CN111437514A - Optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection and preparation method thereof - Google Patents
Optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection and preparation method thereof Download PDFInfo
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- CN111437514A CN111437514A CN202010246172.2A CN202010246172A CN111437514A CN 111437514 A CN111437514 A CN 111437514A CN 202010246172 A CN202010246172 A CN 202010246172A CN 111437514 A CN111437514 A CN 111437514A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
- A61N5/00—Radiation therapy
- A61N5/06—Radiation therapy using light
- A61N5/0601—Apparatus for use inside the body
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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
- A61B5/24—Detecting, measuring or recording bioelectric or biomagnetic signals of the body or parts thereof
- A61B5/25—Bioelectric electrodes therefor
- A61B5/279—Bioelectric electrodes therefor specially adapted for particular uses
- A61B5/291—Bioelectric electrodes therefor specially adapted for particular uses for electroencephalography [EEG]
Abstract
The invention provides an optical fiber probe for simultaneously realizing deep brain light stimulation and brain electrical detection, which can synchronously realize the fixed-point accurate acquisition of light stimulation and brain electrical signals through one optical fiber. Comprises an optical fiber 1, which is used for introducing laser under certain conditions and inserting a part of the optical fiber into the brain of a living body to realize light stimulation; the outside of the optical fiber 1 is coated with a conductive film 2 and an insulating film 3; a detection area which is not covered by the insulating film 3 is reserved at a specific position on the optical fiber conducting film 2 inserted into the brain of the organism and is used for realizing the electroencephalogram detection of the specific position; the optical fiber left outside the brain is inserted in the ceramic ferrule 4, the optical fiber on the top of the ceramic ferrule 4 is provided with a boundary region which does not cover the insulating film 3, and the detection lead 6 is connected with the conductive film through the conductive adhesive 5 on the boundary region to realize the export of the brain electrical signal; the ceramic ferrule 4 is externally sleeved with a ceramic sleeve 7. The invention also provides a preparation method of the optical fiber probe, which has the advantages of simple manufacturing process, small structure and excellent biocompatibility.
Description
Technical Field
The invention belongs to the field of optogenetics, and particularly relates to an optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection and a preparation method thereof.
Background
Optogenetics is a technology developed in recent years by combining optics with genetics in organisms, introducing modified viruses into brain tissues of a subject organism, and stimulating channel proteins of specific cells by introducing laser light of different wavelengths, frequencies, and pulses into optical fibers implanted in the brain tissues of the organism, thereby regulating the behavior of the organism. In recent years, the design of multifunctional optical fibers has become a hotspot of research in this field.
The defects of the prior art are as follows: the traditional optical fiber only has the function of transmitting light, but does not have the function of detecting electroencephalogram signals. Therefore, when the relation between the light stimulation and the electroencephalogram signals of the animals needs to be researched, the light stimulation equipment and the electroencephalogram detection equipment need to be implanted into the animals at the same time, so that the brain of the organisms can be greatly damaged, the immunoreaction can be more easily caused, and the detection fails.
Disclosure of Invention
In order to fill the blank of the prior art, the invention aims to provide the optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection, can synchronously realize fixed-point accurate detection and recording of light stimulation and electroencephalogram signals through one optical fiber, and simultaneously realize effective light stimulation in a biological brain and acquisition of electroencephalogram signals at specific positions; the invention also provides a preparation method of the optical fiber probe, which has the advantages of simple manufacturing process, small structure and excellent biocompatibility.
The technical scheme adopted by the invention for solving the technical problems is as follows:
an optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection is characterized by comprising an optical fiber 1, wherein the optical fiber 1 is used for introducing laser under certain conditions, and inserting one part of the optical fiber into the brain of a living body to realize light stimulation; the outside of the optical fiber 1 is coated with a conductive film 2, and the outside of the conductive film 2 is coated with an insulating film 3; a detection area which is not covered by the insulating film 3 is reserved at a specific position on the optical fiber conducting film 2 inserted into the brain of the organism and is used for directly contacting nerve cells and tissue fluid to realize the electroencephalogram detection at the specific position; the optical fiber left outside the brain is inserted in the ceramic ferrule 4, the optical fiber on the top of the ceramic ferrule 4 is provided with a boundary area which does not cover the insulating film 3, and the boundary area is used for connecting the detection lead 6 with the conductive film through the conductive adhesive 5 so as to realize the derivation of the electroencephalogram signal; the ceramic ferrule 4 is externally sleeved with a ceramic sleeve 7.
The conductive film 2 is made of materials with impedance characteristics and biocompatibility, and comprises titanium, platinum and gold.
The conductive film 2 is preferably made of titanium.
The insulating film 3 is made of Parylene (Parylene).
The detection zone has an axial length of at least 25 microns to ensure that the detection zone can accurately correspond to a single or a few nerve cells or nerve fibers.
The detection lead 6 is a copper wire with the diameter of 0.5mm-1 mm; the interface region has an axial length of at least 2 mm.
The ceramic sleeve 7 is matched with the ceramic ferrule 4, the inner diameter of the ceramic ferrule is matched with the outer diameter of the coated optical fiber,
the diameter of the optical fiber 1 is less than 200 microns, and the requirement of not damaging biological tissues is met.
A method for preparing an optical fiber probe for simultaneously realizing deep brain light stimulation and brain electrical detection is characterized by comprising the following steps:
s1 plating conductive film on optical fiber
Preparing an optical fiber with proper length, uniformly plating a conductive material on the optical fiber by utilizing a vacuum coating technology, coating a conductive film outside the optical fiber, wherein the impedance of the conductive film meets the following requirements: under the condition of 1kHz, the sum of the impedance of the copper wire and the impedance of the copper wire is less than 500k omega; the impedance characteristic of the brain electrical signals can be used for recording the brain electrical signals;
s2 coating the optical fiber with an insulating film
Uniformly plating Parylene (Parylene) outside the conductive film except the detection area and the boundary area by using a vacuum coating technology; the thicknesses of the conductive film and the insulating film meet the requirement that the diameter of the optical fiber is matched with the inner diameter of the ceramic ferrule after film coating;
s3 connection of optical fiber and ceramic ferrule
Inserting the optical fiber left outside the brain into the ceramic ferrule, wherein the end part of the optical fiber is aligned with the ceramic ferrule; sleeving the ceramic sleeve outside the ceramic ferrule and aligning with the ceramic ferrule; therefore, the convenience and the stability of the optical fiber device in the using process are ensured;
s4 connection of copper wire to conductive film
Connecting copper wires on the boundary area; clamping the coated optical fiber by using an optical fiber clamp, and connecting the copper wire and the conductive film under a microscope; the two copper wires are in central symmetry with the axis of the optical fiber, and the conductive adhesive completely covers the contact point of the copper wires and the conductive film, so that the electroencephalogram signals in a living body can be led out through the copper wires.
In the step S2, the insulating film plating method includes two methods:
the first method is a shielding method: shielding a detection area and a boundary area which do not need to be plated with an insulating film by using a proper material, cleaning and pretreating the optical fiber plated with a conductive film, conveying the treated optical fiber into a vacuum furnace, sublimating, cracking and depositing an insulating film raw material on the surface of a test piece to form a layer of compact high-molecular insulating film;
the second method is a photoetching method: after cleaning the optical fiber, spin-coating photoresist as a mask material on the conducting film for the detection area and the boundary area which do not need to be plated with the insulating film by a chemical vapor deposition method, and then carrying out photoetching to obtain a required sample, wherein the precision of the method is usually determined by the resolution of a photoetching link.
The invention has the technical effects that:
compared with the prior art, the invention has the advantages that:
(1) one optical fiber can synchronously realize laser stimulation and electroencephalogram signal acquisition, and the blank of the field is filled.
(2) The position of the electroencephalogram detection area is variable. For the part, in the brain, of the optical fiber, a detection area with the axial length of at least 25 micrometers can be reserved at any position along the axial direction, namely the detection area can be located at any position of the implanted part, and accurate detection and recording of electroencephalogram signals at specific positions can be achieved.
(3) The technology is simple and easy to realize.
(4) The optical fiber structure is small and exquisite, has good biocompatibility, avoids causing the injury to biological tissue.
Drawings
FIG. 1 is an overall structure diagram of an optical fiber according to the present invention after two coating processes (the detection zone is located at the foremost end);
FIG. 2 is a partial block diagram of the optical fiber detection zone after two coating passes in accordance with the present invention (the detection zone is located at the foremost end);
FIG. 3 is an overall structure diagram of the optical fiber after two coating processes (the detection zone is located in the middle) according to the present invention;
FIG. 4 is a block diagram of the coated optical fiber of the present invention after the ferrule and the ferrule are attached;
FIG. 5 is a diagram showing the relative positions of copper wires, conductive adhesive and optical fibers according to the present invention;
FIG. 6 is a view showing the construction of the overall apparatus of the present invention;
the reference numbers are listed below: 1-an optical fiber; 2-conductive titanium film; 3-an insulating film; 4-ceramic ferrule; 5-conductive adhesive; 6-copper wire; 7-ceramic bushing.
Detailed Description
The invention is further described with reference to the following figures and detailed description.
As shown in fig. 1-6, an optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection comprises an optical fiber 1, which is used for introducing laser under certain conditions, and inserting a part of the optical fiber into the brain of a living body to realize light stimulation; the outside of the optical fiber 1 is coated with a conductive film 2, and the outside of the conductive film 2 is coated with an insulating film 3; a detection area which is not covered by the insulating film 3 is reserved at a specific position on the optical fiber conducting film 2 inserted into the brain of the organism and is used for directly contacting nerve cells and tissue fluid to realize the electroencephalogram detection at the specific position; the optical fiber left outside the brain is inserted in the ceramic ferrule 4, the optical fiber on the top of the ceramic ferrule 4 is provided with a boundary area which does not cover the insulating film 3, and the boundary area is used for connecting the detection lead 6 with the conductive film through the conductive adhesive 5 so as to realize the derivation of the electroencephalogram signal; the ceramic ferrule 4 is externally sleeved with a ceramic sleeve 7.
The conductive film is made of titanium, platinum, gold and other materials with impedance characteristics and biocompatibility. Preferably, the conductive film is made of a titanium material, has good biocompatibility and corrosion resistance, can stably exist in a living body, and has extremely little rejection reaction; has excellent impedance performance and can conduct the bioelectricity brain signals.
Preferably, the insulating film is made of Parylene (Parylene) materials, has good biocompatibility and insulativity, is suitable for optical fibers implanted in a body for a long time, and can limit the receiving range of electroencephalogram signals.
In the embodiment of the invention, the implanted object of the fiber-optic probe is a mouse with the mass of about 25 to 30g, and the brain volume of the mouse is about 15mm x 8mm x 3mm, wherein 15mm is the longitudinal distance from the forebrain to the hindbrain, 8mm is the transverse distance, and 6mm is the depth. In the research aiming at the Parkinson disease mouse model, the optical fiber needs to carry out light stimulation and electroencephalogram signal acquisition on the mouse substantia nigra. The substantia nigra is located in the mouse brain at a depth of 4mm to 4.1mm, and the size of the substantia nigra is 1.2mm by 1.3mm by 1 mm.
A conductive film is integrally arranged outside the optical fiber; for the part extending into the mouse brain, a detection area with a certain length can be reserved at any position according to research needs and is used for being directly contacted with nerve cells and tissue fluid, and the implanted part except the detection area is coated with an insulating film. The detection zone is covered by a conductive film without an insulating film, and has an axial length of at least 25 microns, preferably 25 to 35 microns, so as to ensure that the detection zone can accurately correspond to a single or a few nerve cells or nerve fibers, and improve the positioning accuracy and the quality of collected signals. When the study object is a mouse suffering from the Parkinson disease, the substantia nigra of the mouse is subjected to light stimulation and electroencephalogram signal acquisition according to the pathology of the Parkinson disease. For the optical fiber probe, the detection area can meet the requirements of treatment and detection on the substantia nigra when the detection area is positioned at the foremost end. Therefore, in the process of this example, the detection area is mainly located at the forefront. It should be noted that the position of the detection area is not limited to the foremost end, and the light stimulation and the acquisition of the fixed-point electroencephalogram signals with different depths can be achieved by changing the arrangement position of the detection area of the part of the optical fiber inserted into the brain. A detection zone having an axial length in the range of 25 to 35 microns may be left at any position of the insertion of the optical fiber into the intracerebral portion, i.e. the detection zone may be located at any position of the implanted portion.
Preferably, the detection wire 6 is a copper wire with the diameter of 0.5mm-1mm, and when the optical fiber is inserted into the ferrule for the condition that the detection zone is positioned at the foremost end (right end) of the optical fiber, a boundary zone which is reserved at the top end of the ferrule and is not covered with an insulating film is used for connecting the copper wire, so that the electroencephalogram signals in a living body can be led out through the copper wire. When the diameter of the copper wire is 0.5mm, the axial length of the interface area is preferably 2mm, and the 2mm area facilitates the connection operation of the conductive adhesive and the copper wire. The following description will be given by way of example of the case where the axial length of the boundary region is 2 mm. The inner diameter of the ceramic ferrule inserted into the optical fiber is matched with the outer diameter of the coated optical fiber, and the ceramic sleeve is matched with the ceramic ferrule, so that the stability of an optical fiber device in the using process is ensured. The diameter of the optical fiber 1 is preferably 200 μm or less, which satisfies the requirement of not damaging the biological tissue.
A method for preparing an optical fiber probe for simultaneously realizing deep brain light stimulation and brain electrical detection comprises the following steps:
s1 plating conductive film on optical fiber
The conductive material is uniformly plated on the optical fiber by utilizing a vacuum coating technology, the conductive film is coated outside the optical fiber, and the impedance of the conductive film meets the following requirements: under the condition of 1kHz, the sum of the impedance of the copper wire and the impedance of the copper wire is less than 500k omega; the electroencephalogram signal can be recorded by using the impedance characteristic of the electroencephalogram signal.
Firstly, cutting an optical fiber with a proper length by using an optical fiber cutting pen or an optical fiber cutting table; observing the section form of the optical fiber respectively in the radial direction and the tangential direction by using a microscope, wherein the section form is as flat as possible so as to ensure the quality of output light spots;
cleaning the optical fiber, sending the optical fiber into a vacuum cavity, introducing high-temperature titanium gas, and adhering titanium to the surface of a workpiece after a certain time to form a uniform titanium film;
and S2, plating an insulating film on the optical fiber.
Uniformly plating Parylene (Parylene) outside the conductive film except the detection area and the boundary area by using a vacuum coating technology; the thicknesses of the conductive film and the insulating film meet the requirement that the diameter of the optical fiber is matched with the inner diameter of the ceramic ferrule after film coating.
For the part extending into the brain of the mouse, the outside of the conducting film is coated by an insulating film except the detection area, the axial length of the detection area is in the range of 25-35 microns, and the validity of the received electroencephalogram signal is ensured; the position of the detection area in the optical fiber intracerebral part can be changed according to the detection requirement. The boundary region for connecting the copper wires is not plated with an insulating film, and when the diameter of the copper wire is 0.5mm, the space is preferably spaced by 2mm in the axial direction. When other diameter copper wires are selected, the distance can be adjusted as desired.
Among them, the insulating film plating method is divided into two types:
the first method is a shielding method: and shielding the two regions which do not need to be plated with the insulating film by using a proper material, and cleaning and pretreating the plated optical fiber. And (3) delivering the treated optical fiber into a vacuum furnace, sublimating, cracking and depositing the raw material on the surface of the test piece to form a layer of compact polymer insulating film.
The second method is a photoetching method: after cleaning the optical fiber, spin-coating photoresist as a mask material on the conducting film for the detection area and the boundary area which do not need to be plated with the insulating film by a chemical vapor deposition method, and then carrying out photoetching to obtain a required sample. The accuracy of this method is usually determined by the resolution of the lithography stage.
The two methods are selected according to actual conditions.
Fig. 1 and 2 show the entire optical fiber and the partial optical fiber in the inspection area after the two steps S1 and S2, in which the inspection area is at the forefront. Fig. 3 shows the overall view of the optical fiber after the first two steps, with the detection zone in the middle.
S3 connection of optical fiber, core insert and sleeve
Clamping the optical fiber to be matched with the ferrule by using an optical fiber clamp, inserting the optical fiber left outside the brain into the ceramic ferrule, and aligning the end part with the ceramic ferrule; sleeving the ceramic sleeve outside the ceramic ferrule and aligning with the ceramic ferrule; therefore, the convenience and the stability of the optical fiber device in the using process are ensured. Fig. 4 shows the positional relationship of the optical fiber, ferrule, and sleeve.
S4 connection of copper wire to conductive film
Clamping the coated optical fiber by using an optical fiber clamp, fixing one end by using an instrument, and connecting a copper wire on the interface area; and in the area with the axial distance of 2mm at the right end of the ceramic ferrule, connecting the copper wire and the conductive film under a microscope. The two copper wires are in central symmetry with the axis of the optical fiber, and the conductive adhesive completely covers the contact point of the copper wires and the conductive film, so that the electroencephalogram signals in a living body can be led out through the copper wires. Fig. 5 shows the relative positions of the wires, conductive glue and optical fibers.
Those skilled in the art will appreciate that the invention may be practiced without these specific details.
Claims (10)
1. An optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection is characterized by comprising an optical fiber (1) for introducing laser under certain conditions, wherein one part of the optical fiber is inserted into the brain of a living body to realize light stimulation; the outside of the optical fiber (1) is coated with a conductive film (2), and the outside of the conductive film (2) is coated with an insulating film (3); a detection area which is not covered by the insulating film (3) is reserved at a specific position on the optical fiber conducting film (2) inserted into the brain of the organism and is used for directly contacting nerve cells and tissue fluid to realize the electroencephalogram detection of the specific position; the optical fiber left outside the brain is inserted in the ceramic ferrule (4), the optical fiber on the top of the ceramic ferrule (4) is provided with a boundary area which is covered with a conductive film but not covered with the insulating film (3), and the boundary area is used for connecting the detection lead (6) with the conductive film through the conductive adhesive (5) so as to realize the derivation of an electroencephalogram signal; and a ceramic sleeve (7) is sleeved outside the ceramic ferrule (4).
2. The optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection according to claim 1, wherein the conductive film (2) is made of a material with impedance characteristics and biocompatibility, and comprises titanium, platinum and gold.
3. The fiber-optic probe for simultaneously realizing deep brain light stimulation and brain electricity detection according to claim 2, characterized in that the preferable material of the conductive film (2) is titanium.
4. The fiber-optic probe for simultaneously realizing deep brain light stimulation and brain electrical detection according to claim 1, wherein the insulating film (3) is Parylene.
5. The fiber-optic probe for simultaneously realizing deep brain light stimulation and brain electrical detection according to claim 1, wherein the axial length of the detection area is at least 25 μm, so as to ensure that a single or a few nerve cells or nerve fibers can be accurately corresponded on the detection area.
6. The optical fiber probe for simultaneously realizing deep brain light stimulation and electroencephalogram detection according to claim 1, wherein the detection lead (6) is a copper wire with the diameter of 0.5mm-1mm, and the axial length of the interface area is at least 2 mm.
7. The fiber probe capable of simultaneously realizing deep brain light stimulation and electroencephalogram detection according to claim 1, wherein the ceramic sleeve (7) is matched with the ceramic ferrule (4), and the inner diameter of the ceramic ferrule is matched with the outer diameter of the coated optical fiber.
8. The optical fiber probe for realizing deep brain light stimulation and brain electricity detection simultaneously as claimed in claim 1, wherein the diameter of the optical fiber (1) is less than 200 microns, so as to meet the requirement of not damaging biological tissues.
9. A method for preparing an optical fiber probe for simultaneously realizing deep brain light stimulation and brain electrical detection is characterized by comprising the following steps:
s1 plating conductive film on optical fiber
Preparing an optical fiber with proper length, uniformly plating a conductive material on the optical fiber by utilizing a vacuum coating technology, coating a conductive film outside the optical fiber, wherein the impedance of the conductive film meets the following requirements: under the condition of 1kHz, the sum of the impedance of the copper wire and the impedance of the copper wire is less than 500k omega; the impedance characteristic of the brain electrical signals can be used for recording the brain electrical signals;
s2 coating the optical fiber with an insulating film
Uniformly plating Parylene outside the conductive film except the detection area and the boundary area by using a vacuum coating technology; the thicknesses of the conductive film and the insulating film meet the requirement that the diameter of the optical fiber is matched with the inner diameter of the ceramic ferrule after film coating;
s3 connection of optical fiber and ceramic ferrule
Inserting the optical fiber left outside the brain into the ceramic ferrule, wherein the end part of the optical fiber is aligned with the ceramic ferrule; sleeving the ceramic sleeve outside the ceramic ferrule and aligning with the ceramic ferrule; therefore, the convenience and the stability of the optical fiber device in the using process are ensured;
s4 connection of copper wire to conductive film
Connecting copper wires on the boundary area; clamping the coated optical fiber by using an optical fiber clamp, and connecting the copper wire and the conductive film under a microscope; the two copper wires are in central symmetry with the axis of the optical fiber, and the conductive adhesive completely covers the contact point of the copper wires and the conductive film, so that the electroencephalogram signals in a living body can be led out through the copper wires.
10. The method for preparing the optical fiber probe for simultaneously realizing deep brain light stimulation and brain electrical detection according to claim 9, wherein in the step S2, the method for plating the insulating film includes two methods:
the first method is a shielding method: shielding a detection area and a boundary area which do not need to be plated with an insulating film by using a proper material, cleaning and pretreating the optical fiber plated with a conductive film, conveying the treated optical fiber into a vacuum furnace, sublimating, cracking and depositing an insulating film raw material on the surface of a test piece to form a layer of compact high-molecular insulating film;
the second method is a photoetching method: after cleaning the optical fiber, spin-coating photoresist as a mask material on the conducting film for the detection area and the boundary area which do not need to be plated with the insulating film by a chemical vapor deposition method, and then carrying out photoetching to obtain a required sample, wherein the precision of the method is usually determined by the resolution of a photoetching link.
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