CN103948452A - Inhalation-anesthesia exposing and monitoring device - Google Patents
Inhalation-anesthesia exposing and monitoring device Download PDFInfo
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Abstract
The invention provides an inhalation-anesthesia exposing and monitoring device, which comprises an oxygen or air-oxygen mixed-gas tank, an inhalation-anesthetic volatilizing tank, carrier-gas pipes, a gas inlet hole, an inhalation-anesthetic exposing box, a gas outlet hole, an anesthetic-gas adsorber used in a loop, a multi-parameter anesthetic-gas monitor, an animal body-temperature maintainer, a line, a line access hole, a dissecting microscope, a constant-temperature heating plate, a small-mouse fixing hole, an invasive arterial blood-pressure monitor, a heat-preserving blanket and an operating table, wherein the oxygen or air-oxygen mixed-gas tank, the inhalation-anesthetic volatilizing tank, the inhalation-anesthetic exposing box and the anesthetic-gas adsorber are connected with the multi-parameter anesthetic-gas monitor through the carrier-gas pipes, and the animal body-temperature maintainer is connected with the heat-preserving blanket. The experiments of a plurality of large/small mice can be simultaneously carried out with sufficient and sensitive heat preservation, real-time monitoring and self-heating, and the breathing conditions of the large/small mice under an anesthetic state are observed at any moment. The inhalation-anesthesia exposing and monitoring device has the advantages of reasonable design, simplicity and convenience in operation and strong controllability and not only has the function of anesthetic-concentration monitoring, but also is provided with an anesthetic absorbing device, so that the inhalation-anesthesia exposing and monitoring device is safe and environmentally friendly.
Description
Technical field
The invention belongs to animal experiment device capable, relate to a kind of inhalation anesthesia and expose and monitoring device, be applicable to temperature monitoring, the arterial blood gas that rat/mouse carries out in inhalation anesthesia process-exposed and have the monitoring of wound arteriotony.
Background technology
In research Inhalation Anesthetic long-time (> 0.5h) exposes the experiment of rat/mouse impact, be bound to relate to the problem of exposing device, in the middle of common inhalation anesthetic exposes experiment, the in the situation that of conditions permit, can keep the autonomous respiration of animal and avoid animal to carry out tracheal intubation or mechanical ventilation is carried out in tracheotomy.1. the main following points of reason of selecting to keep animal autonomous respiration and do not carry out tracheal intubation carry out tracheal intubation or tracheotomy operation, increase experimental implementation step, extend experimental period and 2. carry out tracheal intubation or tracheotomy, because experimental implementation is lost blood, airway secretions, conduit distortion, the reason such as catheter position is inaccurate can cause the mortality rate of laboratory animal to increase.3. carry out tracheal intubation or tracheotomy, increase the air flue loss to animal, cause the stress that operation stimulates, and the factor such as lose blood of operation process, increase the uncontrollable factor of experimentation, easily affect experimental result.But select to keep the mode of animal autonomous respiration just to need special anesthesia exposing device, this device must satisfied condition be to keep under the state of animal autonomous respiration, laboratory animal is carried out to body temperature control, rat/mouse body temperature under requirement of experiment narcotism must remain on 37 ± 0.5 DEG C, because hyperpyrexia or too low every physiological status that all can have influence on mice is stable, affect experimental result, and can increase the mortality rate of laboratory animal.And, in some experiment (if inhalation anesthetic is on the neurovirulent impact experiment of rat/mouse), in order to get rid of hypoxia, high carbon dioxide and hypotension, the factors such as hypertension need to be carried out arterial blood gas analysis and have wound arteriotony to monitor to maintain soda acid the rat/mouse in anesthesia process-exposed the impact of experimental result, electrolyte, stablizing of oxygenate and circulation, this just has higher requirement to anesthesia exposing device, require this device to expose and maintenance body temperature in the situation that ensureing that rat/mouse normally anaesthetize, carry out anesthetic concentration monitoring, the sampling monitoring arterial blood gas of arterial blood, and there is wound arteriotony to monitor.Our laboratorys are published in the document Physiological disturbance may contribute to neurodegeneration induced by isoflurane or sevoflurane in 14 day old rats. on PLoS One magazine and explicitly point out in January, 2014, and Physiological factors abnormal as low temperature, hypoxia, high carbon dioxide, abnormal blood pressure etc. can be to laboratory animal generation neurodevelopment toxicity.
Common used inhalation anesthetic exposing device is as Fig. 3 at present, its Insulation is anesthesia to be exposed to case be positioned in 37 DEG C of waters bath with thermostatic control, certain like this insulation problem that solved, but the seal that exposes case for anesthesia has proposed requirements at the higher level, must can not leak gas and leak completely, especially must just can ensure through special handling at the seam of carrier gas channel discrepancy chest.Topmost problem is, in this case, can not in the middle of anesthesia process-exposed, carry out the extraction of arterial blood and have the monitoring of wound arteriotony animal, animal must be taken and expose outside case or exposure case taken out to water-bath and just can carry out, and the guarantee of such temperature that just can not ensure inhalation anesthesia concentration or lose, measured next vim and vigour or arteriotony can not really reflect vim and vigour and the blood pressure in Animal Anesthesia process-exposed in this case.For these problems, we have invented this experimental provision system.
Summary of the invention
The object of the invention is to overcome the conventional deficiency that sucks exposing device at present, solve the unsolved problem of device at present, provide a kind of inhalation anesthesia to expose and monitoring device, this device is by oxygen or empty oxygen gas mixture tank, inhalation anesthetic volatilization tank, carrier gas pipe, air inlet, inhalation anesthetic exposes case, venthole, in loop, use anesthetic gas absorber, multiparameter anaesthetic gas monitor, animal heat maintains instrument, circuit, circuit access hole, anatomic microscope, constant temperature heating plate, mice fixing hole, there is wound arteriotony monitor, insulation blanket, operating board composition, air inlet is opened in the middle of inhalation anesthetic exposes case right side, venthole is opened above inhalation anesthetic exposes case left side, circuit access hole is opened below inhalation anesthetic exposes case left side, mice fixing hole is opened below inhalation anesthetic exposes case front, airtight inhalation anesthetic exposes case and is placed on operating board, oxygen or empty oxygen gas mixture tank connect inhalation anesthetic volatilization tank through carrier gas pipe, connect inhalation anesthetic through air inlet again and expose case, in loop, use anesthetic gas absorber to be connected inhalation anesthetic with multiparameter anaesthetic gas monitor through venthole and expose case, animal heat maintains instrument and is connected with insulation blanket through access hole by power line, insulation blanket is placed in anesthetics and exposes case bottom, maintain instrument control insulation blanket temperature processed by animal heat, constant temperature heating plate is positioned at mice fixing hole below, anatomic microscope is placed on operating board, near constant temperature heating plate and mice fixing hole, there is wound arteriotony monitor to be placed on operating board, in the time that having the monitoring of wound arteriotony, needs use is connected with the tremulous pulse of laboratory animal (rat/mouse) by tremulous pulse pressure-measuring pipe.
It is completely airtight that inhalation anesthetic exposes case, material is lucite material, the better tightness at edge, owing to not needing to be placed in water bath, there is not seepy question, chest is transparent, breathing state in activity and the experimental animal rat/mouse anaesthesia process of animal in inspection box easily, be shaped as cuboid, its size is according to the size design of insulation blanket, just insulation blanket can be placed in and expose case bottom, below the left side of chest, have circuit access hole, can pass through by power supply source line, the gap of power circuit and lucite is by adhesive closure, guarantee can not leak gas.Power line in insulation blanket maintains instrument with animal heat and is connected, and there is probe for detecting temperature can put into rat/mouse rectum monitoring rat/mouse body temperature, the temperature of insulation blanket can regulate so that rat/mouse body temperature reaches 37 degrees Celsius of the temperature that set according to rat/mouse body temperature, can control accurately like this rat/mouse body temperature.This is one of innovative point of the present invention.In the time that exposing, animal inhalation anesthetic animal bodies can be placed in insulation blanket, only the head of animal is stretched out outside insulation blanket, so not only can ensure animal heat but also can complete inhalation anesthesia to expose, can observe by the undulating state of animal chest the breathing state of animal simultaneously.In addition, the sodica calx that tiles on insulation blanket, can better absorb carbon dioxide and the water vapour that animal is breathed out like this.The junction that it is carrier gas channel that air inlet is opened in the centre position, right side of inhalation anesthetic exposure case, oxygen or empty oxygen gas mixture and anesthetic gases thus hole enter in case, and the position in hole facilitates the even diffusion of gas in centre.Above the left side of chest, be venthole, outlet pipe is externally connected with in exhaust gas absorption device loop and uses anesthetic gas absorber, avoids waste gas to enter in air and pollutes; There is multiparameter anaesthetic gas monitor in the nearly case termination of escape pipe, can adjust accordingly the flow that enters anesthesia gas concentration and gas carrier, guarantee to anaesthetize the stable of the interior anesthesia gas concentration of process-exposed raising middle flask simultaneously.
This device inhalation anesthetic expose case above below have the rat/mouse fixing hole that diameter is 2cm, this hole is closed in the time that animal is normally anaesthetized exposure, and at needs, experimental animal rat/mice is carried out to arterial blood gas analysis and have wound arteriotony when monitoring open, this hole just allows the head of animal to enter, and health is outside chest, can in the time that exposing, anesthesia carry out like this real-time blood gas analysis and arteriotony monitoring.In the time of the arterial blood gas real-time analysis of anaesthetizing while exposing, select experimental animal rat/mice to carry out arterypuncture blood drawing instead of cardiac puncture carries out arterial blood gas sampling, be because mouse heart very little if to carry out the blind difficulty of wearing larger, the position of puncture is difficult for determining, success rate is low, and constant error penetrates right ventricle and extracts venous blood out.Easily cause pneumothorax to affect Results of Blood-gas if open the lower puncture of breast direct-view left ventricle, therefore selecting extraction arterial blood more convenient operation is simple, after use apparatus of the present invention, more easily completes.Simultaneously in order also to ensure that the body temperature of animal is constant in the time doing these operations, select animal to be placed in the enterprising line operate of constant temperature heating plate, if some action need carries out under anatomic microscope, also can hot plate be placed under microscope simultaneously, operate very simple and convenient.
This device simplicity of design, easy to operate, can allow many rat/mouse to carry out anaesthetic exposure simultaneously, be incubated fully sensitively, digitized shows and body temperature is carried out to Real-Time Monitoring independently heats there is no particular/special requirement to exposing case.Casing is originally as pmma material, and appearance is transparent, facilitates experimenter to observe the breathing situation under animal activity and narcotism.The aperture design of chest below has met the requirement that animal is operated under narcotism, and hot plate when operation has ensured animal heat, simple to operate, can allow to operate without any the experimenter of experience.Whole device is reasonable in design, simple to operation, and controllability is strong, ensures that experimentation is smooth and easy, and existing anesthetic concentration monitoring, has again anaesthetic absorption plant, safety and environmental protection, practicality and highly versatile.
brief description of the drawings
Fig. 1 is the structural representation of inhalation anesthesia exposure of the present invention and monitoring device.
Fig. 2 is the structural representation of inhalation anesthesia exposure of the present invention and monitoring device embodiment.
Fig. 3 is the structure chart of traditional inhalation anesthetic exposing device.
Detailed description of the invention
The present invention in conjunction with the accompanying drawings and embodiments, is described further.
embodiment 1a kind of inhalation anesthesia exposes and monitoring device
Referring to Fig. 1, Fig. 2, this device is by oxygen or empty oxygen gas mixture tank 1, inhalation anesthetic volatilization tank 2, carrier gas pipe 3, air inlet 4, inhalation anesthetic exposes case 5, venthole 6, in loop, use anesthetic gas absorber 7, multiparameter anaesthetic gas monitor 8, animal heat maintains instrument 9, connect the circuit 10 that insulation blanket and animal heat maintain instrument, circuit access hole 11, anatomic microscope 12, constant temperature heating plate 13, mice fixing hole 14, there is wound arteriotony monitor 15, insulation blanket 17, operating board 19 forms, air inlet 4 is opened in the middle of inhalation anesthetic exposes case 5 right sides, venthole 6 is opened above inhalation anesthetic exposes case 5 left sides, circuit access hole 11 is opened below inhalation anesthetic exposes case 5 left sides, mice fixing hole 14 is opened below inhalation anesthetic exposes case 5 fronts, airtight inhalation anesthetic exposes case 5 and is placed on operating board 19, oxygen or empty oxygen gas mixture tank 1 connect inhalation anesthetic volatilization tank 2 through carrier gas pipe 3, connect into anesthetics through air inlet 4 again and expose case 5, in loop, use anesthetic gas absorber 7 to be connected inhalation anesthetic with multiparameter anaesthetic gas monitor 8 through venthole 6 and expose case 5, waste gas is entered in loop and is used anesthetic gas absorber 7 to be absorbed by venthole 6, multiparameter anaesthetic gas monitor 8 is monitored inhalation anesthetic substrate concentration in waste gas, animal heat maintains instrument 9 and is connected with insulation blanket 17 through access hole 11 by circuit 10, insulation blanket 17 is placed in inhalation anesthetic and exposes case 5 bottoms, maintain instrument 9 by animal heat and control insulation blanket 17 temperature, constant temperature heating plate 13 is positioned at mice fixing hole 14 belows, anatomic microscope 12 is placed on operating board 19, near constant temperature heating plate 13 and mice fixing hole 14, because mice tremulous pulse more carefully can operate for 12 times at anatomic microscope, there is wound arteriotony monitor 15 to be placed on operating board 19, in the time that having the monitoring of wound arteriotony, needs use is connected with the tremulous pulse of laboratory animal (rat/mouse) by tremulous pulse pressure-measuring pipe.
Rat/mouse 16 inhalation anesthesias are placed in the interior maintenance thermostasis of insulation blanket 17 of inhalation anesthetic exposure case 5 while exposure, insulation blanket is covered with sodica calx 18 outward for absorbing the CO of waste gas
2if carry out rat/mouse arterial blood gas and have the monitoring of wound arteriotony, rat/mouse being taken out rapidly to inhalation anesthetic exposes case 5 and the head of rat/mouse is put into inhalation anesthetic exposure case 5 through mice fixing hole 14, mice health is placed on simultaneously and on constant temperature heating plate 13, carries out operation technique, because mice tremulous pulse more carefully can operate for 12 times at anatomic microscope, after arterypuncture success, by there being the tremulous pulse pressure-measuring pipe in wound arteriotony monitor 15 to insert mice tremulous pulse, carry out arteriotony monitoring.This device is easy and simple to handle, is easy to grasp and operation.
It is completely airtight that this inhalation anesthetic exposes case 5, and making material is lucite material, and the better tightness at edge, owing to not needing to be placed in water bath, does not exist seepy question.Chest is transparent, easily the breathing state in activity and the anaesthesia process of animal in inspection box.It is cuboid that inhalation anesthetic exposes case 5, its size is according to the size design of insulation blanket 17, just insulation blanket 17 can be placed in and expose case bottom, below the left side of chest, have power circuit access hole 11, can hold power circuit 10 passes through,, by adhesive closure, guarantee can not leak gas in the gap of power circuit 10 and lucite.Animal heat maintains instrument 9 and is connected with insulation blanket 17 through access hole 11 by circuit 10, and there is temperature probe can put into rat/mouse rectum monitoring mouse temperature, the temperature of insulation blanket can regulate so that rat/mouse body temperature reaches 37 degrees Celsius of the temperature that set according to rat/mouse body temperature, can control accurately like this rat/mouse body temperature.
This device has the rat/mouse fixing hole 14 that diameter is 2cm below the front of inhalation anesthetic exposure case 5, this hole is closed in the time that animal is normally anaesthetized exposure, and at needs, experimental animal rat/mice is carried out to arterial blood gas analysis and have wound arteriotony when monitoring open, this hole just allows the head of animal to enter, and health is outside chest, can in the time that exposing, anesthesia carry out like this real-time blood gas analysis and arteriotony monitoring.In the time of the arterial blood gas real-time analysis of anaesthetizing while exposing, select experimental animal rat/mice to carry out arterypuncture blood drawing instead of cardiac puncture carries out arterial blood gas sampling, be because mouse heart very little if to carry out the blind difficulty of wearing larger, the position of puncture is difficult for determining, success rate is low, and constant error penetrates right ventricle and extracts venous blood out.Easily cause pneumothorax to affect Results of Blood-gas if open the lower puncture of breast direct-view left ventricle, therefore selecting extraction arterial blood more convenient operation is simple, after use apparatus of the present invention, more easily completes.Simultaneously in order also to ensure that the body temperature of animal is constant in the time doing these operations, select animal to be placed in the enterprising line operate of constant temperature heating plate 13, if some action need carries out for 12 times at anatomic microscope, also can hot plate 13 be placed under microscope simultaneously, operate very simple and convenient.
In the time that Animal Anesthesia exposes, animal bodies can be placed in insulation blanket, only the head of animal be stretched out outside insulation blanket, so not only can ensure animal heat but also can complete anesthesia to expose, can observe by the undulating state of animal chest the breathing state of animal simultaneously.In addition, there is sodica calx 18 to be laid in dish in the both sides of insulation blanket 17, can better absorb like this carbon dioxide and the water vapour that animal is breathed out.In the middle of the right side of inhalation anesthetic exposure case 5, have air inlet 4, have oxygen or empty oxygen gas mixture and anesthetic gases to enter in case in hole thus by carrier gas pipe 3, the position of air inlet 4 facilitates the even diffusion of gas in centre.Above the left side of chest, be venthole 6, connect anesthetic gas absorber 7, form exhaust gas absorption device loop, avoid waste gas to enter in air and pollute; There is multiparameter anaesthetic gas monitor 8 in the nearly case termination of venthole 6, can adjust accordingly the flow that enters anesthesia gas concentration and gas carrier, guarantee to anaesthetize the stable of the interior anesthesia gas concentration of process-exposed raising middle flask simultaneously.
Compared with traditional rat/mouse inhalation anesthesia exposing device, the present invention has changed over traditional water bath heat preserving method the insulation blanket of temperature automatically controlled insulation, be connected by maintaining instrument with animal heat, can be better by the monitoring of anus temperature maintain the thermostasis of animal in inhalation anesthesia process, easy to operate.The exposing below chest and offer aperture at inhalation anesthesia of novelty of the present invention in addition, for passing through the head or tail portion of rat/mouse, convenient operations in animal inhalation anesthesia process-exposed, has solved the problem that animal is difficult to carry out various experiment processing under inhalation anesthesia exposes.Simultaneously, we provide and rat/mouse have been carried out under inhalation anesthesia to body temperature, arterial blood gas, there is a set of complete operating means system of wound arteriotony monitoring, except prior device, we have also increased zootomy microscope, animal constant temperature hot plate, there are the devices such as wound arteriotony monitor, these devices have solved some difficult problem problems that may run in experimentation, as mice tremulous pulse too carefully carries out the high problem of arterypuncture success rate low technical, the insulation problem of animal in the time accepting experimental implementation processing etc., reduce experimental implementation difficulty, can make abecedarian can both complete experimental implementation.
embodiment 2application apparatus of the present invention are carried out adult mice autonomous respiration and are kept homoiothermal anesthesia to expose experiment
Before experiment, prepare: with reference to Fig. 2, that the autonomous respiration of a kind of maintenance adult mice keeps homoiothermal inhalation anesthesia exposing device, before Animal Anesthesia medicine exposure prerequisite, animal heat being maintained to instrument 9 opens, it is 37 DEG C that steady temperature is set, because experiment mice needs to keep thermostasis at 37 ± 0.5 DEG C under narcotism.Experiment starts front testing fixture seal, and determining device seal is good.Tremulous pulse body temperature maintains instrument 9 and is purchased from the model SS20-2 of Huaibei Zhenghua Biological Instrument Co., Ltd. of Anhui Province.
Induction of anesthesia: 3 C57BL/6J mice 16 body weight are born the 2-3 month between 25g-33g, put into anesthesia and expose case 5 beginning induction of anesthesia, open oxygen tank 1, adjust flux is 2L/min, open inhalation anesthetic isoflurane volatilization tank 2, regulating concentration is 2.5%, opens multiparameter anaesthetic gas monitor 8 simultaneously Exhaust Gas anesthetic concentration is carried out to Real-Time Monitoring, until experiment mice, anyway when areflexia, induction of anesthesia completes.Multiparameter anaesthetic gas monitor 8 is Drager model Vamous.
Anesthesia maintains: rapidly mice 16 healths are put into insulation blanket 17 and the probe for detecting temperature that animal heat maintains instrument 9 is placed in to mice internal rectum fixing, keep mice 16 heads to stay outside insulation blanket, and sodica calx 18 is laid on the surface of insulation blanket, contribute to absorb carbon dioxide and the steam of discharging in gas.It is 1.4% that inhalation anesthetic isoflurane concentration is turned down, and the moment observe discharge aspiration enter anesthetic concentration, it is also maintained to 1.4%, the waste gas of discharge, by using anesthetic gas absorber 7 to absorb in loop, prevents that waste gas from entering air and polluting.In anesthesia maintenance process, close observation mouse breathing, and be recorded in respectively the mouse breathing frequency of anaesthetizing while maintaining 10min, 30min, 1h, 1.5h, 2h, tentatively judge whether mice respiration inhibition has occurred.Anaesthetize the common 2h that holds time.In loop, use anesthetic gas absorber 7(Anesthetics Gas Absorber, AGA) be the model XF-2 of Nanjing Tian Ao Medical Instruments Manufacturing Co., Ltd.
There are wound tremulous pulse pressure measurement and arterial blood gas analysis: maintaining 1h, 1.5h, 2h in anesthesia respectively has wound tremulous pulse pressure measurement and extract arterial blood and carry out arterial blood gas analysis a mice wherein.Detailed process is as follows: get rapidly a mice outside case, and mouse head is put into mouse head fixing hole 14, and mice health is fixed on constant temperature heating plate 13 with adhesive tape, so just can ensure that mice carries out normal anesthesia and exposes and keep extracting mice arterial blood in homoiothermal situation simultaneously and carry out arterial blood gas analysis, and can monitor mice simultaneously and have wound arteriotony.Due to mice small volume, mice ventral aorta diameter only has 0.6-0.9mm thick, and under naked eyes, to carry out ventral aorta puncture difficulty larger for diameter, and therefore we are chosen in anatomic microscope and operate for 12 times, the anatomic microscope that we use is OLYMPUS model SZ61,10 times of amplifications.After anesthetic concentration is stable, because the inhalation anesthetic isoflurane concentration 1.4% of selecting is mice 1MAC value, because will dissect operation to mice, stimulate larger, can affect other mouse anesthesia states in case and increase isoflurane concentration, therefore stimulate volume 0.3ml after diluting to mouse peritoneal injection ketamine 2.5mg/kg before operation in order to reduce operation technique.After waiting for 5min ketamine (ketamine is analgesic effect) onset herein, start to dissect operation, operating process small mouse is moving without body.Under microscope, cut mice left lower abdomen skin, subcutaneous tissue, push the tissue such as intestinal tube, stomach fat aside, find ventral aorta along spinal column, ventral aorta is closely connected with caval vein, the thick peony of caval vein is easy to discovery, and ventral aorta, just in venacaval left side, is beaten obviously.The two surperficial tunicle is too many, not easily separated, easily punctures vein when separation, therefore selects only ventral aorta surface tunicle to be divided totally, approximately can separate 1cm length, surveys each folder one bulldog clamp in tremulous pulse head side and tail.Puncture with 24g arterial puncture needle at mid portion, after the successful backed off after random nook closing member that punctures connects tremulous pulse pressure-measuring pipe, open head end bulldog clamp, carry out tremulous pulse pressure measurement, continue, after pressure measurement 10min, to take out arterial blood and carry out blood gas analysis.Having wound tremulous pulse monitor 15 is PHILIPS model M3046A.Blood gas analyzer is Gem premier 3000.
According to above method, do altogether 18 mices, relevant parameter is referring to table 1, table 2.
By data in two tables, we may safely draw the conclusion, use device of the present invention, can complete and keep autonomous respiration to keep homoiothermal inhalational anesthetic exposure to adult mice, and can be very easily to the arterial blood gas in mouse anesthesia process-exposed and there is wound arteriotony to monitor, and the breathing situation that can simultaneously observe mice, anaesthetize process-exposed in this device in, there is not respiration inhibition in digital proof mice, change of respiratory rate is little, PH, the PaCO of mice
2,arteriotony is all within normal range, and mice, in anesthesia process-exposed, the abnormal physiology situations such as acidosis, alkalosis, hypotension, hypertension does not occur.
embodiment 3a kind of apparatus of the present invention of applying keep adult mice autonomous respiration to keep the experiment of under the exposure of thermostasis inhalation anesthesia, inhalation anesthetic MAC being measured
Before experiment, prepare: with reference to Fig. 2, this experiment difference is that constant temperature heating plate is put into anesthesia exposes case 5, and power line is stretched out by circuit access hole 11, connects power supply, and after connecting, preservative film and adhesive tape sealing for this hole, guarantees that anesthesia exposes case air tight.Open constant temperature heating plate, 37 DEG C of design temperatures, keep mouse temperature constant in 37 ± 0.5 DEG C.Experiment starts front testing fixture seal, and determining device seal is good.
Induction of anesthesia: with embodiment 1,10 C57BL/6J Mouse Weights are born the 2-3 month between 25g-33g, put into anesthesia and expose case beginning induction of anesthesia, open oxygen tank 1, adjust flux is 2L/min, opens inhalation anesthetic isoflurane volatilization tank 2, regulating concentration is 2.5%, open multiparameter anaesthetic gas monitor 8 simultaneously Exhaust Gas anesthetic concentration is carried out to Real-Time Monitoring, until experiment mice, anyway when areflexia, induction of anesthesia completes.
Anesthesia maintains and presss from both sides tail measuring inhalation anesthetic MAC value: by unified 10 mices be put into lie on one's side be positioned on hot plate, at the edge of hot plate tiling sodica calx for sucking carbon dioxide and the steam of discharge gas.Inhalation anesthetic isoflurane concentration is turned down to 0.7%, and moment observation discharge aspiration enters anesthetic concentration, it is also maintained to 0.7%, after this concentration 30min to be maintained, the tail of every mice is pulled out from animal head fixing hole 14, by centre 1/3 part of 15cm hemostasis clamp mousetail, dynamics is unified to be fixed on first tooth, persistent period 1min for mosquito forceps respectively.Record the mice number to folder tail experiment generation body moving (it is head or the autotelic movement of extremity except breathing that body moves standard), having there is positive reaction to pressing from both sides moving being recorded as of tail experiment generation body in mice, calculates the positive reaction rate of 10 mices.Using the same method and adjusting respectively anesthetic concentration is 0.7%, 0.9%, 1.1%, 1.4%, 1.7%, and the ratio of adjacent two anesthetic concentrations is 0.8, the positive reaction rate of 10 mices while recording each anesthetic concentration respectively.Each adjustment after anesthetic concentration, need to maintain after 30min in this concentration, and with above-mentioned folder tail method repetition aforesaid operations, it is moving whether observation mice body occurs.The position of the each folder of mosquito forceps is the adjacent position at folded position last time, cannot repeat to press from both sides same position, and only in the middle of mice, 1/3 tail place folder is because the middle part of mousetail is the most responsive to pain stimulation.Experiment finishes, and statistical experiment data are used SPSS software, and it is inhalational anesthetic MAC value that the application Return Law is calculated inhalational anesthetic ED50.
This experimental example can prove, apparatus of the present invention also can be for doing the experiment of inhalational anesthetic MAC value experiment and ache related aspect.
embodiment 4a kind ofly apply traditional inhalation anesthesia exposing device and keep adult mice autonomous respiration to keep homoiothermal inhalation anesthesia to expose experiment and and the comparison of experimental provision of the present invention
With reference to accompanying drawing 3, anaesthetize and expose experiment, in Fig. 3, oxygen or empty oxygen gas mixture tank 1, inhalation anesthetic volatilization tank 2, carrier gas pipe 3, air inlet 4, airtight inhalation anesthetic expose the rat/mouse 16, sodica calx 18, the constant water bath box 20 that in case 5, venthole 6, loop, use anesthetic gas absorber 7, multiparameter anaesthetic gas monitor 8, anesthesia to expose.Detailed process is as follows
Before experiment, prepare: with reference to Fig. 3, first open constant water bath box 20 before experiment, design temperature is 37 DEG C, reach after design temperature until water bath temperature, device is connected to testing fixture seal as Fig. 3, guarantee that device seal is good, guarantee that anesthesia exposes the case gas leakage of can not leaking.
Induction of anesthesia: 3 C57BL/6J Mouse Weights are born the 2-3 month between 25g-33g, put into anesthesia and expose case beginning induction of anesthesia, open oxygen tank 1, adjust flux is 2L/min, open inhalation anesthetic isoflurane volatilization tank 2, regulating concentration is 2.5%, opens multiparameter anaesthetic gas monitor 8 simultaneously Exhaust Gas anesthetic concentration is carried out to Real-Time Monitoring, until experiment mice, anyway when areflexia, induction of anesthesia completes.
Anesthesia maintains: after induction of anesthesia finishes, sodica calx 18 is laid in to anesthesia and exposes the interior low side surface of case 5, contribute to absorb mouse breathing and discharge carbon dioxide and the steam in gas.It is 1.4% that inhalation anesthetic isoflurane concentration is turned down, and the moment observe discharge aspiration enter anesthetic concentration, it is also maintained to 1.4%, the waste gas of discharge, by using anesthetic gas absorber 7 to absorb in loop, prevents that waste gas from entering air and polluting.Anaesthetize the 2h that holds time.
Arterial blood gas analysis: maintain 1h, 1.5h, 2h in anesthesia respectively and a mice is wherein extracted to arterial blood carry out arterial blood gas analysis.Detailed process is as follows: rapidly mice is taken out to operating board in the exposure of the anesthesia from water bath 20 case 5 and carry out operation technique.Because mice has departed from inhalation anesthetic exposure, operation stimulation easily impels it to wake up, so operation must be very fast, requires experimental implementation to complete in 1 minute.Mice is fixed on operating board, open rapidly mice thoracic cavity, expose mouse heart, 1 milliliter of syringe of anticoagulant is punctured into left ventricle, visible cherry arterial blood enters in syringe, extract 0.5-1 milliliter arterial blood, to blood gas analyzer (Gem premier 3000), carry out blood gas analysis as early as possible.But owing to not taking Insulation in operating process, mouse breathing frequency obviously reduces, and has occurred obvious respiration inhibition, and the arterial blood gas PH extracting is on the low side, PaCO
2be worth higher.
Done altogether 12 mices with traditional inhalation anesthetic process for exposing, the result of blood gas analysis is referring to table 3.
The mice Results of Blood-gas of comparison sheet 2 and table 3, is not difficult to find to use traditional inhalation anesthetic exposing device to have very large problem, from vim and vigour result, uses the mice pH value compared with normal value of traditional inhalation anesthetic exposing device on the low side, PaCO
2higher.But because mice has stopped inhalational anesthetic exposure in the time operating, in the time extracting the operation of mice left ventricle arterial blood, the actual anaesthetic concentration of mice is lower than exposure value, while stopping the exposure of anesthetics while even exposure as desflurane for the lower inhalation anesthetic of some blood & air partition coefficents, mice can revive rapidly, the stimulation of adding operation technique more easily makes mice revive, the vim and vigour situation when reaction inhalation anesthetic that measured Results of Blood-gas can not be real exposes.And, open thoracic cavity and easily cause the meetings such as pneumothorax affect Results of Blood-gas.Operating aspect: the first, use traditional inhalation anesthetic exposing device carry out blood gas analysis take out arterial blood operation requirements speed must be fast, skilled operation, has increased use difficulty.The second, mouse heart is less, and the position of left ventricle is less, puncture when left ventricle difficulty larger, be especially easily strayed into other atrial ventricle and phlebotomize by mistake, this point operation easier is also larger.Our contrive equipment has greater advantage compared with traditional exposing device, not only easy grasp simple to operate, and measured Results of Blood-gas is also genuine and believable.
Above embodiment can absolutely prove, the device that we invent keeps the inhalation anesthetic of autonomous respiration to expose for adult mice/rat larger advantage, animal heat maintains the monitoring that instrument is sensitiveer and controls animal heat, makes the laboratory animal can be because of hypothermia or too high and occur the abnormal of physical signs in anaesthesia process.The advantage that this device is larger is conveniently to carry out a lot of experimental implementation to the mice in anesthesia process-exposed, as used in embodiment 2, mice is carried out arterial blood gas analysis and has the monitoring of wound arteriotony, thereby and in embodiment 3 animal in anesthesia used carry out pain stimulation and determine inhalational anesthetic MAC value.Our device can also do other a lot of experiment, as the mensuration of the mechanical threshold of pain in mouse anesthesia process-exposed and burning pain threshold, to the administration of anesthesia small mouse, and other more operation techniques etc., convenient and simple, very practical, the application of more devices needs further to be researched and developed.
Claims (3)
1. an inhalation anesthesia exposes and monitoring device, it is characterized in that, this device is by oxygen or empty oxygen gas mixture tank (1), inhalation anesthetic volatilization tank (2), carrier gas pipe (3), air inlet (4), inhalation anesthetic exposes case (5), venthole (6), anesthetic gas absorber (7), multiparameter anaesthetic gas monitor (8), animal heat maintains instrument (9), circuit (10), circuit access hole (11), anatomic microscope (12), constant temperature heating plate (13), mice fixing hole (14), there is wound arteriotony monitor (15), insulation blanket (17), operating board (19) composition, air inlet (4) is opened in the middle of inhalation anesthetic exposes case (5) right side, venthole (6) is opened at inhalation anesthetic and is exposed top, case (5) left side, circuit access hole (11) is opened at inhalation anesthetic and is exposed below, case (5) left side, mice fixing hole (14) is opened at inhalation anesthetic and is exposed the positive below of case (5), inhalation anesthetic exposes case (5) and is placed on operating board (19), oxygen or empty oxygen gas mixture tank (1) connect inhalation anesthetic volatilization tank (2) through carrier gas pipe (3), connect inhalation anesthetic through air inlet (4) again and expose case (5), anesthetic gas absorber (7) is connected inhalation anesthetic with multiparameter anaesthetic gas monitor (8) through venthole (6) and exposes case (5), animal heat maintains instrument (9) and is connected with insulation blanket (17) by circuit (10) via line access hole (11), insulation blanket (17) is placed in inhalation anesthetic and exposes case (5) bottom, mice fixing hole (14) is opened at inhalation anesthetic and is exposed the positive below of case (5), constant temperature heating plate (13) is positioned at mice fixing hole (14) below, anatomic microscope (12) is placed on operating board (19), near constant temperature heating plate (13) and mice fixing hole (14), there is wound arteriotony monitor (15) to be placed on operating board (19).
2. a kind of inhalation anesthesia according to claim 1 exposes and monitoring device, it is characterized in that, there is wound arteriotony monitor (15) to be placed on operating board (19) upper, in the time that needs use has the monitoring of wound arteriotony, be connected with the tremulous pulse of laboratory animal by tremulous pulse pressure-measuring pipe.
3. a kind of inhalation anesthesia according to claim 1 exposes and monitoring device, it is characterized in that, it is airtight that inhalation anesthetic exposes case (5), and material is lucite material, is shaped as cuboid, and its size is suitable mutually with the size of insulation blanket (17).
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| CN108272525A (en) * | 2018-01-12 | 2018-07-13 | 暨南大学 | A kind of anesthesia outfit and the method for preparing artery occlusion model in Balb/c mouse brains |
| CN108670491A (en) * | 2018-06-11 | 2018-10-19 | 新乡医学院 | A kind of mouse cerebrospinal fluid acquisition system and acquisition method |
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| CN108670491A (en) * | 2018-06-11 | 2018-10-19 | 新乡医学院 | A kind of mouse cerebrospinal fluid acquisition system and acquisition method |
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