CN102121917A - Device and method for measuring respiration of microbes - Google Patents
Device and method for measuring respiration of microbes Download PDFInfo
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- CN102121917A CN102121917A CN201110048837XA CN201110048837A CN102121917A CN 102121917 A CN102121917 A CN 102121917A CN 201110048837X A CN201110048837X A CN 201110048837XA CN 201110048837 A CN201110048837 A CN 201110048837A CN 102121917 A CN102121917 A CN 102121917A
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Abstract
The invention provides a device for measuring the respiration of microbes. The device is equipped with a tubular reaction cavity, wherein, the inner wall of the tubular reaction cavity is equipped with a reaction sieve which divides the reaction cavity into an upper part and a lower part; the orifice of the tubular reaction cavity is equipped with a seal plug; the seal plug is equipped with a through hole; an electrode tip hermetically passes through the through hole to extend into the upper part of the tubular reaction cavity; the tubular reaction cavity is vertically placed in a thermostatic water bath, and the thermostatic water bath is equipped with a water inlet and a water outlet which are communicated with a thermostatic water tank for circulation; the bottom surface of the tubular reaction cavity is contacted with the bottom surface of the thermostatic water bath; the top of the thermostatic water bath is equipped with a hole which is hermetically matched with the external diameter of the orifice of the tubular reaction cavity; a magnetic sheet is placed on the bottom surface of the tubular reaction cavity; the bottom surface of the thermostatic water bath is placed on a magnetic stirrer; the tubular reaction cavity is filled with a reaction solution; and a biological sample is placed on the reaction sieve, and the sieve pore is smaller than the volume of the biological sample.
Description
Technical field
The present invention relates to respiratory research of biology and mensuration, relate in particular to the respiratory devices and methods therefor of a kind of mensuration tiny organism researchs such as microorganism in biological membrane, sediment or the water body and small-sized biologies.
Background technology
Surface water body oxygen is to keep biology, as fish, and the pacing items of existence such as zoobenthos, and be most important reactant in the biomass geochemistry circulation.Hydrobiont is an important motivity of water body oxygen expenditure and organic matter degradation to the utilization of water body oxygen; Biological aerobic respiration, decomposes organism exhaustive oxidations such as carbohydrate by the catalytic action of enzyme being meant biosome in the presence of aerobic, produces carbon dioxide and water, discharges the process of big energy simultaneously.Accurately mensuration water body biology is crucial to the speed of utilizing of oxygen to understanding surface water body and sediment-water termination place oxygen power variation.
Biological aerobic respiration is generally represented the consumption of oxygen with biological in the unit interval.Measure biological respiration, the content of oxygen carried out when the content of oxygen and experiment stopped in the time of can adopting the method determination experiment of oxygen electrode or chemistry titration initial; Also can adopt the continuous variation of oxygen in the mode determination experiment process of oxygen electrode to carry out.Along with the universal use of oxygen electrode, biological respiratory mensuration is progressively tended to use oxygen electrode to measure oxygen content.Yet present assay method all is difficult to the mensuration of competent small size bio tank/oxygen consumption.Give birth to biological membrane or stone is given birth to biological membrane such as, leaf, itself contained microbes biomass is less, and the mensuration system of larger volume is difficult to reflect sensitively biomembranous respiration.For another example, when the respiration of the water earthworm or the midge young was measured, the mensuration system of large volume was difficult to equally reflect that small oxygen changes.Therefore, at present on the respiratory assay method of tiny organism, still have bigger vacancy, the simple and practical method of demanding urgently is remedied.
Summary of the invention
The present invention is directed to the deficiency that prior art exists, provide a kind of mensuration tiny organism respiratory little respiratory system, the mensuration system introduces the optics microelectrode that does not consume oxygen, measure the respiration and the photosynthesis of tiny organism, can measure the content of oxygen in the micro updating small samples, satisfy the requirement of measuring small system sample oxygen expenditure speed.
To achieve these goals, the technical solution used in the present invention is: the respiratory device of a kind of mensuration tiny organism, it is characterized in that: a tubular reactor cavity is set, lumen wall is provided with a reaction sieve tubular reactor cavity is separated into two parts up and down, the mouth of pipe is provided with sealing-plug, sealing-plug is provided with a through hole, and this through hole of electrode head hermetically passing stretches into the first half of tubular reactor cavity; The tubular reactor cavity vertically is positioned in the water bath with thermostatic control, water bath with thermostatic control is provided with entery and delivery port and Water Tank with Temp.-controlled communication loop, tubular reactor cavity bottom surface contacts with the water bath with thermostatic control bottom surface, the water bath with thermostatic control top is provided with the hole that the mouth of pipe external diameter with the tubular reactor cavity matches, sealing between the two, place magnetic sheet on the bottom surface of tubular reactor cavity, the water bath with thermostatic control bottom surface places on the magnetic stirring apparatus, fill with reactant liquor in the tubular reactor cavity, biological sample is placed on the reaction sieve, and the reaction sieve aperture is less than the volume of biological sample.
The aperture of described reaction sieve is 20-1000 μ m, and electrode head sieves less than 0.5cm apart from reaction.
Described tubular reactor cavity, reaction sieve, water bath with thermostatic control, sealing-plug is the transparent body.
Use said apparatus and measure the respiratory method of tiny organism, according to the following steps:
(1) with the corresponding connection of intake-outlet of intake-outlet with the constant temperature cavity of Water Tank with Temp.-controlled, open Water Tank with Temp.-controlled, regulate water temperature to default steady temperature, the original position water water temperature of Water Tank with Temp.-controlled water temperature and biological sample sampling site is close, opens water pump and makes Water Tank with Temp.-controlled water outlet and the water bath with thermostatic control of flowing through circulation;
(2) magnet rotor is put into reaction cavity, will react sieve and be placed on the reaction cavity inwall fulcrum;
(3) fill with reactant liquor in reaction cavity, this reactant liquor is the original position water of biological sample sampling site;
(4) place zoobenthos or biological membrane on the reaction sieve, sealing-plug on the froth;
(5) microelectrode is passed silica gel sealing pad and sealing-plug through hole, expose on the reaction sieve of microelectrode head sensitive part in reaction cavity;
(6) start magnetic stirring apparatus, and regulate its frequency modulation motor rotating speed, the rotating speed size is advisable to reach the top water body uniform rotation of reaction sieve;
(7) writing time is opened the dissolved oxygen instrument that is connected with electrode in experiment beginning, record primary oxygen concentration, and the variation of dissolved oxygen concentration in the observing response liquid regularly detects the water body dissolved oxygen concentration;
(8) treat that dissolved oxygen DO significantly is reduced to a numerical value after, obtain the fall off rate of oxygen in this time period; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, closes calibration cell and dissolved oxygen instrument, takes out microelectrode, opens sealing-plug, and reactant liquor and biological sample in the cleaning reaction cavity are measured and finished.
Advantage of the present invention and remarkable result: it has little respiratory system device of little volume reaction cavity, and this little respiratory system device is made simple, needs the sample amount little, and measurement result is stable, applicable to laboratory mostly to respiratory mensuration requirement.For the laboratory of non-commercial use, can be according to the present invention content, process or repack into similar little respiratory system voluntarily.The introducing of microelectrode particularly, for the mensuration of oxygen expenditure speed in the small samples, is a good news.Therefore the responsive head of microelectrode can, utilize the microelectrode technology less than 20 microns, the reaction cavity (2 milliliters of reaction cavity volumes) of design small size, all kinds of biological samples be can place in this small cavity, as biological membrane or the sediment of microorganism contained, small-sized biology (as midge) etc.
Description of drawings
The front elevation of Fig. 1 apparatus of the present invention;
The vertical view of Fig. 2 apparatus of the present invention.
Embodiment
The number in the figure explanation: 1, the microelectrode line connects dissolved oxygen meter; 2, microelectrode; 3, silicagel pad; 4, tube-shaped through hole; 5, sealing-plug; 6, water bath with thermostatic control; 7, reaction cavity; 8, reaction sieve sieve aperture; 9, the reaction sieve; 10, reaction sieve fulcrum; 11, magnet rotor; 12, entery and delivery port; 13, magnet; 14, magnetic stirring apparatus; 15, frequency modulation motor; 16, microelectrode head sensitive part.
Referring to Fig. 1,2, a tubular reactor cavity 7 (4 centimetres of 8 millimeters of calibers, sealing-plug height below 5) is set, and is equipped with sealing system, water bath with thermostatic control system, perturbed system and microelectrode detection system.Middle part tool fulcrum 10 in the reaction cavity 7, fulcrum top placing response sieve 9; The sieve aperture 8 of reaction sieve, reaction cavity 7 and sealing-plug 5 seamless links, sealing-plug center tool one tube orifice 4, tube orifice 4 tops are provided with silica gel sealing pad 3.Reaction cavity is built in the water bath with thermostatic control 6, and the recirculated water in the Water Tank with Temp.-controlled (not shown) is pumped out, and flows through through intake-outlet 12 and returns Water Tank with Temp.-controlled after the water bath with thermostatic control 6.Reaction cavity 7 places on the magnetic stirring apparatus 14; The reaction cavity inner bottom part is provided with a magnet rotor 11.Microelectrode 2 passes silicagel pad 3, exposes the microelectrode sensitive parts in reaction cavity 7 epicoeles after the tube orifice 4 by sealing-plug 5 centers.Wherein, reaction cavity 7, reaction sieve 9, water bath with thermostatic control 6, sealing-plug 5 is transparent quality.
The mensuration process of this device is as follows:
(1) with the corresponding connection of intake-outlet of intake-outlet with the constant temperature cavity of Water Tank with Temp.-controlled, open Water Tank with Temp.-controlled, regulate water temperature to default steady temperature, the original position water water temperature of Water Tank with Temp.-controlled water temperature and biological sample sampling site is close, opens water pump and makes Water Tank with Temp.-controlled water outlet and the water bath with thermostatic control of flowing through circulation;
(2) magnet rotor is put into reaction cavity, will react sieve and be placed on the reaction cavity inwall fulcrum;
(3) fill with reactant liquor in reaction cavity, this reactant liquor is the original position water of biological sample sampling site;
(4) place zoobenthos or biological membrane on the reaction sieve, sealing-plug on the froth;
(5) microelectrode is passed silica gel sealing pad and sealing-plug through hole, expose on the reaction sieve of microelectrode head sensitive part in reaction cavity;
(6) start magnetic stirring apparatus, and regulate its frequency modulation motor rotating speed, the rotating speed size is advisable to reach the top water body uniform rotation of reaction sieve;
(7) writing time is opened the dissolved oxygen instrument that is connected with electrode in experiment beginning, record primary oxygen concentration, and the variation of dissolved oxygen concentration in the observing response liquid regularly detects the water body dissolved oxygen concentration;
(8) treat that dissolved oxygen DO significantly is reduced to a numerical value after, obtain the fall off rate of oxygen in this time period; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, closes calibration cell and dissolved oxygen instrument, takes out microelectrode, opens sealing-plug, and reactant liquor and biological sample in the cleaning reaction cavity are measured and finished.
Assay method implementation process for explanation apparatus of the present invention, we select Taihu Lake Sediment, it is research object that midge in the Taihu Lake Sediment and Yunnan old Jun Mountain streams stone is given birth to biological membrane, just measure the respiratory rate of midge in the Taihu Lake Sediment, Taihu Lake Sediment oxygen consumption rate under the condition of different temperatures, and the living biomembranous clean primary productivity of streams stone/heterotrophism respiratory rate etc. is introduced respectively.Above-mentioned midge and sediment or stone are given birth to the microorganism in the biological membrane, and its individuality is all less, seek out relatively large sample size also relatively more difficult (giving birth to biological membrane as stone), and therefore, the reaction unit of larger volume is difficult to finish default experiment purpose; Select the device of small size, guarantee the temperature stabilization of reaction system in the experimentation and the variation of accurately determining the oxygen content in the small size reaction system, seem particularly important.
1, the respiratory rate of midge in the Taihu Lake Sediment
Gather the Taihu Lake surface deposit in July, 2008, and sieve is got sediment acquisition midge; Gather the overlying water at sediment-water termination place simultaneously.During sampling, the temperature of sediment-water termination is 21 ℃.The present case purpose is to understand the respiratory rate of midge under the in-situ temperature condition, and concrete assay method is as follows:
(1) intake-outlet with calibration cell is connected with the intake-outlet of constant temperature cavity, opens calibration cell, and be adjusted to 21 ℃ of default steady temperatures, and allow water pump go out calibration cell, and the constant temperature cavity of flowing through.The solubleness of gas in water body is comparatively responsive to temperature, and the respiratory rate of temperature and biology is closely related, and the temperature in the therefore strict control experimentation is most important.The constant temperature water temperature can be determined according to the actual temperature in when sampling, also can determine according to corresponding experimental design.In present case, the actual water temperature during sampling is 21 ℃, so in order to obtain the respiratory rate of midge under the physical condition, we select 21 ℃ as experimental temperature.
(2) magnet rotor is put into the reaction cavity bottom, and will be reacted and sieve on the reaction sieve fulcrum that is placed in the reaction cavity.
(3) selection of reaction mesh size, relevant with the individual size of biological sample to be measured.The effect of reaction sieve is to provide a supporting surface, guarantees that testing sample can be positioned in experimentation above the reaction sieve, and the magnet rotor of avoiding reacting the sieve below damages biological sample to be measured.As testing sample in the present case is midge, and its individual lengths can reach one centimetre, and its individual diameter can only be 1000 μ m, and therefore selecting the aperture is the reaction sieve of 500 μ m, can guarantee that generally midge does not fall to reaction sieve below in the experimentation.
(4) in reaction cavity, fill with reactant liquor (being the lake water of 0.2 μ m membrane filtration in the present case); 0.2 μ m filters and can remove most of microorganism and particle in the water body, makes not contain biology or the particulate matter that tool consumes oxygen in the reactant liquor, to reduce or remit in the reactant liquor oxygen expenditure to the influence of midge oxygen expenditure speed as far as possible.
(5) adopt that tweezers are undamaged to be folded up or adopt toothpick gently to choose four of midges to be put on the reaction sieve.Then, sealing-plug on the froth allows part not contain the reactant liquor overflow of target-finding thing.
(6) microelectrode is passed the tube orifice of silicagel pad and sealing-plug, and compress; Silicagel pad is close to the top of tube orifice, can reach the purpose to the reaction cavity sealing like this.The tube orifice of sealing-plug is relevant with the diameter of little electricity level of use.The microelectrode diameter that we use is 1mm, and therefore, the diameter of sealing-plug tube orifice is 1.1mm.
(7) expose the microelectrode sensitive part in reactant liquor, the microelectrode sensitive part is controlled in the 0.5cm scope apart from the distance of reaction sieve, can responsive variation of writing down dissolved oxygen DO to guarantee microelectrode.
(8) start magnetic stirring apparatus, it is 30rmp/min that the adjusting frequency modulation motor makes the rotating speed of magnet rotor; The rotating speed size is advisable to reach the unidirectional uniform rotation of reaction sieve top water body, can guarantee that like this water body of entire reaction cavity is in the mixing state.
(9) open dissolved oxygen instrument, the variation of dissolved oxygen concentration in the observing response liquid, under the normal condition, can there be violent variation in dissolved oxygen concentration.
(10) record primary oxygen concentration 7.2mg/l, water body dissolved oxygen concentration of per 5 seconds records.
(11) behind the 30min, dissolved oxygen DO significantly is reduced to 6.5mg/l, thereby obtains the fall off rate of oxygen in the 15min; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, and the fall off rate of oxygen is near linear in this oxygen reduction scope.
(12) close calibration cell and dissolved oxygen instrument, from the tube orifice of sealing-plug and silicagel pad, take out microelectrode, open sealing-plug, clean out reactant liquor and biological sample midge in the reaction cavity.
(13) experiment finishes.
2, Taihu Lake Sediment oxygen consumption rate under the condition of different temperatures
Gather the Taihu Lake surface deposit in Dec, 2009, gather the overlying water at sediment-water termination place simultaneously.During sampling, the temperature of sediment-water termination is 12 ℃.After the purpose of present case is to measure the temperature increase, the variation of sediment oxygen consumption rate.Concrete mensuration scheme is as follows:
(1) intake-outlet with calibration cell is connected with the intake-outlet of constant temperature cavity, opens calibration cell, and be adjusted to 12 ℃ of default steady temperatures, and allow water pump go out calibration cell, and the constant temperature cavity of flowing through.Solubleness and the temperature of gas in water body is comparatively responsive, and the respiratory rate of temperature and biology is closely related, and the temperature in the therefore strict control experimentation is most important.The constant temperature water temperature can be determined according to the actual temperature in when sampling, also can determine according to corresponding experimental design.
(2) magnet rotor is put into the reaction cavity bottom, and will be reacted and sieve on the reaction sieve fulcrum that is placed in the reaction cavity.
(3) selection of reaction mesh size is relevant with the size of biological sample to be measured.The effect of reaction sieve is to provide a supporting surface, guarantees that testing sample can be positioned in experimentation above the reaction sieve, and the magnet rotor of avoiding reacting the sieve below injures biosome to be measured.Therefore as testing sample in the present case is sediment, selects the reaction sieve of 20 μ m of aperture minimum, can guarantee that like this particle that major part contains the tiny organism body can be suspended in reaction sieve top.
(4) in reaction cavity, fill with the lake water of 0.2 μ m membrane filtration; 0.2 μ m filters and can remove most of microorganism and particle in the water body, does not contain biology or material that tool consumes oxygen in the reactant liquor, to reduce or remit in the reactant liquor oxygen expenditure to the influence of sediment oxygen expenditure speed as far as possible.
(5) adopt 2ml reactant liquor dilution 0.1g sediment, obtain even mixed liquor, the mixed liquor that uses pipettor or micro syringe to draw 0.2ml is then put down gently in reaction sieve top.Then, sealing-plug on the froth allows part not contain sedimental reactant liquor overflow.
(6) tube orifice of sealing-plug is relevant with the diameter of little electricity level of use.The microelectrode diameter that we use is 1mm, and therefore, the diameter of tube orifice is for being defined as 1.1mm.
(7) microelectrode is passed the tube orifice of silicagel pad and sealing-plug, and compress; Silicagel pad is close to the top of tube orifice, can reach the purpose to the reaction cavity sealing like this.
(8) expose the microelectrode sensitive part in reaction cavity, the microelectrode sensitive part is controlled in the 0.5cm scope apart from the distance of reaction sieve, can responsive variation of writing down dissolved oxygen DO to guarantee microelectrode.
(9) start magnetic stirring apparatus, it is 30rmp/min that the adjusting frequency modulation motor makes the rotating speed of magnet rotor; The rotating speed size is advisable to reach the unidirectional uniform rotation of reaction sieve top water body, can guarantee that like this water body of entire reaction cavity is in the mixing state.
(10) open dissolved oxygen instrument, the variation of dissolved oxygen concentration in the observing response liquid, under the normal condition, can there be violent variation in dissolved oxygen concentration.
(11) record primary oxygen concentration is 9.8mg/l, water body dissolved oxygen concentration of per 5 seconds records.
(12) behind the 20min, dissolved oxygen DO significantly is reduced to 9.0mg/l, thereby obtains the fall off rate of oxygen in the certain hour section; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, and the fall off rate of oxygen is near linear in this oxygen reduction scope.
(13) close dissolved oxygen instrument.
(14) regulate calibration cell to 15 ℃ of steady temperatures, keep thermostatted water to be pumped out calibration cell, and the constant temperature cavity of flowing through.Wait for 20min quietly, guarantee to react cavity temperature and reach 15 ℃ of preset temperatures.
(15) repeat the 10-12 step, the dissolved oxygen concentration in the record reaction chamber obtains the speed that oxygen descends in time under 15 ℃ of conditions over time.
(16) close calibration cell and dissolved oxygen instrument, from the tube orifice of sealing-plug and silicagel pad, take out microelectrode, open sealing-plug, clean out reactant liquor and sediment in the reaction cavity.
(17) experiment finishes.
3, streams stone is given birth to the mensuration of biomembranous clean primary productivity/heterotrophism respiratory rate
Gather the stone in old Jun Mountain streams, Yunnan in November, 2009 and give birth to diffraction patterns for biomembrane samples, gather stream simultaneously, take back the laboratory under the deepfreeze condition in the lump.During sampling, the temperature of sediment-water termination is 9 ℃.The present case purpose is to obtain respiratory rate and the photosynthetic rate that streams stone is given birth to diffraction patterns for biomembrane samples, and it is as follows specifically to measure scheme:
(1) intake-outlet with calibration cell is connected with the intake-outlet of constant temperature cavity, opens calibration cell, and be adjusted to 10 ℃ of default steady temperatures, and allow water pump go out calibration cell, and the constant temperature cavity of flowing through.Solubleness and the temperature of gas in water body is comparatively responsive, and the respiratory rate of temperature and biology is closely related, and the temperature in the therefore strict control experimentation is most important.
(2) magnet rotor is put into the reaction cavity bottom, and will be reacted and sieve on the reaction sieve fulcrum that is placed in the reaction cavity.
(3) selection of reaction mesh size is relevant with testing sample.The effect of reaction sieve is to provide a supporting surface, guarantees that testing sample can be positioned in experimentation above the reaction sieve, and the magnet rotor of avoiding reacting the sieve below injures biosome to be measured.As testing sample in the present case is that stone is given birth to biological membrane, and therefore selecting the aperture is the reaction sieve of 20 μ m, can guarantee that like this particle that major part contains the tiny organism body can be suspended in the top of reaction sieve.
(4) in reaction cavity, fill with reactant liquor, as the stream of 0.2 μ m membrane filtration; 0.2 μ m filters and can remove most of microorganism and particle in the stream, does not contain biology or material that tool consumes oxygen in the reactant liquor, to reduce or remit in the reactant liquor oxygen expenditure to the influence of biological membrane oxygen expenditure or generating rate as far as possible.
(5) adopt the rare 0.1g biological membrane of 2ml reactant liquor, obtain even mixed liquor, the mixed liquor that uses pipettor or micro syringe to draw 0.2ml is then put down gently in reaction sieve top.Then, sealing-plug on the froth allows part not contain biomembranous reactant liquor overflow.
(6) tube orifice of sealing-plug is relevant with the diameter of little electricity level of use.The microelectrode diameter that we use is 1mm, and therefore, the diameter of tube orifice is for being defined as 1.1mm.
(7) microelectrode is passed the tube orifice of silicagel pad and sealing-plug, and compress; Silicagel pad is close to the top of tube orifice, can reach the purpose to the reaction cavity sealing like this.
(8) expose the microelectrode sensitive part in reaction cavity, the microelectrode sensitive part is controlled in the 0.5cm scope apart from the distance of reaction sieve, can responsive variation of writing down dissolved oxygen DO to guarantee microelectrode.
(9) start magnetic stirring apparatus, it is 30rmp/min that the adjusting frequency modulation motor makes the rotating speed of magnet rotor; The rotating speed size is advisable to reach the unidirectional uniform rotation of reaction sieve top water body, can guarantee that like this water body of entire reaction cavity is in the mixing state.
(10) cover reaction cavity with light shield, avoid biomembranous photosynthesis.
(11) open dissolved oxygen instrument, the variation of dissolved oxygen concentration in the observing response liquid, under the normal condition, can there be violent variation in dissolved oxygen concentration.
(12) record primary oxygen concentration 9.0mg/l, water body dissolved oxygen concentration of per 5 seconds records.
(13) behind the 15min, dissolved oxygen DO significantly is reduced to 8.3mg/l, thereby obtains the fall off rate of oxygen in the certain hour section, calculates the biomembranous different oxygen respiratory rate of unit mass; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, and the fall off rate of oxygen is near linear in this oxygen reduction scope.
(14) close dissolved oxygen instrument.
(15 remove light shield, for reaction cavity provides light source (60mol/m
2/ s, Metal halogen lamp).
(16) repeat the 11-13 step, the dissolved oxygen concentration in the record reaction chamber obtains the time dependent speed of oxygen over time, calculates the biomembranous hair of unit mass primary productivity.In conjunction with biomembranous different oxygen respiratory rate under the dark condition of the 13rd step acquisition,, calculate biomembranous clean primary productivity according to formula " clean primary productivity=hair primary productivity-heterotrophism respiratory rate ".
(17) close calibration cell and dissolved oxygen instrument, from the tube orifice of sealing-plug and silicagel pad, take out microelectrode, open sealing-plug, clean out reactant liquor and biological membrane in the reaction cavity.Experiment finishes.
Claims (4)
1. measure the respiratory device of tiny organism for one kind, it is characterized in that: a tubular reactor cavity is set, lumen wall is provided with a reaction sieve tubular reactor cavity is separated into two parts up and down, the mouth of pipe is provided with sealing-plug, sealing-plug is provided with a through hole, and this through hole of electrode head hermetically passing stretches into the first half of tubular reactor cavity; The tubular reactor cavity vertically is positioned in the water bath with thermostatic control, water bath with thermostatic control is provided with entery and delivery port and Water Tank with Temp.-controlled communication loop, tubular reactor cavity bottom surface contacts with the water bath with thermostatic control bottom surface, the water bath with thermostatic control top is provided with the hole that the mouth of pipe external diameter with the tubular reactor cavity matches, sealing between the two, place magnetic sheet on the bottom surface of tubular reactor cavity, the water bath with thermostatic control bottom surface places on the magnetic stirring apparatus, fill with reactant liquor in the tubular reactor cavity, biological sample is placed on the reaction sieve, and the reaction sieve aperture is less than the volume of biological sample.
2. the respiratory device of mensuration tiny organism according to claim 1 is characterized in that: the aperture of described reaction sieve is 20-1000 μ m, and electrode head sieves less than 0.5cm apart from reaction.
3. the respiratory device of mensuration tiny organism according to claim 1 and 2 is characterized in that: described tubular reactor cavity, reaction sieve, water bath with thermostatic control, sealing-plug is the transparent body.
4. application rights requires 1 described device to measure the respiratory method of tiny organism, according to the following steps:
(1) with the corresponding connection of intake-outlet of intake-outlet with the constant temperature cavity of Water Tank with Temp.-controlled, open Water Tank with Temp.-controlled, regulate water temperature to default steady temperature, the original position water water temperature of Water Tank with Temp.-controlled water temperature and biological sample sampling site is close, opens water pump and makes Water Tank with Temp.-controlled water outlet and the water bath with thermostatic control of flowing through circulation;
(2) magnet rotor is put into reaction cavity, will react sieve and be placed on the reaction cavity inwall fulcrum;
(3) fill with reactant liquor in reaction cavity, this reactant liquor is the original position water of biological sample sampling site;
(4) place zoobenthos or biological membrane on the reaction sieve, sealing-plug on the froth;
(5) microelectrode is passed silica gel sealing pad and sealing-plug through hole, expose on the reaction sieve of microelectrode head sensitive part in reaction cavity;
(6) start magnetic stirring apparatus, and regulate its frequency modulation motor rotating speed, the rotating speed size is advisable to reach the top water body uniform rotation of reaction sieve;
(7) writing time is opened the dissolved oxygen instrument that is connected with electrode in experiment beginning, record primary oxygen concentration, and the variation of dissolved oxygen concentration in the observing response liquid regularly detects the water body dissolved oxygen concentration;
(8) treat that dissolved oxygen DO significantly is reduced to a numerical value after, obtain the fall off rate of oxygen in this time period; The reduction of dissolved oxygen DO is advisable to be not less than initial oxygen 20%, closes calibration cell and dissolved oxygen instrument, takes out microelectrode, opens sealing-plug, and reactant liquor and biological sample in the cleaning reaction cavity are measured and finished.
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| CN105395202A (en) * | 2015-12-03 | 2016-03-16 | 华东师范大学 | Determination method for basic metabolism condition of aquatic animal |
| CN105395202B (en) * | 2015-12-03 | 2019-03-01 | 华东师范大学 | A kind of measuring method of the basic metabolism situation of aquatic animal |
| CN106018481A (en) * | 2016-05-17 | 2016-10-12 | 中国科学院生态环境研究中心 | Gas-liquid two-phase alternate biological membrane micro-environment detection method |
| CN105928982B (en) * | 2016-05-17 | 2018-07-17 | 中国科学院生态环境研究中心 | A kind of alternate biomembrane microenvironment detection device of gas-liquid two-phase |
| CN106018481B (en) * | 2016-05-17 | 2018-09-21 | 中国科学院生态环境研究中心 | A kind of alternate biomembrane microenvironment detection method of gas-liquid two-phase |
| CN107991471A (en) * | 2017-11-24 | 2018-05-04 | 吉林建筑大学 | A kind of water-biomembrane visible ray shines analogue experiment installation |
| WO2020042211A1 (en) * | 2018-08-28 | 2020-03-05 | 暨南大学 | Respirator bottle for measuring biofilm respiration rate, measurement device and measurement method |
| US11834645B2 (en) | 2018-08-28 | 2023-12-05 | Jinan University | Respirator, measurement device and measuring method for measuring respiratory rate of biofilm |
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