CN103792377A - Device and method for reinforcing smell detection of smell tissue biological sensor by nano zinc - Google Patents
Device and method for reinforcing smell detection of smell tissue biological sensor by nano zinc Download PDFInfo
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- CN103792377A CN103792377A CN201410022421.4A CN201410022421A CN103792377A CN 103792377 A CN103792377 A CN 103792377A CN 201410022421 A CN201410022421 A CN 201410022421A CN 103792377 A CN103792377 A CN 103792377A
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Abstract
The invention discloses a device and a method for reinforcing smell detection of a smell tissue biological sensor by nano zinc. The device introduces nano zinc particles into a smell tissue biological sensor consisting of an olfactory mucosa and a micro electrode array sensor, and meanwhile is combined with an irrigation system, a sensor fixing and temperature preservation system and a data collecting and storage system so as to reinforce a detection signal of the smell tissue biological sensor under smell stimulation. According to the device and method disclosed by the invention, firstly, the spontaneous electric potential of the smell tissue biological sensor is detected to guarantee the activity, and then a nano zinc solution and a smell stimulation solution of to be detected so as to obtain a reinforced detection signal. The device and the method for reinforcing smell detection of the smell tissue biological sensor by nano zinc, which are disclosed by the invention, overcome the defects of low signal to noise ratio and the like of an existing biological sensor smell detection method, and have the advantages of preparation simplicity, convenience in detection, wide application range, high signal to noise ratio and the like, and the device is not required to be fixed; the device and the method can be widely applied to the fields relevant to smell detection, and provide an effectively detection means for the research of a biological sense of smell feeling molecule mechanism.
Description
Technical field
The present invention relates to a kind of odor detection intensifier and method of biology sensor, relate in particular to a kind of Nano-Zinc particle and combine with sense of smell tissue biological sensor and strengthen the device and method that odor sensing detects.
Background technology
Sense of smell tissue biological sensor is to be combined to detect a kind of biology sensor of extraneous Odor stimulation with physics or chemical sensor from the complete olfactory mucosa of peeling off of rat nasal cavity, there is the advantages such as method is simple, biological function reservation is complete, the response time is short, there is significant application value in fields such as biomedicine, environmental monitoring, food industry.But the factors such as the noise that when noise of intrinsic noise, olfactory mucosa and the device coupled interface of senser element and test, external environment interference produces can reduce its signal to noise ratio (S/N ratio), limit its detection to low concentration Odor stimulation, hinder its actual effectively application.In mammalian, contain the Nano-Zinc metallic particles of 1-2nm, the outer electric signal of the born of the same parents that produce when olfactory mucosa is subject to Odor stimulation has humidification.Therefore, the sensing device that Nano-Zinc is combined with sense of smell tissue biological sensor, prepare the bio-sensing detection means simple, easy to detect, signal to noise ratio (S/N ratio) is high as a kind of, for sense of smell tissue biological sensor provides device and method to effective detection and the identification of low concentration Odor stimulation.
Summary of the invention
The object of the invention is to for the deficiencies in the prior art, provide a kind of Nano-Zinc to strengthen the device and method of sense of smell tissue biological sensor odor detection.
The object of the invention is to be achieved through the following technical solutions: a kind of Nano-Zinc strengthens the device of sense of smell tissue biological sensor odor detection, it is made up of perfusion system, biological detection system and data acquisition and storage system three parts, and perfusion system is all connected with biological detection system with data acquisition and storage system; Wherein, perfusion system by oxygen tank, enter oxygen path, sampling system and go out sample system to form, biological detection system is fixed by sense of smell tissue biological sensor, sensor and heat-insulation system and temperature controller form, and data acquisition and storage system are made up of multichannel pmultiple amplifier, data collecting card and computer;
In described perfusion system, sampling system comprises the first sample cell, the second sample cell, the first peristaltic pump, the second peristaltic pump and two-way sample introduction path, goes out sample system and comprises sample path, the 3rd peristaltic pump and waste liquid tank; The input end that enters oxygen path connects oxygen tank, and two output terminals insert respectively the first sample cell and the second sample cell; Two input ends of two-way sample introduction path also insert respectively in the first sample cell and the second sample cell, its output terminal is the test chamber top of adherent insertion sense of smell tissue biological sensor, on two input ends of two-way sample introduction path, the first peristaltic pump and the second peristaltic pump are installed respectively, go out sample path input end the test chamber bottom of the also adherent insertion sense of smell of the 3rd peristaltic pump tissue biological sensor has been installed, output terminal connects waste liquid tank;
In described biological detection system, sense of smell tissue biological sensor is made up of olfactory mucosa and microelectrode array sensor, and sensor is fixed and heat-insulation system is made up of top board and heatable soleplate; Olfactory mucosa cilium towards be above laid in the surface of microelectrode array sensor and with the coupling of microelectrode array sensor generation capacitance-resistance, and microelectrode array sensor used is arranged in 8 × 8 square formations by 64 gold electrodes and forms; Utilize sensor to fix and the top board of heat-insulation system is fixed on sense of smell tissue biological sensor in heatable soleplate, and be used in conjunction with transmission of signal by the phase of solder joint on metal probe and microelectrode array sensor on top board, heatable soleplate connects temperature controller outward;
In described data acquisition and storage system, multichannel pmultiple amplifier and sense of smell tissus sensor are altogether, and the data input pin connecting sensor of multichannel pmultiple amplifier is fixed and the data acquisition serial ports of the top board of heat-insulation system, data output end connection data capture card, data collecting card is connected with computer by USB serial ports again.
Application said apparatus carries out the method for Nano-Zinc enhancing sense of smell tissue biological sensor odor detection, comprises the steps:
(1) obtain the signalling certainly of sense of smell tissue biological sensor: temperature controller temperature is arranged on to 37 ℃, in the first sample cell, add PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min, opens the first peristaltic pump after logical oxygen 5s, perfusion speed 10 μ L/min, after perfusion 2s, open the 3rd peristaltic pump, after perfusion 150s, close the first peristaltic pump, Liquid Residue in emptying test chamber, then close the 3rd peristaltic pump, stop logical oxygen;
(2) application apparatus of the present invention strengthen the detection to Odor stimulation material to be measured: the PBS damping fluid in the first sample cell is replaced with to Odor stimulation substance solution to be measured, by 10
-15m ~ 10
-7the Nano-Zinc solution of M adds the second sample cell in batches, then test respectively according to the following steps: start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump and the second peristaltic pump simultaneously, perfusion speed 10 μ L/min, open the 3rd peristaltic pump after perfusion 1s, close the first peristaltic pump and the second peristaltic pump after perfusion 150s, in chamber to be tested, after residual solution emptying, close the 3rd peristaltic pump, stop logical oxygen; Under the effect of Nano-Zinc, on the olfactory mucosa of sense of smell tissue biological sensor, olfactory receptor cell is subject to the outer electric signal of born of the same parents that Odor stimulation produces and spreads out of by the gold electrode on microelectrode array sensor that sensor is fixed and the metal probe of the top board of heat-insulation system, be transmitted to again top board data acquisition serial ports, and be input to multichannel pmultiple amplifier and carry out the preliminary amplification of data, signal after amplification imports in data collecting card analog electrical signal is converted to digital electric signal again, is finally transmitted in computer and shows;
Sample in the first sample cell is replaced with to PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min, after logical oxygen 5s, open the first peristaltic pump, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 2s, after perfusion 10s, close the first peristaltic pump and the 3rd peristaltic pump, then stop logical oxygen.Active and stand-by to maintain sense of smell tissue biological sensor.
The invention has the beneficial effects as follows, provide a kind of Nano-Zinc to strengthen the device of the odor detection of sense of smell tissue biological sensor, realized the effective detection to Odor stimulation.The present invention has overcome the too low deficiency that waits of existing biology sensor odor detection method signal to noise ratio (S/N ratio), there is simple, easy to detect, applied widely, the signal to noise ratio (S/N ratio) advantages of higher of preparation, and without fixing, to strengthen effect adjustable flexibly, can be widely used in the association area of odor detection, the research of simultaneously also experiencing molecular mechanism for Biologic Olfaction provides effective detection means.
Accompanying drawing explanation
Fig. 1 is the composition schematic diagram that Nano-Zinc of the present invention strengthens sense of smell tissue biological sensor device for testing smell;
Fig. 2 is the sense of smell tissue biological sensor composition of apparatus of the present invention and detects principle schematic;
Fig. 3 be variable concentrations Nano-Zinc of the present invention strengthen sense of smell tissue biological sensor odor detection device to 10
-5the detection method of M isoamyl acetate Odor stimulation and detection signal schematic diagram;
Fig. 4 is the present invention 10
-15the device of M Nano-Zinc enhancing sense of smell tissue biological sensor odor detection and sense of smell tissue biological sensor are respectively to 10
-7m, 10
-3m, 10
-5the detection signal schematic diagram of M isoamyl acetate Odor stimulation;
Fig. 5 is the present invention 10
-15the device of M Nano-Zinc enhancing sense of smell tissue biological sensor odor detection and sense of smell tissue biological sensor are respectively to 10
-7m, 10
-3m, 10
-5the detection signal contrast statistical graph of M isoamyl acetate Odor stimulation;
Fig. 6 is the present invention 10
-15the device of M Nano-Zinc enhancing sense of smell tissue biological sensor odor detection and sense of smell tissue biological sensor are to 10
-7m, 10
-3m, 10
-5the detection signal trend map of M isoamyl acetate Odor stimulation;
In figure: oxygen tank 1, enter oxygen path 2, sampling system 3, the first sample cell 31, the second sample cell 32, the first peristaltic pump 33, the second peristaltic pump 34, two-way sample introduction path 35, go out sample system 4, go out sample path 41, the 3rd peristaltic pump 42, waste liquid tank 43, sense of smell tissue biological sensor 5, olfactory mucosa 51, olfactory receptor cell 52, microelectrode array sensor 53, gold electrode 54, sensor is fixed and heat-insulation system 6, metal probe 61, heatable soleplate 62, temperature-controlling system 7, multichannel amplifier system 8, data collecting card 9, computer 10.
Embodiment
Below in conjunction with drawings and the specific embodiments, the present invention is described in detail, but be not restriction the present invention.
Be illustrated in figure 1 the device design that Nano-Zinc of the present invention strengthens sense of smell tissue biological sensor odor detection, whole device is made up of perfusion system, biological detection system and data acquisition and storage system three parts, and perfusion system is all connected with biological detection system with data acquisition and storage system.Wherein, perfusion system by oxygen tank 1, enter oxygen path 2, sampling system 3 and go out sample system 4 to form; Biological detection system is fixed by sense of smell tissue biological sensor 5, sensor and heat-insulation system 6 and temperature controller 7 form; Data acquisition and storage system are made up of multichannel pmultiple amplifier 8, data collecting card 9 and computer 10.
In perfusion system, sampling system 3 comprises the first sample cell 31, the second sample cell 32, the first peristaltic pump 33, the second peristaltic pump 34 and two-way sample introduction path 35; Comprise sample path 41, the 3rd peristaltic pump 42 and waste liquid tank 43 and go out sample system 4.The input end that enters oxygen path 2 connects 1, two output terminal of oxygen tank and inserts respectively the first sample cell 31 and the second sample cell 32.Simultaneously, in sampling system 3, two input ends of two-way sample introduction path 35 also insert respectively in the first sample cell 31 and the second sample cell 32, its output terminal is the test chamber top of adherent insertion sense of smell tissue biological sensor 5, and the first peristaltic pump 33 and the second peristaltic pump 34 have been installed respectively on two input ends of two-way sample introduction path 35.On the other hand, go out the test chamber bottom that outes sample path 41 input ends the 3rd peristaltic pump 42 adherent insertion sense of smell tissue biological sensor 5 have been installed of sample system 4, output terminal connects waste liquid tank 43.
In biological detection system, sense of smell tissue biological sensor 5 is made up of olfactory mucosa 51 and microelectrode array sensor 53, as shown in Figure 2; And sensor is fixed and heat-insulation system 6 is made up of top board 61 and heatable soleplate 63.The adult SD rats of 200g-300g is implemented to intraperitoneal anesthesia (20% urethane till death, 4mL/100g), ophthalmologic operation is cut and in 75% ethanol, after cleaning and sterilizing, is cut off SD rat nasal bone, open nasal septum and take off nasal septum osteocomma, re-use ophthalmology pincers and put down and scrape the olfactory mucosa 51 being attached on osteocomma by its complete peeling off gently.The olfactory mucosa 51 of peeling off is at the PBS damping fluid (pH=7.2 of 37 ℃, 0.1M refers to phosphatic volumetric molar concentration in PBS damping fluid, the PBS damping fluid using in this patent all refers to 0.1M, the PBS damping fluid of pH=7.2) in rinse and wash twice gently, then by olfactory mucosa 51 ciliums towards the surface that is above laid in microelectrode array sensor 53.Microelectrode array sensor 53 adopts standard miromaching to make, and the diameter of the gold electrode 54 distributing on it is 30 μ m, electrode separation 100 μ m, and 64 gold electrodes 54 are arranged in 8 × 8 square formations.Utilize sensor to fix and the top board 61 of heat-insulation system 6 is fixed on sense of smell tissue biological sensor 5 in heatable soleplate 63, and be connected with the solder joint on microelectrode array sensor 53 with conducting electrical signals to data acquisition serial ports by the metal probe 62 on top board 61, the outer company of heatable soleplate 63 temperature controller 7.Fix in backward sense of smell tissue biological sensor 5 perfusion PBS damping fluid and detach fast, leaving standstill and make olfactory mucosa 51 and microelectrode array sensor 53 that capacitance-resistance coupling occur, completing the assembling of sense of smell tissue biological sensor with fixing.
In data acquisition and storage system, multichannel pmultiple amplifier 8 and sense of smell tissus sensor 5 are altogether, and the data input pin connecting sensor of multichannel pmultiple amplifier 8 is fixed and the data acquisition serial ports of the top board 61 of heat-insulation system 6, data output end connection data capture card 9, data collecting card 9 is connected with computer 10 by USB serial ports again.
After above whole system installation, be placed in shielded box and test, testing procedure is as follows:
1, test the activity of the olfactory mucosa 51 on sense of smell tissue biological sensor 5 and the degree of coupling with microelectrode array sensor 53 thereof.After whole system has connected, connect all power supplys, turn on sensor is fixed and heat-insulation system 6, temperature controller 7, multichannel pmultiple amplifier 8, data collecting card 9 and computer 10.Temperature controller temperature is arranged on to 37 ℃.In the first sample cell 31, add PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 2s, observe the signal detecting.
Olfactory mucosa has biologically active, in the PBS of implantation environment damping fluid, can produce from signaling, if this signal can be detected by sense of smell tissue biological sensor 5, as shown in signal segment while not adding stimulation as Fig. 3, prove that olfactory mucosa 51 has activity good with microelectrode array sensor 53 couplings, also proves that olfactory sensor 5 can effectively detect Odor stimulation.
2, the impact of test Nano-Zinc itself on sense of smell tissue biological sensor 5 detection signals.After proving that olfactory sensor 5 can effectively detect Odor stimulation, close the first peristaltic pump 33, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42, then stop logical oxygen.Then in the second sample cell 32, add 10
-15m ~ 10
-7the Nano-Zinc solution of M, starts logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the second peristaltic pump 34, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 2s, observe the signal detecting.
Close the second peristaltic pump 34, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42, keep logical oxygen, oxygen-supply speed 0.5-1L/min.Open the first peristaltic pump 33, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 1s, close the first peristaltic pump 33 and the 3rd peristaltic pump 42 after perfusion 10s, then stop logical oxygen, active and stand-by to maintain sense of smell tissue biological sensor.
The principle that Nano-Zinc strengthens sense of smell tissus sensor is as follows: 10
-15m ~ 10
-7when M Nano-Zinc individualism, olfactory mucosa 51 is had no stimulation, the signal detecting does not strengthen; And Nano-Zinc particle and scent molecule are while jointly existing, can its response that scent molecule is stimulated be increased the olfactory receptor cell 52 of acting in conjunction in olfactory mucosa 51, olfactory receptor cell 52 cilium ends contact with scent molecule and Nano-Zinc particle, gold electrode 54 on cell cell space end in contact microelectrode array sensor 53, the electrical signal conduction of generation is entered to microelectrode array sensor 53, therefore the increase of response signal can cause detection signal to increase thereupon, thereby realizes the enhancing to sense of smell tissue biological sensor 5 odor detection.As shown in Figure 3, add 10
-15detection signal after M Nano-Zinc solution is as good as with the signalling certainly detecting, proves that this concentration Nano-Zinc itself has no stimulation for olfactory mucosa 51, can be used for the operation of follow-up enhancing odor detection.
Application said system detection of platform Odor stimulation material the method that strengthens detection signal comprise the following steps:
1, obtain the signalling certainly of sense of smell tissue biological sensor 5.
Complete after building of apparatus of the present invention by Fig. 1 and Fig. 2, connect all power supplys, turn on sensor is fixed and heat-insulation system 6, temperature controller 7, multichannel pmultiple amplifier 8, data collecting card 9 and computer 10.Temperature controller 7 temperature are arranged on to 37 ℃, in the first sample cell 31, add PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min, opens the first peristaltic pump 33 after logical oxygen 5s, perfusion speed 10 μ L/min, after perfusion 2s, open the 3rd peristaltic pump 42, after perfusion 150s, close the first peristaltic pump 33, Liquid Residue in emptying test chamber, then close the 3rd peristaltic pump 42, stop logical oxygen.The signal that in test process, real-time monitored produces.
Logical oxygen guarantees that the cell of olfactory mucosa 51 obtains enough oxygen, maintains its physiologically active; Perfusate is as far as possible few, to maintain the good coupling of olfactory mucosa 51 and microelectrode array sensor 53.PBS damping fluid is as the approximate simulation solution of extracellular fluid environment, can make olfactory mucosa produce spontaneous potential, the current potential producing is by the gold electrode 54 on microelectrode array sensor 53, conduct to data acquisition serial ports by the metal probe 62 that contacts the top board 61 being connected with gold electrode 54 again, then be input to the preliminary amplification that multichannel pmultiple amplifier 8 carries out data, signal after amplification imports in data collecting card 9 again analog electrical signal is converted to digital electric signal, is finally transmitted in computer 10 and is shown by USB serial ports.
2, obtain the detection signal of sense of smell tissue biological sensor 5 to Odor stimulation material to be measured.
PBS damping fluid in the first sample cell 31 is replaced with to Odor stimulation substance solution to be measured, in the second sample cell, add PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33 and the second peristaltic pump 34, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 1s, the response signal of Real Time Observation sense of smell tissue biological sensor 5.After test finishes, close the first peristaltic pump 33 and the second peristaltic pump 34, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42, then stop logical oxygen.(as only needed the odor detection signal after Nano-Zinc of the present invention strengthens, this step can be omitted)
3, application system platform of the present invention strengthens the detection of sense of smell tissue biological sensor 5 to Odor stimulation material.
With 10
-15m ~ 10
-7the Nano-Zinc solution of M is replaced solution in the second sample cell 32 successively, then test respectively according to the following steps: start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33 and the second peristaltic pump 34 simultaneously, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 1s, close the first peristaltic pump 33 and the second peristaltic pump 34 after perfusion 150s, in chamber to be tested, after residual solution emptying, close the 3rd peristaltic pump 42, stop logical oxygen.Real Time Observation record the detection signal of sense of smell tissue biological sensor 5 in test process.
Under the effect of Nano-Zinc, on the olfactory mucosa 51 of sense of smell tissue biological sensor 5, olfactory receptor cell 52 is subject to the outer electric signal of born of the same parents that Odor stimulation produces and spreads out of by the gold electrode 54 on microelectrode array sensor 53 that sensor is fixed and the data acquisition serial ports of the top board of heat-insulation system 6 61, and be input to multichannel pmultiple amplifier 8 and carry out data amplification, enlargement factor is 1000, signal after amplification imports in data collecting card 9 again analog electrical signal is converted to digital electric signal, is finally transmitted to and in computer 10, shows and store.Shown in Fig. 3, be 10
-15m ~ 10
-7the sense of smell tissue biological sensor that the Nano-Zinc solution of M strengthens is to 10
-5the detection signal of M isoamyl acetate.
4, the cleaning of apparatus of the present invention and for subsequent use.
Clean and dry up the first sample cell 31 and the second sample cell 32 with nitrogen, then in the first sample cell 31, adding PBS damping fluid, starting logical oxygen, oxygen-supply speed 0.5-1L/min.Open the first peristaltic pump 33, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 1s, close the first peristaltic pump 33 and the 3rd peristaltic pump 42 after perfusion 10s, then stop logical oxygen, active and stand-by to maintain sense of smell tissue biological sensor.
Embodiment
10
-15m Nano-Zinc strengthens sense of smell tissue biological sensor to 10
-7m, 10
-5m, 10
-3the detection method of M isoamyl acetate Odor stimulation, comprises the steps:
1, adopting isoamyl acetate to analyze pure (99%) dilution configuration concentration gradient is 10
-7m, 10
-5m and 10
-3the each 100mL of isoamyl acetate standard model solution of M, configuration concentration is 10 simultaneously
-15the Nano-Zinc solution of M.Solvent is deionized water.All vessel are clean by pure water rinsing, and nitrogen dries up rear use.
2, put up after apparatus of the present invention by Fig. 1 and Fig. 2, connect all power supplys, turn on sensor is fixed and heat-insulation system 6, temperature controller 7, multichannel pmultiple amplifier 8, data collecting card 9 and computer 10.Temperature controller 7 temperature are arranged on to 37 ℃.In the first sample cell 31, add PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 2s, detect and record the signalling certainly of sense of smell tissue biological sensor 5.
3, close the first peristaltic pump 33, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42, then stop logical oxygen.Then in the second sample cell 32, add 10
-15m Nano-Zinc solution, starts logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the second peristaltic pump 34, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 2s, detect signal when also minute book contrive equipment does not add smell excitor substance.
4, close the second peristaltic pump 34, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42.Keep logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 2s, and then perfusion 150s closes the first peristaltic pump 33, closes the 3rd peristaltic pump 42 in chamber to be tested after residual solution emptying, then stop logical oxygen.
Solution in the first sample cell 31 is replaced with to 10 successively
-7m, 10
-5m and 10
-3m isoamyl acetate solution, and the solution in the second sample cell 32 is replaced with to PBS damping fluid, be then repeated below step: start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33 and the second peristaltic pump 34 simultaneously, perfusion speed 10 μ L/min, Hemoperfusion time 150s, opens the 3rd peristaltic pump 42 after perfusion 1s, observe and record detection signal.
5, close the first peristaltic pump 33 and the second peristaltic pump 34, will in sense of smell tissue biological sensor 5 test chambers, after residual solution emptying, close the 3rd peristaltic pump 42, then stop logical oxygen.Solution in the first sample cell 31 is replaced with to 10 successively
-7m, 10
-5m and 10
-3m isoamyl acetate solution, replaces with 10 by the solution in the second sample cell 32
-15m Nano-Zinc solution, and repeat following steps: start logical oxygen, oxygen-supply speed 0.5-1L/min.After logical oxygen 5s, open the first peristaltic pump 33 and the second peristaltic pump 34 simultaneously, perfusion speed 10 μ L/min, after perfusion 1s, open the 3rd peristaltic pump 42, after perfusion 150s, close the first peristaltic pump 33 and the second peristaltic pump 34, in chamber to be tested, after residual solution emptying, close the 3rd peristaltic pump 42.Observe and record detection signal.The interior sample of the first sample cell 31 is replaced with to PBS damping fluid, open the first peristaltic pump 33, perfusion speed 10 μ L/min, open the 3rd peristaltic pump 42 after perfusion 1s, close the first peristaltic pump 33 and the 3rd peristaltic pump 42 after perfusion 10s, then stop logical oxygen.
6, shown in Fig. 4, be the signal that above each step detects, can see that Nano-Zinc does not affect from signaling olfactory mucosa, be that Nano-Zinc itself has no stimulation to sense of smell tissue biological sensor, and in the situation that Odor stimulation exists, Nano-Zinc can effectively strengthen the detection signal of sensor.In order more clearly to explain this enhancing effect, can extract signal peak (threshold value is decided to be 3 times of standard deviations), the peak-to-peak value of the signal that obtains detecting by threshold value extraction method to the signal detecting.Then take the concentration (M) of isoamyl acetate as horizontal ordinate, peak-to-peak value (μ V) is made histogram for ordinate, sees Fig. 5.Known the present invention 10
-15m Nano-Zinc strengthens sense of smell tissus sensor odor detection system to 10
-7m, 10
-5m and 10
-3the isoamyl acetate Odor stimulation of M detects, and the amplitude of the response signal obtaining has remarkable enhancing effect.Fig. 6 according to the peak-to-peak value that detects calculate Nano-Zinc strengthen before and after system of the present invention detect the sensitivity of isoamyl acetate Odor stimulation, sensitivity approximately rises to original 1.58 times, the corresponding lower limit that detects reduces approximately to original 63.36%.
Claims (2)
1. the device of a Nano-Zinc enhancing sense of smell tissue biological sensor odor detection, it is characterized in that, it is made up of perfusion system, biological detection system and data acquisition and storage system three parts, and perfusion system is all connected with biological detection system with data acquisition and storage system; Wherein, perfusion system by oxygen tank (1), enter oxygen path (2), sampling system (3) and go out sample system (4) to form, biological detection system by sense of smell tissue biological sensor (5), sensor fixes and heat-insulation system (6) and temperature controller (7) form, data acquisition and storage system are made up of multichannel pmultiple amplifier (8), data collecting card (9) and computer (10);
In described perfusion system, sampling system (3) comprises the first sample cell (31), the second sample cell (32), the first peristaltic pump (33), the second peristaltic pump (34) and two-way sample introduction path (35), goes out sample system (4) and comprises sample path (41), the 3rd peristaltic pump (42) and waste liquid tank (43); The input end that enters oxygen path (2) connects oxygen tank (1), and two output terminals insert respectively the first sample cell (31) and the second sample cell (32); Two input ends of two-way sample introduction path (35) also insert respectively in the first sample cell (31) and the second sample cell (32), its output terminal is the test chamber top of adherent insertion sense of smell tissue biological's sensor (5), the first peristaltic pump (33) and the second peristaltic pump (34) have been installed respectively on two input ends of two-way sample introduction path (35), go out sample path (41) input end the test chamber bottom of the 3rd peristaltic pump (42) adherent insertion sense of smell tissue biological's sensor (5) has been installed, output terminal connects waste liquid tank (43);
In described biological detection system, sense of smell tissue biological sensor (5) is made up of olfactory mucosa (51) and microelectrode array sensor (53), and sensor fixes and heat-insulation system (6) is made up of top board (61) and heatable soleplate (63); Olfactory mucosa (51) cilium is towards being above laid in the surface of microelectrode array sensor (53) and with microelectrode array sensor (53), capacitance-resistance coupling occurring, and microelectrode array sensor used (53) is arranged in 8 × 8 square formations by 64 gold electrodes (54) and forms; Utilize sensor to fix and the top board (61) of heat-insulation system (6) is fixed on sense of smell tissue biological sensor (5) in heatable soleplate (63), and be used in conjunction with transmission of signal the outer temperature controller (7) that connects of heatable soleplate (63) by the metal probe (62) on top board (61) and the phase of solder joint on microelectrode array sensor (53);
In described data acquisition and storage system, multichannel pmultiple amplifier (8) and sense of smell tissus sensor (5) are altogether, and the data input pin connecting sensor of multichannel pmultiple amplifier (8) is fixed and the data acquisition serial ports of the top board (61) of heat-insulation system (6), data output end connection data capture card (9), data collecting card (9) is connected with computer (10) by USB serial ports again.
2. apply the method that said apparatus carries out Nano-Zinc enhancing sense of smell tissue biological sensor odor detection, comprise the steps:
1. obtain the signalling certainly of sense of smell tissue biological sensor (5): temperature controller (7) temperature is arranged on to 37 ℃, the first sample cell adds PBS damping fluid in (31), start logical oxygen, oxygen-supply speed 0.5-1L/min, opens the first peristaltic pump (33) after logical oxygen 5s, perfusion speed 10 μ L/min, after perfusion 2s, open the 3rd peristaltic pump (42), after perfusion 150s, close the first peristaltic pump (33), Liquid Residue in emptying test chamber, then close the 3rd peristaltic pump (42), stop logical oxygen;
2. apply apparatus of the present invention and strengthen the detection to Odor stimulation material to be measured: the PBS damping fluid in the first sample cell (31) is replaced with to Odor stimulation substance solution to be measured, by 10
-15m ~ 10
-7the Nano-Zinc solution of M adds the second sample cell (32) in batches, then test respectively according to the following steps: start logical oxygen, oxygen-supply speed 0.5-1L/min, after logical oxygen 5s, open the first peristaltic pump (33) and the second peristaltic pump (34) simultaneously, perfusion speed 10 μ L/min, open the 3rd peristaltic pump (42) after perfusion 1s, close the first peristaltic pump (33) and the second peristaltic pump (34) after perfusion 150s, in chamber to be tested, after residual solution emptying, close the 3rd peristaltic pump (42), stop logical oxygen, under the effect of Nano-Zinc, the upper olfactory receptor cell (52) of the olfactory mucosa (51) of sense of smell tissue biological sensor (5) is subject to the outer electric signal of born of the same parents that Odor stimulation produces and spreads out of by the gold electrode (54) on microelectrode array sensor (53) that sensor is fixed and the metal probe of the top board of heat-insulation system (6) (61) (62), be transmitted to again top board (61) data acquisition serial ports, and be input to multichannel pmultiple amplifier (8) and carry out the preliminary amplification of data, signal after amplification imports in data collecting card (9) again analog electrical signal is converted to digital electric signal, finally be transmitted in computer (10) and show, the interior sample of the first sample cell (31) is replaced with to PBS damping fluid, start logical oxygen, oxygen-supply speed 0.5-1L/min, after logical oxygen 5s, open the first peristaltic pump (33), perfusion speed 10 μ L/min, open the 3rd peristaltic pump (42) after perfusion 2s, close the first peristaltic pump (33) and the 3rd peristaltic pump (42) after perfusion 10s, stop again logical oxygen, active and stand-by to maintain sense of smell tissue biological sensor.
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