technology of current research in the field of neuroscience, and the basic principle is that a gene manipulation technology is utilized to transfect specific genes capable of coding light-sensitive proteins, such as ChR2, NaHR and the like, to specific types of cells to express special ion channels. Under the action of light-sensitive protein, the ion channel can produce selectivity for the passing of cation or anion under the stimulation of different wavelengths of light, so that the membrane potential on two sides of cell membrane can be changed, and the protein expressed by said gene can be activated or inhibited by light signal, so that the specific type of neuron can be activated or inhibited. Further, after activating or inhibiting a specific type of neuron in a certain brain region using optogenetic techniques or after obtaining a behavioral improvement, it is often necessary to further detect biochemical information, or changes in electrophysiological information, in the brain region and downstream brain regions. This allows one to explore whether the level of release of a small molecule (e.g., dopamine neurotransmitter) is mediated by a certain type of neuron, or whether the behavioral improvement in a disease is mediated by an altered level of a small molecule. However, the prior art can only realize the light stimulation in the upstream brain area and the collection of biochemical information in the downstream brain area, but cannot perform real-time synchronous biochemical recording (for example, collecting certain small molecular substances through microdialysis) in a stimulation target, which is not convincing to explain the pathophysiological change of a light stimulation target area.

The quartz optical fiber used in the laboratory at present is often required to be placed in the brain of an experimental animal for a long time. Due to the mechanical property of the quartz optical fiber, certain extrusion damage can be generated to fragile brain tissues, and the influence on the experimental effect is large. At present, no equipment report about the combination of a preposed optical fiber of a light stimulation device and micro-dialysis to achieve synchronous detection exists, and the device capable of synchronously detecting the concentration of light stimulation and small molecular substances has scientific research requirements.

Disclosure of Invention

the invention aims to provide a microdialysis probe which has the functions of photostimulation and dialysis of small molecular substances in a target region and can carry out optogenetic stimulation.

The technical scheme adopted by the invention is as follows:

A microdialysis probe capable of carrying out optogenetic stimulation comprises an optical fiber module and a dialysis module, wherein the optical fiber module comprises an optical fiber connector and a quartz optical fiber. The micro-dialysis probe integrated combination of the optical fiber module and the dialysis module only needs to be inserted into a target brain area when light stimulation and micro-dialysis operation are carried out, so that neuron damage and tissue reaction caused by long-term placement of the optical fiber in the brain are avoided, and long-term tracking experiments are facilitated.

the dialysis module comprises a probe shell, two cavities are arranged in the probe shell, and each cavity comprises a perfusion liquid passing cavity positioned on the inner side and a dialysis liquid passing cavity positioned on the outer side; the bottom design of probe casing has the first opening of loudspeaker form, install the dialysis membrane on the conical surface of first opening, the head of probe casing is connected on a stopper, and the stopper plays the effect of restriction probe slip in the sheathed tube of implanting in the experiment. The optical fiber connector is combined with the limiting block; the quartz optical fiber is arranged in the optical fiber connector, the limiting block and the probe shell.

The upper end input port of the perfusate passing cavity is connected with a perfusate input pipe, the bottom end of the perfusate passing cavity is provided with a second opening, and the dialyzate passing cavity is communicated with the perfusate passing cavity through the second opening; the dialysate is connected with a dialysate output pipe through an input port at the upper end of the chamber. The dialysate output tube is used for discharging dialysate, and the dialysate is further used for performing high performance liquid chromatography.

Further as an improvement of the technical scheme of the invention, an isolation layer covers the outer side of the quartz optical fiber in the probe shell, so that the quartz optical fiber can be prevented from being directly contacted with the perfusion liquid.

As a further improvement of the technical solution of the present invention, the bottom end of the silica optical fiber and the top end of the first opening of the probe housing are located on the same plane, and the arrangement is mainly for releasing light waves with a certain wavelength.

As further improvement of the technical scheme of the invention, the quartz optical fiber is positioned in the centers of the optical fiber connector, the limiting block and the probe shell.

As a further improvement of the technical scheme of the present invention, the perfusion fluid input tube and the dialysate output tube are both embedded in the optical fiber connector, and the openings of the perfusion fluid input tube and the dialysate output tube are located at the same side of the optical fiber connector.

As a further improvement of the technical solution of the present invention, the dialysis membrane is a ring-shaped dialysis membrane installed on the first opening.

the invention has the beneficial effects that: the microdialysis probe capable of carrying out optogenetic stimulation comprises a probe shell, a limiting block and an optical fiber connector, wherein the head of the probe shell is connected with the limiting block, the optical fiber connector is combined with the limiting block, and the three structures form a main structure of the microdialysis probe; the optical fiber module and the dialysis module are combined, so that the two operations of optogenetic stimulation and microdialysis are synchronously performed, and the molecular level change caused by the activation of specific neurons by light waves can be synchronously detected in real time; the bottom of the probe shell is provided with a horn-shaped first opening, on one hand, the first opening is just corresponding to the radiation angle of light waves, so that biochemical signals of neurons stimulated by light can be collected to the maximum degree, on the other hand, a dialysis membrane is arranged on the horn-shaped first opening, the dialysis membrane can dialyze molecules of extracellular fluid of the neurons in the region stimulated by the light to the maximum degree, and synchronous molecular level change brought by the light stimulation can be collected and recorded to the maximum degree; meanwhile, the optical fiber module and the dialysis module are integrated, and only the optical fiber module is required to be inserted into a target brain area when optical stimulation and microdialysis are carried out, so that neuron damage and tissue reaction caused by the traditional method of placing the optical fiber and the brain for a long time can be avoided, and long-term tracking experiments are facilitated.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

FIG. 1 is a front view of an embodiment of the present invention;

FIG. 2 is a top view of an embodiment of the present invention;

FIG. 3 is a bottom view of an embodiment of the present invention;

FIG. 4 is a front view of a probe housing of an embodiment of the invention;

FIG. 5 is a state diagram of an embodiment of the present invention;

FIG. 6 is a diagram illustrating the state of the embodiment of the present invention in the light stimulation state.

Detailed Description

Referring to fig. 1 to 6, the present invention is a microdialysis probe capable of performing optogenetic stimulation, which comprises an optical fiber module and a dialysis module, wherein the optical fiber module comprises an optical fiber connector 1 and a quartz optical fiber 2. The optical fiber module and the dialysis module are combined, so that the two operations of optogenetic stimulation and microdialysis are synchronously performed, and the molecular level change caused by the activation of specific neurons by light waves can be synchronously detected in real time.

The dialysis module comprises a probe housing 3, wherein two cavities are arranged in the probe housing 3, and each cavity comprises a perfusion liquid passing cavity 31 positioned on the inner side and a dialysis liquid passing cavity 32 positioned on the outer side. The perfusion liquid inlet at the upper end of the perfusion liquid passing cavity 31 is connected with a perfusion liquid inlet pipe 34, the bottom end of the perfusion liquid passing cavity 31 is provided with a second opening 35, and the dialysate passing cavity 32 is communicated with the perfusion liquid passing cavity 31 through the second opening 35; the dialysate is connected to a dialysate output line 36 through an upper input port of the chamber 32. The second opening 35 is a ring-shaped opening, in the experiment, the perfusate enters the perfusate passing chamber 31 from the perfusate input tube 34, after the dialysis of the molecular substances in the dialysis membrane 4 area is completed, the perfusate passes through the second opening 35 and enters the dialyzate passing chamber 32, and then the dialyzate is output through the dialyzate output tube 36, and the output dialyzate can be subjected to biochemical analysis such as high performance liquid chromatography. In a preferred embodiment of the present invention, the quartz optical fiber 2 inside the probe housing 3 is covered with an isolation layer. The isolation layer is mainly used for avoiding direct contact between the perfusion liquid and the quartz optical fiber 2.

The bottom of the probe shell 3 is provided with a horn-shaped first opening 33, the first opening 33 is designed to correspond to the radiation angle of light waves, so that biochemical signals of neurons stimulated by light can be collected to the maximum degree, and the first opening 33 is provided with a dialysis membrane 4, so that molecules of extracellular fluid of the neurons in the area can be dialyzed to the maximum degree after being stimulated by the light by the dialysis membrane 4, and synchronous molecular level changes caused by the light stimulation can be collected and recorded to the maximum degree.

In this embodiment, the bottom end of the quartz fiber 2 is preferably disposed on the same plane as the top end of the first opening 33 of the probe housing 3 for releasing light with a certain wavelength, such as blue light with a wavelength of 473nm and yellow light with a wavelength of 593 nm.

The bottom design of probe casing 3 has the first opening 33 of loudspeaker form, install dialysis membrane 4 on the conical surface of first opening 33, the head of probe casing 3 is connected on a stopper 5, quartz fiber 2 is located optical fiber connector 1, stopper 5 and probe casing 3's central authorities. The optical fiber connector 1 is combined with the limiting block 5; and the quartz optical fiber 2 is arranged in the optical fiber connector 1, the limiting block 5 and the probe shell 3. The probe housing 3, the optical fiber connector 1 and the limiting block 5 form a main body structure of the present invention, as shown in fig. 4 and 5, the bottom end of the probe housing 3 is designed into a first opening 33 in a horn shape, and a dialysis membrane 4 is laid on the circular periphery of the first opening 33. Further, the dialysis membrane 4 is a ring-shaped dialysis membrane mounted on the first opening 33. The dialysis membrane 4 is mainly used for dialyzing molecules with a certain molecular weight of extracellular fluid of neurons in a target brain region.

In a specific embodiment, a connecting cap 6 is installed on the top of the optical fiber connector 1, a jumper 7 is connected to the connecting cap 6, one end of the jumper 7 is connected to the input end of the quartz optical fiber 2, and the other end of the jumper 7 is connected to the optogenetic instrument 8. After the optical fiber connector 1 is connected to the connection cap 6, the light wave with a certain wavelength excited by the optogenetic instrument 8 enters the quartz fiber 2 through the jumper 7 and is released at the bottom end of the quartz fiber 2, as shown in fig. 6, the light wave with a certain wavelength and intensity is released in a radiation mode, so that the neurons in the horn-shaped first opening 33 are controlled, yellow light with a wavelength of 593nm or blue light with an activation wavelength of 473nm are inhibited, and meanwhile, real-time molecular level changes brought by the controlled neurons are collected by the dialysis probe and are reflected on a next-stage molecular level detection instrument.

in a preferred embodiment of the present invention, the perfusion fluid input tube 34 and the dialysate output tube 36 are both embedded in the optical fiber connector 1, and the perfusion fluid input tube 34 and the dialysate output tube 36 are open on the same side of the optical fiber connector 1.

the optical fiber module and the dialysis module are integrated, and only the optical fiber module is inserted into a target brain area during light stimulation and microdialysis operation, so that neuron damage and tissue reaction caused by long-term placement of the optical fiber and a traditional method in the brain can be avoided, and long-term tracking experiments are facilitated.

The invention is not limited to the above embodiments, and those skilled in the art can make equivalent modifications or substitutions without departing from the spirit of the invention, and such equivalent modifications or substitutions are included in the scope defined by the claims of the present application.