CN105466979B - Plant organic molecule on-line measuring device and method - Google Patents
Plant organic molecule on-line measuring device and method Download PDFInfo
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Abstract
A kind of method for carrying out organic molecule on-line checking the present invention provides plant organic molecule on-line measuring device and using above-mentioned apparatus.Above-mentioned apparatus includes microbiosensor unit, pumping signal generating unit, data acquisition process unit and main control unit.Main control unit generates original excitation signal and gives pumping signal generating unit, it is set to generate pumping signal, microbiosensor unit generates electric signal corresponding with organic molecule concentration to be detected under the action of pumping signal, electric signal is pre-processed by data acquisition process unit, pretreated electric signal is analyzed by main control unit to obtain organic molecule concentration to be detected later.Microbiosensor unit is inserted into plant organ to be detected when use and is detected, it realizes to the online of plant organic molecule, live body, dynamic detection and continuous analysis, new solution is provided for plant physiology, plant electrophysiology, botany, plant cell biology etc..
Description
Technical field
The invention belongs to monitoring organic compounds technical fields, are more particularly to a kind of plant organic molecule on-line measuring device
And method.
Background technology
Plant organic molecule plays an important roll in growth and development of plants, existing plant organic molecule detection device
Including amino-acid analyzer, microplate reader, gas chromatography-mass spectrometry analysis instrument, efficient liquid phase chromatographic analysis instrument etc..These detection devices
It is detected using analytical chemistry methods, detection process needs in vitro analysis, and is sampled provided with different time points, Zhi Houtong
The pretreatment process such as grinding, homogenate, filtering are crossed, finally upper machine analyzes organic molecule content.
The detection of existing detection technique is the several time point static state concentration of plant organic molecule some or certain, need from
Physical examination is surveyed, and is only capable of obtaining the static information of organic molecule in plant, can not be embodied dynamic response of the plant to environmental change
Mechanism, and detect sample preparation and pretreatment process it is complicated, efficiency is low.
Invention content
The technical problem to be solved by the present invention is to how to realize, live body, dynamic detection online to plant organic molecule with
And analysis.
In order to solve the above technical problem, the present invention provides a kind of plant organic molecule on-line measuring device, the dresses
It sets including microbiosensor unit, pumping signal generating unit, data acquisition process unit and main control unit；
The main control unit is connect with the pumping signal generating unit, and the main control unit generates original excitation signal simultaneously
It is transferred to the pumping signal generating unit；
The pumping signal generating unit is connect with the microbiosensor unit, the pumping signal generating unit
Pumping signal is generated according to the original excitation signal and passes to the microbiosensor unit；
The microbiosensor unit is connect with the data acquisition process unit, the microbiosensor list
Member reacts under the action of the pumping signal with organic molecule to be detected, generates electric signal, and by the electricity of generation
Signal passes to the data acquisition process unit；
The data acquisition process unit is connect with the main control unit, and the data acquisition process unit is to the telecommunications
The main control unit, the pre- place that the main control unit transmits the data acquisition process unit are passed to after number being pre-processed
Electric signal after reason is analyzed to obtain the concentration of organic molecule to be detected.
Preferably, described device further includes pressure drop temperature compensation unit；The pressure drop temperature compensation unit with it is described miniature
Biosensor unit and main control unit connection, the pressure drop temperature compensation unit acquire the microbiosensor
Pressure drop during cell operation and temperature, and pass to the main control unit；
The main control unit generates temperature-compensating excitation letter according to the temperature information that the pressure drop temperature compensation unit transmits
Number and pass to the pumping signal generating unit, the pumping signal generating unit according to the temperature-compensating pumping signal with
And the original excitation signal generates the pumping signal；
The main control unit exports corresponding compensation according to the pressure drop information that the pressure drop temperature compensation unit transmits and compensates
Voltage give the data acquisition process unit, the data acquisition process unit according to the offset voltage to the electric signal into
Row pretreatment, by treated, electric signal passes to the main control unit later.
Preferably, the data acquisition process unit includes signal amplification subelement and signal condition subelement；It is described
Signal amplification subelement is connect with the microbiosensor unit, be used for by the electric signal and the offset voltage into
The pretreatment of row superposition and amplification；The signal condition subelement and signal amplification subelement and the main control unit
Connection, for improving amplified electric signal, passes to the main control unit later.
Preferably, the main control unit includes the first digital-to-analogue conversion subunit and the first analog-to-digital conversion subelement；
The original excitation signal is described sharp by being passed to after first digital-to-analogue conversion subunit progress digital-to-analogue conversion
Encourage signal generating unit；
The pretreated electric signal of data acquisition process unit carries out mould by the first analog-to-digital conversion subelement
The main control unit is passed to after number conversion.
Preferably, the main control unit further includes the second digital-to-analogue conversion subunit, third D/A conversion unit and second
Analog-to-digital conversion subelement；
The temperature-compensating pumping signal passes to institute after carrying out digital-to-analogue conversion by second digital-to-analogue conversion subunit
State pumping signal generating unit；
The pressure drop of the pressure drop temperature compensation unit acquisition and temperature are carried out by the second analog-to-digital conversion subelement
The main control unit is passed to after analog-to-digital conversion；
The offset voltage passes to the miniature life after carrying out digital-to-analogue conversion by the third digital-to-analogue conversion subunit
Object sensor unit.
Preferably, the main control unit further includes master control subelement, and working sensor is stored in the master control subelement
Curve determines the concentration of organic molecule to be detected using the working sensor curve and the electric signal.
Preferably, the electric signal is electric current.
A method of it carrying out plant organic molecule using above-mentioned device and examines online, which is characterized in that the method
Include the following steps：
S1, main control unit generate original excitation signal and are transferred to pumping signal generating unit；
S2, the pumping signal generating unit generate pumping signal according to the original excitation signal and pass to miniature life
Object sensor unit；
S3, the microbiosensor unit are sent out under the action of the pumping signal with organic molecule to be detected
Raw reaction, generates electric signal, and the electric signal of generation is passed to data acquisition process unit；
S4, the data acquisition process unit pass to the main control unit, institute after being pre-processed to the electric signal
State the pretreated electric signal that main control unit transmits the data acquisition process unit analyzed to obtain it is to be detected organic
The concentration of small molecule.
Preferably, the method is further comprising the steps of：
Pressure drop temperature compensation unit acquires pressure drop and temperature during the microbiosensor cell operation, and
Pass to the main control unit；
The main control unit generates temperature-compensating excitation letter according to the temperature information that the pressure drop temperature compensation unit transmits
Number and pass to the pumping signal generating unit, the pumping signal generating unit according to the temperature-compensating pumping signal with
And the original excitation signal generates the pumping signal；
The main control unit exports corresponding offset voltage according to the pressure drop information that the pressure drop temperature compensation unit transmits
To the data acquisition process unit, the data acquisition process unit carries out the electric signal according to the offset voltage pre-
Processing, by treated, electric signal passes to the main control unit later.
Preferably, in the step S4, after the pretreatment that the main control unit transmits the data acquisition process unit
Electric signal analyzed to obtain the concentration of organic molecule to be detected and be specially：
The main control unit determines organic molecule to be detected using the working sensor curve and the electric signal
Concentration, wherein the working sensor curve is stored in the main control unit.
The present invention provides a kind of plant organic molecule on-line measuring device and method, the present invention is passed using miniature organism
Sensor cell obtains signal, is pre-processed to the signal of acquisition using data acquisition process unit, utilizes main control unit later
Pretreated signal is analyzed to obtain organic molecule concentration to be detected.The insertion of microbiosensor unit is waited for
Be measured in real time in the plant organ of detection, to realize to the online of plant organic molecule, live body, dynamic detection with
And continuous analysis, provide new solution for plant physiology, plant electrophysiology, botany, plant cell biology etc..
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
Obtain other attached drawings according to these attached drawings.
Fig. 1 is the structural schematic diagram of the plant organic molecule on-line measuring device of the preferred embodiment of the present invention；
Fig. 2 is that the plant organic molecule on-line measuring device working signal flow direction of another preferred embodiment of the invention is shown
It is intended to；
Fig. 3 is the plant organic molecule online test method flow chart of another preferred embodiment of the present invention.
Specific implementation mode
Present invention is further described in detail with reference to the accompanying drawings and examples.Following embodiment is for illustrating this hair
It is bright, but cannot be used for limiting the scope of the invention.
A kind of plant organic molecule on-line measuring device, as shown in Figure 1, described device includes microbiosensor list
Member, pumping signal generating unit, data acquisition process unit and main control unit；The main control unit is sent out with the pumping signal
Raw unit connection, the main control unit generate original excitation signal and are transferred to the pumping signal generating unit；The excitation
Signal generating unit is connect with the microbiosensor unit, and the pumping signal generating unit is according to the original excitation
Signal generates pumping signal and passes to the microbiosensor unit；The microbiosensor unit and the number
Connected according to acquisition process unit, the microbiosensor unit under the action of pumping signal with it is to be detected organic
Small molecule reacts, and generates electric signal, and the electric signal of generation is passed to the data acquisition process unit；The data
Acquisition process unit is connect with the main control unit, and the data acquisition process unit passes after being pre-processed to the electric signal
The main control unit is passed, the main control unit carries out the pretreated electric signal that the data acquisition process unit transmits
Analysis obtains the concentration of organic molecule to be detected.
Microbiosensor unit is inserted into plant organ to be detected and is measured in real time, may be implemented to plant
The online of organic molecule, live body, dynamic detection and continuous analysis, be plant physiology, plant electrophysiology, botany,
The new efficient solution of the offers such as plant cell biology.
Further, described device further includes pressure drop temperature compensation unit；Due to device internal resistance in acquiring signal process
Or the pressure drop that brings influences, in the process of running, possible initiation temperature changes, therefore by the pressure drop temperature compensation unit and institute
Microbiosensor unit and main control unit connection are stated, the miniature life is acquired by the pressure drop temperature compensation unit
Pressure drop in the object sensor unit course of work and temperature, and pass to the main control unit.The main control unit is according to institute
The temperature information for stating the transmission of pressure drop temperature compensation unit analyses whether to need to carry out temperature-compensating, swashs if then generating temperature-compensating
It encourages signal and passes to the pumping signal generating unit, as shown in Fig. 2, the pumping signal generating unit is according to the temperature
Compensation incentive signal and the original excitation signal generate the pumping signal.Preferably, the pumping signal generating unit
The temperature-compensating pumping signal and the original excitation signal are superimposed, the pumping signal is generated.The main control unit
Whether the pressure drop information analysis transmitted according to the pressure drop temperature compensation unit needs to carry out voltage compensation, if then exporting correspondence
Offset voltage give the data acquisition process unit, the data acquisition process unit is according to the offset voltage to the electricity
Signal is pre-processed, and by treated, electric signal passes to the main control unit later.
Further, the data acquisition process unit includes that signal amplifies subelement and signal condition subelement, such as
Shown in Fig. 2；The signal amplification subelement is connect with the microbiosensor unit, is used for the electric signal and institute
State the pretreatment that offset voltage is overlapped and amplifies；The signal condition subelement and the signal amplification subelement and
The main control unit connection, for improving amplified electric signal, passes to the main control unit later.
Further, as shown in Fig. 2, the main control unit includes the first digital-to-analogue conversion subunit and the first analog-to-digital conversion
Subelement；The original excitation signal passes to the excitation after carrying out digital-to-analogue conversion by first digital-to-analogue conversion subunit
Signal generating unit；The pretreated electric signal of data acquisition process unit by the first analog-to-digital conversion subelement into
The main control unit is passed to after row analog-to-digital conversion.
The main control unit further includes the second digital-to-analogue conversion subunit, third D/A conversion unit and the second analog-to-digital conversion
Subelement；The temperature-compensating pumping signal is described by being passed to after second digital-to-analogue conversion subunit progress digital-to-analogue conversion
Pumping signal generating unit；The pressure drop of the pressure drop temperature compensation unit acquisition and temperature are by second analog-to-digital conversion
Unit passes to the main control unit after carrying out analog-to-digital conversion；The offset voltage by the third digital-to-analogue conversion subunit into
The microbiosensor unit is passed to after row digital-to-analogue conversion.
Further, the main control unit further includes master control subelement, and sensor work is stored in the master control subelement
Make curve, the concentration of organic molecule to be detected is determined using the working sensor curve and the electric signal.Preferably,
Above-mentioned offset voltage, original excitation signal, temperature-compensating pumping signal are produced after having the master control subelement to handle by analysis
It is raw.The transformed signal of second analog-to-digital conversion subelement and the first transformed signal of analog-to-digital conversion subelement pass to institute
State master control subelement.Preferably above electrical signal is electric current.
Further, above-mentioned first digital-to-analogue conversion subunit, the first analog-to-digital conversion subelement, the second digital-to-analogue conversion are single
Member, third D/A conversion unit and the second analog-to-digital conversion subelement can be a parts for the main control unit, equally also may be used
It is individually present, can be designed according to actual conditions with being detached from the main control unit two.First digital-to-analogue conversion subunit and
Second digital-to-analogue conversion subunit exports the original excitation signal, temperature-compensating pumping signal respectively.
Above-mentioned to have the realization of working sensor curve to determine, drafting is specially：
Configuration concentration gradient be 0,0.01,0.05,0.1,0.5,1.0,5.0,10mM glucose-phosphoric acid buffer (pH=
6.8) solution carries out cyclic voltammetry detection (voltage -1.0~0.6V, sweep speed using the microbiosensor of preparation
50mV/s), the relation curve of one group of concentration and reduction peak current is obtained, microbiosensor working curve, linear side are made
Journey is i=4.8413-98.48501c (current unit nA, concentration unit μM), 0.01~10mM of the range of linearity.
Preferably, the sensor of microbiosensor unit includes reference electrode, working electrode, to electrode.When use
Microbiosensor is inserted into plant roots, stem, leaf or fruit.
Device provided by the invention is integrated with microbiosensor, microelectronics, computer, automatically controls, mathematics, nanometer
Technology etc. provides and directly exists for the scientific research personnel in the fields such as plant physiology, plant electro physiology, botany, Plant Pathology, agronomy
Line detects the tool of small molecule, obtains signal by microbiosensor, small molecule is converted the signal to using main control unit
Concentration, to realize real-time, online, dynamic access little molecular concentration information.
Further, above-mentioned microbiosensor preparation includes the following steps：
1, microbiosensor is prepared using sputtering technology and etching technique in silicon chip substrate, long 4cm, external conduction
The long 5mm in part；
2, microbiosensor current-carrying part is modified into electrode, silver/chlorine by platinum plating using electrochemical deposition method respectively
Change silver-colored reference electrode, plating metal working electrode；
3, gold electrode is polished to smooth with abrasive paper for metallograph, with distilled water and be cleaned by ultrasonic clean；
4, microbiosensor is then placed in progress cyclic voltammetry scan (- 0.1-in 0.5M dilution heat of sulfuric acid
Typical cyclic voltammetric spectrogram 1.6V) is obtained, it is clean to have demonstrate,proved electrode surface；
5,10mM L-cysteine solution is prepared, by its drop coating in gold electrode surfaces, 100% time modification of epidemic disaster in room
20 hours, after clean wash with distilled water, the gold electrode modified；
6, diameter 10nm nano gold sols drop coating is repaiied on 5) the middle gold electrode modified under room epidemic disaster 100%
Decorations 20 hours, distilled water rinses, and obtains gold electrode；
7, enzyme solution drop coating is existed finally by a concentration of 0.1M in the dissolving glucose oxidase containing 70% chitosan solution
6) on gold electrode, 4 DEG C, the modification of humidity 100% time for 24 hours, distilled water is cleaned, and working electrode is obtained, and 4 DEG C of storages are spare.
The method following steps that plant organic molecule is examined online are carried out using above-mentioned apparatus, as shown in Figure 3：
S1, main control unit generate original excitation signal and are transferred to pumping signal generating unit；
S2, the pumping signal generating unit generate pumping signal according to the original excitation signal and pass to miniature life
Object sensor unit；
S3, the microbiosensor unit are sent out under the action of the pumping signal with organic molecule to be detected
Raw reaction, generates electric signal, and the electric signal of generation is passed to data acquisition process unit；
S4, the data acquisition process unit pass to the main control unit, institute after being pre-processed to the electric signal
State the pretreated electric signal that main control unit transmits the data acquisition process unit analyzed to obtain it is to be detected organic
The concentration of small molecule.
Further, the method is further comprising the steps of：
Pressure drop temperature compensation unit acquires pressure drop and temperature during the microbiosensor cell operation, and
Pass to the main control unit；
The main control unit generates temperature-compensating excitation letter according to the temperature information that the pressure drop temperature compensation unit transmits
Number and pass to the pumping signal generating unit, the pumping signal generating unit according to the temperature-compensating pumping signal with
And the original excitation signal generates the pumping signal；
The main control unit exports corresponding offset voltage according to the pressure drop information that the pressure drop temperature compensation unit transmits
To the data acquisition process unit, the data acquisition process unit carries out the electric signal according to the offset voltage
Pretreatment, by treated, electric signal passes to the main control unit later.
Further, in the step S4, pretreatment that the main control unit transmits the data acquisition process unit
The concentration that electric signal afterwards is analyzed to obtain organic molecule to be detected is specially：The main control unit utilizes the sensor
Working curve and the electric signal determine the concentration of organic molecule to be detected, wherein the working sensor curve is stored in
In the main control unit.
Further, further comprising the steps of before being detected using above-mentioned apparatus：Microbiosensor is steamed
Distilled water cleaning after, first respectively detect three parts calibration (0.1,1,5mmol/L) glucose solutions be corrected, finally obtain concentration and
The slope of a curve of current value, when the slope deviation of slope and above-mentioned working curve is being identified as enzymatic activity just within 15%
Often, electrode work is also normal.
The microsensor after correction is inserted into corn stem to be measured when use, cyclic voltammetry scan 10min is carried out, obtains
It is final to obtain corn stem concentration of glucose in conjunction with above-mentioned working curve after taking stable curve.
As a comparison, obtain microsensor detection position tissue samples carry out liquid chromatographic detection, testing result with
The result obtained using the device of the invention or method is compared, and is shown in Table 1.It can be seen that the device of the invention or method detection
Result error is less than 15%, and testing result is reliable, it can be achieved that small molecule on-line checking.
1 test result of table compares
Embodiment of above is merely to illustrate the present invention rather than limitation of the present invention.Although with reference to embodiment to this hair
It is bright to be described in detail, it will be understood by those of ordinary skill in the art that, to technical scheme of the present invention carry out it is various combination,
Modification or equivalent replacement, without departure from the spirit and scope of technical solution of the present invention, the right that should all cover in the present invention is wanted
It asks in range.
Claims (8)
1. a kind of plant organic molecule on-line measuring device, which is characterized in that described device includes microbiosensor list
Member, pumping signal generating unit, data acquisition process unit and main control unit；
The main control unit is connect with the pumping signal generating unit, and the main control unit generates original excitation signal and transmits
To the pumping signal generating unit；
The pumping signal generating unit is connect with the microbiosensor unit, the pumping signal generating unit according to
The original excitation signal generates pumping signal and passes to the microbiosensor unit；
The microbiosensor unit is connect with the data acquisition process unit, and the microbiosensor unit exists
It reacts with organic molecule to be detected under the action of the pumping signal, generates electric signal, and by the electric signal of generation
Pass to the data acquisition process unit；
The data acquisition process unit is connect with the main control unit, the data acquisition process unit to the electric signal into
The main control unit is passed to after row pretreatment, after the pretreatment that the main control unit transmits the data acquisition process unit
Electric signal analyzed to obtain the concentration of organic molecule to be detected；
Described device further includes pressure drop temperature compensation unit；The pressure drop temperature compensation unit and the microbiosensor list
The first and described main control unit connection, the pressure drop temperature compensation unit acquire the microbiosensor cell operation process
In pressure drop and temperature, and pass to the main control unit；
The main control unit generates temperature-compensating pumping signal simultaneously according to the temperature information that the pressure drop temperature compensation unit transmits
The pumping signal generating unit is passed to, the pumping signal generating unit is according to the temperature-compensating pumping signal and institute
It states original excitation signal and generates the pumping signal；
The main control unit exports corresponding offset voltage to institute according to the pressure drop information that the pressure drop temperature compensation unit transmits
Data acquisition process unit is stated, the data acquisition process unit locates the electric signal according to the offset voltage in advance
Reason, by treated, electric signal passes to the main control unit later；
The main control unit includes the first digital-to-analogue conversion subunit and the first analog-to-digital conversion subelement；
The main control unit further includes the second digital-to-analogue conversion subunit, third digital-to-analogue conversion subunit and the second analog-to-digital conversion
Unit；
The temperature-compensating pumping signal is described sharp by being passed to after second digital-to-analogue conversion subunit progress digital-to-analogue conversion
Encourage signal generating unit；
The pressure drop of the pressure drop temperature compensation unit acquisition and temperature carry out modulus by the second analog-to-digital conversion subelement
The main control unit is passed to after conversion；
The offset voltage passes to the miniature organism biography after carrying out digital-to-analogue conversion by the third digital-to-analogue conversion subunit
Sensor cell.
2. the apparatus according to claim 1, which is characterized in that the data acquisition process unit includes that signal amplification is single
Member and signal condition subelement；The signal amplification subelement is connect with the microbiosensor unit, is used for institute
State electric signal and pretreatment that the offset voltage is overlapped and amplifies；The signal condition subelement and the signal
Amplify subelement and main control unit connection, for improving amplified electric signal, passes to the master later
Control unit.
3. the apparatus according to claim 1, which is characterized in that
The original excitation signal passes to the excitation letter after carrying out digital-to-analogue conversion by first digital-to-analogue conversion subunit
Number generating unit；
The pretreated electric signal of data acquisition process unit carries out modulus by the first analog-to-digital conversion subelement and turns
The main control unit is passed to after changing.
4. the apparatus according to claim 1, which is characterized in that the main control unit further includes master control subelement, the master
It is stored with working sensor curve in control subelement, is determined using the working sensor curve and the electric signal to be detected
The concentration of organic molecule.
5. the apparatus according to claim 1, which is characterized in that the electric signal is electric current.
6. a kind of method carrying out plant organic molecule on-line checking using 5 any one of them device described in any one of claim 1 to 5,
It is characterized in that, the described method comprises the following steps：
S1, main control unit generate original excitation signal and are transferred to pumping signal generating unit；
S2, the pumping signal generating unit generate pumping signal according to the original excitation signal and pass to miniature organism and pass
Sensor cell；
S3, the microbiosensor unit occur under the action of the pumping signal with organic molecule to be detected anti-
It answers, generates electric signal, and the electric signal of generation is passed into data acquisition process unit；
S4, the data acquisition process unit pass to the main control unit, the master after being pre-processed to the electric signal
The pretreated electric signal that control unit transmits the data acquisition process unit is analyzed to obtain organic small point to be detected
The concentration of son.
7. according to the method described in claim 6, it is characterized in that, the method is further comprising the steps of：
Pressure drop temperature compensation unit acquires pressure drop and temperature during the microbiosensor cell operation, and transmits
To the main control unit；
The main control unit generates temperature-compensating pumping signal simultaneously according to the temperature information that the pressure drop temperature compensation unit transmits
The pumping signal generating unit is passed to, the pumping signal generating unit is according to the temperature-compensating pumping signal and institute
It states original excitation signal and generates the pumping signal；
The main control unit exports corresponding offset voltage to institute according to the pressure drop information that the pressure drop temperature compensation unit transmits
Data acquisition process unit is stated, the data acquisition process unit locates the electric signal according to the offset voltage in advance
Reason, by treated, electric signal passes to the main control unit later.
8. according to the method described in claim 6, it is characterized in that, in step S4, the main control unit acquires the data
The pretreated electric signal that processing unit transmits is analyzed to obtain the concentration of organic molecule to be detected：
The main control unit determines the concentration of organic molecule to be detected using working sensor curve and the electric signal,
Described in working sensor curve be stored in the main control unit.
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| CN111239225A (en) * | 2020-02-19 | 2020-06-05 | 华南农业大学 | In-situ detection system and method for glucose content in plant fruits |
| CN111693578B (en) * | 2020-06-11 | 2023-01-17 | 中国农业科学院农业信息研究所 | Crop growth information monitoring method and device and manufacturing method thereof |
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