CN102784010A - Rat primary auditory cortex microelectrode array implantation fixation operation method - Google Patents
Rat primary auditory cortex microelectrode array implantation fixation operation method Download PDFInfo
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Abstract
The invention belongs to the research field of electro-neurophysiology, and particularly relates to a rat primary auditory cortex microelectrode array implantation fixation operation method for auditory sense cognitive research. The method comprises the following steps of: fixing a wire electrode part and an electrode connector part of a microelectrode array in a double ectopia way, i.e. fixing the wire electrode on an auditory cortex temporal bone through an electrode landing device, and then buckling the wire electrode to fix the electrode connector onto a parietal bone so as to complete implantation fixation of the auditory cortex microelectrode array. According to the method, the operation is simple, electrodes can be carried stably, the reliability of signal acquisition is high, the requirement of an auditory sense cognitive research under a freetime state of a rat is met, and the experimental animal cost of correlational researches is reduced.
Description
Technical field
The invention belongs to the neural electrophysiology research field, be specifically related to a kind of rat primary auditory cortex microelectrode array and implant the fixing operation method, to satisfy the needs of under the rat free movement state, gathering its auditory cortex neuron activity.
Background technology
Multichannel the body recording technique can be clear-headed animal, under the free movement state; Record brain regional area colony neuron activity; Be to analyze the strong instrument of colony's neuron, and be widely used in multiple zooperal a plurality of brains district in network level code mechanism.And the multichannel of auditory cortex is carried out on large-scale experiment animals such as Adeps seu carnis Rhiopithecus roxellanae monkey in the research of body record mostly; For the common experimental animal---small-sized rodent such as rat; Only carried out at present under narcotism with passive listen state under the research of (unanesthetized passively listening) colony's neuron activity; Wherein " passive listen state " refers to animal and is in waking state; Listen stimulation but be sitting in the fixture passive reception by training, rather than free active behaving state.And at the research high-level center when listening the cognitive coding of sensory information; Often need of the dynamic change of the relevant brain district of investigation maincenter to the sound IR; On this meaning; The animal behaving state research of the relevant cortex colony of sensation neuron activity down feels that to the detail knowledge maincenter cognitron manufacture-illegal is often important, and does not see the report of Rat Auditory Cortex colony neuron activity under the behaving state at present as yet.Reason is that small test animal primary auditory cortexs such as rat are positioned at brain temporo side; If will implant microelectrode array directly is fixed on the outer temporo laterocranium bone of auditory cortex; Electrode assembly after fixing will cause the variation of animal head normal equilibrium state, and this will influence the free movable of animal.Therefore, under the behaving state Rat Auditory Cortex to implant, firmly carry microelectrode array be to carry out the technical barrier of multichannel in the research of body record.
Generally be divided into wire electrode part and electrode connector (connector is used for realizing and the docking of grapher system) two parts at the active microelectrode array of body collection of record somatic nerves unit.At present; Occurred wire electrode landing gear (landing) on the novel microelectrode array, it and the tight sealing-in of wire electrode can encapsulate dentistry cement earlier around landing when animal head fixed electrode; After treating that the dentistry cement that encapsulates for the first time solidifies fully; Fix at electrode connector encapsulation dentistry cement again, this method available buffer and reduce connector plug grapher system before the vibrations of counter electrode silk when putting (headstage), firm wire electrode.
Implant, firmly carry the technical barrier of microelectrode array to Rat Auditory Cortex under the behaving state; The present invention has carried out design improvement to the microelectrode array that Rat Auditory Cortex is suitable for; Adopt the implantation of carrying out the auditory cortex microelectrode array to the wire electrode of microelectrode array and the dual dystopy fixed method of electrode connector to fix again; Be about to wire electrode and be fixed on auditory cortex temporal bone place, be fixed on electrode connector on the parietal bone behind the bending wire electrode array through electrode landing.This method for implantation relates to the design of the suitable microelectrode array of small test animal auditory cortex, the sport technique segments such as fixing means of auditory cortex microelectrode array.
Summary of the invention
The objective of the invention is to common experimental animal---small-sized rodent such as rat; The microelectrode array that its auditory cortex colony neuron activity of record exists under free movement state is implanted a technique for fixing difficult problem, proposes a kind of rat primary auditory cortex microelectrode array and implants fixed operation method.The rat primary auditory cortex microelectrode array that the present invention proposes is implanted fixed operation method; Be through fixing the wire electrode of microelectrode array part with the dual dystopy of electrode connector part; Be about to wire electrode and be fixed on auditory cortex temporal bone place through the electrode landing gear; Bend wire electrode then and be fixed on electrode connector on the parietal bone, accomplish the implantation of auditory cortex microelectrode array and fix.This method for implantation relates to the design of the suitable microelectrode array of small test animal auditory cortex, the sport technique segments such as implantation fixing means of auditory cortex microelectrode array.
The inventive method is specifically:
Fix through carrying out dual dystopy to wire electrode array and electrode connector; Promptly fix for the first time the wire electrode array is fixed on auditory cortex temporal bone place through electrode landing, fix that for the second time wire electrode array bending back is fixed on electrode connector on the parietal bone.Concrete steps are following:
1) rat brain surgical operation: choose suitable rat; The anesthesia back uses stereotaxic instrument to fix, and before and after the rat head, vertically cuts head scalp, otch 1cm through cerebral surgery operation; Cut off scalp 1cm in otch relief angle vertical left temporo side; Separate temporalis along boss parietal with temporal bone trailing edge passivity, ligation temporo side scalp, temporalis expose parietal bone and left side temporal bone; Use 1% hydrogen peroxide to remove top skull and the outer connective tissue of temporal bone, for follow-up use dentistry cemented electrode is prepared bone surface.
2) screw in fixed screw: corotation is gone into six fixed screws, wherein treats that at temporal bone the both sides, front and back in Kai Nao district respectively pierce a fixed screw, and parietal bone pierces four fixed screws, increases the stable degree of implanting in conjunction with follow-up dentistry cement embedded electrode array.
3) implant microelectrode array at primary auditory cortex: according to the rat brain collection of illustrative plates, use miniature dental burr to open cranium and expose auditory cortex, remove cerebral dura mater, under micro-thruster promotes, insert auditory cortex to microelectrode array.
4) fix the first time after microelectrode array is implanted: microelectrode array is implanted back elder generation dropping agar solution and is covered the auditory cortex surface; Use medical glue (main component is the positive butyl ester of ɑ-alpha-cyanoacrylate) and rarer dentistry cement (to see Fig. 2 more respectively; Dental acrylic layer I) electrode landing and on every side temporal bone closely be fixed together, fix the first time of accomplishing after microelectrode array is implanted.
5) fix the second time after microelectrode array is implanted: after treating that fixed dentistry cement solidifies fully for the first time; Drip normal saline and dissolve the water soluble wax of de-electrifying polar filament array outsourcing; The bending electrod-array makes electrode connector place the animal calvarium, uses more heavy-gravity dentistry cement then; Connector is fixed on the animal calvarium the auditory cortex electrode, and fix the second time of accomplishing after microelectrode array is implanted.
6) postoperative care in the art: note keeping animal heat and anesthesia stable in the art, postoperative carries out disinfection to wound and sews up, and gives intramuscular injection penicillin injection liquid.
The design of the microelectrode array that small test animal auditory cortex wherein is suitable for is following:
1) do not establish electrod driving device, can measure the electrode implantation depth, and can alleviate the weight of electrode assembly, be convenient to the activity of animal head through micro-thruster.
2) increase wire electrode landing gear landing at the electrod-array front end, landing apart from about 10 mm of electrode connector about.
3) select rigidity less, softish nickel filament is as electrode material, outsourcing increases electrod-array hardness with solubility wax.
According to above-mentioned experimental procedure; The adult rat primary auditory cortex is carried out microelectrode array to be implanted and fixes; Actual tests result proves; The dual dystopy fixed method of wire electrode-electrode connector can make microelectrode array for a long time firmly in the Rat Auditory Cortex tracer signal, satisfies the requirement of under the animal free movement state, gathering its auditory cortex colony neuron signal, thereby the auditory cortex microelectrode array of explanation the present invention design and the operation method of use are feasible.The present invention designs and has realized on the active rat of freedom, carrying out the neuronal signal collection of auditory cortex colony; Expand multichannel in the application of body recording technique in auditory sense cognition research, had advantage convenient and easy, that acquired signal reliably reaches reduction correlational study laboratory animal cost.
Description of drawings
Fig. 1 is that small test animal auditory cortex is suitable for the microelectrode array design diagram;
Fig. 2 is that microelectrode array is implanted the fixing means sketch map.
The specific embodiment
Below in conjunction with accompanying drawing the present invention is further described.
As shown in Figure 1, make the microelectrode array that Rat Auditory Cortex is suitable for before the animal surgery earlier, to make the method for microelectrode array according to routine and carry out, particular design wherein is following:
1) do not establish electrod driving device: this design can alleviate the weight of electrode assembly, is convenient to the activity of animal head; In addition,, can measure implantation depth, in implantation process, detect the neuronic reaction of auditory cortex simultaneously and whether touch the hearing reaction neuron with the prompting electrode through electrode micro-thruster implant electrode.
2) increase wire electrode landing gear landing at the electrod-array front end: the landing landing gear on the common in the past microelectrode array is positioned at the wire electrode middle part; The vibrations of counter electrode silk when only playing buffering and reducing electrode connector plug headstage, the effect of firm wire electrode.The auditory cortex microelectrode array landing device that uses among the present invention is positioned at the about 3 mm places of eletrode tip, landing apart from about 10 mm of electrode connector about, satisfy the wire electrode array is bent to parietal bone from auditory cortex length requirement.Special according to Rat Auditory Cortex in the body position; The Position Design of this landing also is to implant fixed indispensable structure through carrying out wire electrode-connector dystopy double fixed realization microelectrode array at auditory cortex in the operation method of the present invention except playing the purpose of firm wire electrode.
3) select rigidity less, softish nickel filament is as electrode material, be easy to the subsequent procedures operation from auditory cortex bending wire electrode array to parietal bone; Outsourcing increases electrod-array hardness with solubility wax, passes pia mater encephali to make things convenient for electrod-array.
This microelectrode array is divided into electrode interface 1 (microelectrode array connector) and wire electrode array 2 (microelectrode array) two parts among Fig. 1; Wherein locate to be provided with electrode landing gear 3 (microelectrode array landing) at most advanced and sophisticated 3 mm of distance electrode silk (L2); Wire electrode landing is apart from following 10 mm of electrode interface (L1), wire electrode array total length (L1+L2) 13 mm.
The implantation of Rat Auditory Cortex microelectrode array is fixed:
Select adult SD rats, male and female are not limit, available from the Zhejiang Academy of Medical Sciences animal center.After animal is bought back, sub-cage rearing at random.The comfortable environment of keeping quite between feeding period lets it freely drink water and take food.Carry out audition before the operation and detect, the normal person of hearing reaction carries out the implant surgery of auditory cortex microelectrode array.The microelectrode array that uses Rat Auditory Cortex to be suitable for, by following step successively anaesthetize, cerebral surgery operation exposes primary auditory cortex, implant the laggard column electrode silk of microelectrode array-dual dystopy of electrode connector fixes, its concrete steps process is:
1) rat brain surgical operation: after rat is weighed; Through lumbar injection pentobarbital sodium (sodium pentobarbital; 40 mg/kg body weight) anesthesia gives subcutaneous injection atropine (atropine sulfate, 0.07mg/kg body weight) afterwards and prevents that trachea, bronchus from producing secretions.Use fixedly rat of 45 ° of ear bar stereotaxic instruments, cut off the rat head by hair and left temporo side by hair, the head scalp is vertically cut in front and back, otch 1cm, exposed tops skull; Otch relief angle vertical left temporo side is cut off scalp 1cm at the top, separates temporalis along boss parietal with temporal bone trailing edge passivity, ligation temporo side scalp, temporalis, and tractive ligature expansion temporo side operating field exposes auditory cortex outside temporal bone; Use 1% hydrogen peroxide to remove top skull and the outer connective tissue of temporal bone, for follow-up use dentistry cemented electrode is prepared bone surface.
2) screw in fixed screw: at first according to the rat brain collection of illustrative plates, solid is located primary auditory cortex, and marks the position of auditory cortex with pencil.Use miniature dental burr and diameter 0.5 mm cylindrical drill to bore an aperture respectively in the both sides, front and back of treating the Kai Nao district respectively, be used to screw in fixed screw; Uniform distribution bores five apertures around the parietal bone that exposes again, and wherein four apertures are used for fixed screw on every side, screw in six fixed screws altogether, in conjunction with follow-up dentistry cement embedding fixed screw, can increase the stable degree that electrode is implanted.In addition, treat that near auditory cortex an aperture in Kai Nao district is used to insert reference electrode on the parietal bone.
3) implant microelectrode array at primary auditory cortex: according to three-dimensional telltale mark point; Temporal bone bores out an aperture (about 2.5 mm of diameter) gently with miniature dental burr in the auditory cortex outside; Remove cerebral dura mater; After cleaning out the brain surface with normal saline; 8 electrodes that promote following ready 8 road electrod-arrays at micro-thruster carefully insert auditory cortex and in the reaction of body record neuron, judge according to the implantation depth and the neuronic discharge characteristics of auditory cortex of micro-thruster whether eletrode tip arrives auditory cortex, stop during to auditory cortex IV layer advancing.
4) fix the first time after microelectrode array is implanted: get agar solution (concentration 2%) with 20 μ l liquid-transfering guns and cover the auditory cortex surface; Treat that agar solidifies the back and forms the layer of gel film; Physiological status with the protection cortex; Use medical glue (main component is the positive butyl ester of ɑ-alpha-cyanoacrylate) to be coated on electrode landing and the temporal bone then, treat to modulate the rarer dentistry cement of concentration (dental acrylic layer I) again after the medical glue drying, get dentistry cement drop at wire electrode landing and surrounding cranial bone with 20 μ l liquid-transfering guns; Closely be fixed together wire electrode landing and surrounding cranial bone, fix the first time of accomplishing after microelectrode array is implanted.
5) fix the second time after microelectrode array is implanted: after treating that fixed dentistry cement solidifies fully for the first time, drip the molten solubility wax that goes of normal saline.The bending electrod-array; Make electrode connector place the animal calvarium; Modulate more heavy-gravity dentistry cement (dental acrylic layer II) then, connector is fixed on the animal calvarium the auditory cortex electrode, and fix the second time of accomplishing after microelectrode array is implanted.
6) postoperative care in the art: at last wound is carried out disinfection and sew up, give intramuscular injection penicillin injection liquid.Note keeping animal heat and anesthesia stable in the art.
7) recording electrode load time; And use Plexon multi-channel nerve signal analysis and processing system (Multichannel Acquisition Processor System, MAP System) to carry out in the body signals collecting in different time points (online acquisition during implant surgery, postoperative 3 days, 1 week of postoperative, two 2 weeks of postoperative, postoperative January, postoperative February).The stability of inspection operation method of the present invention and the reliability of signals collecting.
8) put to death animal and carry out the formalin perfusion postoperative bimester, the brain frozen section shows the whether normal in the body position of microelectrode array.
Among Fig. 2; Microelectrode array 11 the rat primary auditory cortex implant after electrode landing gear 9 after auditory cortex temporal bone place uses dentistry cement 7 to carry out fixing first time; Bending wire electrode array uses dentistry cement 6 to carry out the second time through electrode interface 5 and fixes to parietal bone.Wherein, 10 agar gel films for sealing auditory cortex surface, 12 is the rat skull.Can insert multi-channel nerve signal analysis and processing system 8 (Multichannel Acquisition Processor System, MAP System) through holding wire 4 after electrode is implanted and fixed carries out in the body signals collecting.
Claims (2)
1. rat primary auditory cortex microelectrode array is implanted the fixing operation method, it is characterized in that this method may further comprise the steps:
Fix through carrying out dual dystopy to wire electrode array and electrode connector; Promptly fix for the first time the wire electrode array is fixed on auditory cortex temporal bone place through electrode landing; Fixing for the second time with being fixed on electrode connector on the parietal bone after the bending of wire electrode array, concrete steps are following:
Step (1) rat brain surgical operation: choose suitable rat; The anesthesia back uses stereotaxic instrument to fix, and before and after the rat head, vertically cuts head scalp, otch 1cm through cerebral surgery operation; Cut off scalp 1cm in otch relief angle vertical left temporo side; Separate temporalis along boss parietal with temporal bone trailing edge passivity, ligation temporo side scalp, temporalis expose parietal bone and left side temporal bone; Use 1% hydrogen peroxide to remove top skull and the outer connective tissue of temporal bone, for follow-up use dentistry cemented electrode is prepared bone surface;
Step (2) screws in fixed screw: corotation is gone into six fixed screws, wherein treats that at temporal bone the both sides, front and back in Kai Nao district respectively pierce a fixed screw, and parietal bone pierces four fixed screws, increases the stable degree of implanting in conjunction with follow-up dentistry cement embedded electrode array;
Step (3) is implanted microelectrode array at primary auditory cortex: according to the rat brain collection of illustrative plates, use miniature dental burr to open cranium and expose auditory cortex, remove cerebral dura mater, under micro-thruster promotes, insert auditory cortex to microelectrode array;
Fix the first time after step (4) microelectrode array is implanted: microelectrode array is implanted back elder generation dropping agar solution and is covered the auditory cortex surface; Use respectively more medical glue and rarer dentistry cement electrode landing and on every side temporal bone closely be fixed together, fix the first time of accomplishing after microelectrode array is implanted;
Fix the second time after step (5) microelectrode array is implanted: after treating that fixed dentistry cement solidifies fully for the first time; Drip normal saline and dissolve the water soluble wax of de-electrifying polar filament array outsourcing; The bending electrod-array makes electrode connector place the animal calvarium, uses more heavy-gravity dentistry cement then; Connector is fixed on the animal calvarium the auditory cortex electrode, and fix the second time of accomplishing after microelectrode array is implanted;
Postoperative care in step (6) art: note keeping animal heat and anesthesia stable in the art, postoperative carries out disinfection to wound and sews up, and gives intramuscular injection penicillin injection liquid.
2. rat primary auditory cortex microelectrode array according to claim 1 is implanted the fixing operation method, it is characterized in that:
The concrete design of microelectrode array is following:
Steps A. do not establish electrod driving device, can measure the electrode implantation depth, and can alleviate the weight of electrode assembly, be convenient to the activity of animal head through micro-thruster;
Step B. increases wire electrode landing gear landing at the electrod-array front end, landing apart from about 10 mm of electrode connector about;
Step C. select rigidity less, softish nickel filament is as electrode material, outsourcing increases electrod-array hardness with solubility wax.
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| CN103690262A (en) * | 2013-11-29 | 2014-04-02 | 南京航空航天大学 | Electrode adapter for avian chronic brain function researches and fixing method |
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| CN103445790A (en) * | 2013-09-09 | 2013-12-18 | 重庆西南医院 | Animal auditory detecting electrode positioning device |
| CN103690262A (en) * | 2013-11-29 | 2014-04-02 | 南京航空航天大学 | Electrode adapter for avian chronic brain function researches and fixing method |
| CN103690262B (en) * | 2013-11-29 | 2016-01-20 | 南京航空航天大学 | Birds chronic cerebral functional study electrode adapter |
| CN105852855A (en) * | 2016-04-15 | 2016-08-17 | 郑州科斗创客科技有限公司 | Implantable cerebral electrode for measuring cerebral primary visual electrocorticograms in rodents |
| CN108742942A (en) * | 2018-06-19 | 2018-11-06 | 中国科学院电子学研究所 | General anesthesia neuromechanism studies acute experiment platform and chronic experiment platform |
| CN110786846A (en) * | 2019-12-11 | 2020-02-14 | 中国科学院深圳先进技术研究院 | Flexible recording electrode and preparation method and implantation method thereof |
| CN110786846B (en) * | 2019-12-11 | 2024-02-20 | 中国科学院深圳先进技术研究院 | Flexible recording electrode and preparation method and implantation method thereof |
| CN114305335A (en) * | 2021-12-24 | 2022-04-12 | 中国科学院空天信息创新研究院 | Wear-type many brain areas neural detection regulation and control microsystem and receive electrode array module a little |
| CN114305335B (en) * | 2021-12-24 | 2023-10-13 | 中国科学院空天信息创新研究院 | Head-wearing type multi-brain-area nerve detection regulation and control micro-system and micro-nano electrode array module |
| CN114847224A (en) * | 2022-04-18 | 2022-08-05 | 华东师范大学 | Device and method for detecting sound frequency resolution capability of mouse |
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Application publication date: 20121121 |