CN105304919B - A kind of use drop microflow control technique improves the experimental method of the power output of microbiological fuel cell - Google Patents
A kind of use drop microflow control technique improves the experimental method of the power output of microbiological fuel cell Download PDFInfo
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- CN105304919B CN105304919B CN201510388861.6A CN201510388861A CN105304919B CN 105304919 B CN105304919 B CN 105304919B CN 201510388861 A CN201510388861 A CN 201510388861A CN 105304919 B CN105304919 B CN 105304919B
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M8/00—Fuel cells; Manufacture thereof
- H01M8/04—Auxiliary arrangements, e.g. for control of pressure or for circulation of fluids
- H01M8/04082—Arrangements for control of reactant parameters, e.g. pressure or concentration
- H01M8/04186—Arrangements for control of reactant parameters, e.g. pressure or concentration of liquid-charged or electrolyte-charged reactants
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M8/00—Fuel cells; Manufacture thereof
- H01M8/16—Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/50—Fuel cells
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P70/00—Climate change mitigation technologies in the production process for final industrial or consumer products
- Y02P70/50—Manufacturing or production processes characterised by the final manufactured product
Abstract
The present invention proposes a kind of method that use drop microflow control technique improves the power output of microbiological fuel cell, and the method comprising the steps of：（1）Set up drop microfluidic experimental device platform：（2）Set up micro（picoinjection）Experimental solutions injection experimentses device；（3）Set up strain database；（4）Set up host material database；（5）Make with the microbial fuel cell unit for rocking anode；（6）Using step（4）Matrix database in host material and step（3）In strain database in drop strain use step（5）Microbial fuel cell unit, measurement particular substrate material and specified strain under the conditions of, the power output of cell apparatus；Filter out the strain and optimal host material that microbiological fuel cell can be made to reach peak power.The micro-fluidic experimental method of drop that we use has low cost, the advantage that can in high volume test.
Description
Technical field
The present invention relates to the technical field of new energies of sustainable development and regeneration, more particularly to one kind can improve current limit
A kind of experimental method of the low drop microflow control technique of the power output of microbiological fuel cell large-scale promotion application processed.
Background technology
Nearest Nobel Laureate Richard Si Moli (Richard Smalley) often says that " energy problem is the mankind
The ultimate challenge faced ".Scientists are attempted to search out the substitute of oil, such as methane hydrate and coal are changed into
Biogas, but all there is a side effect in these substitutes, they can discharge carbon dioxide, so that aggravate Global Greenhouse Effect,
Although nuclear energy can provide the clean energy resource of non-carbon-emitting, lack at present effective safe method come process cores give up
Gurry.Therefore a series of sustainable alternative energy sources should meet zero carbon emission and reproducible requirement, such as existing at present
Solar energy, wind energy and biomass energy can meet our the following demands to energy.Microbiological fuel cell, is a kind of
Biochemistry can change into the device of electric energy, be the energy of one of them very promising sustainable development.Microbial fuel electricity
Pond is not only a kind of new energy but also can carry out wastewater treatment and carry out biodegradation, with application widely.But
It is it is exported as reproducible clean energy resource and large-scale popularization and application are also faced with many challenges.
Recent researches personnel are improved to microbiological fuel cell in all its bearings can popularization and application, acquirement
Remarkable progress.First, host material is considered as to influence one of important parameter of microbiological fuel cell power output.
It is almost all of to carry out biodegradable organic substance and can serve as the matrix for biological fuel cell to produce electric energy,
Including simple molecule, such as carbohydrate and protein；And human body, it is wastewater produced in animal and food preparation process
The mixture of middle complicated organic matter.Angenent et al. researchs show, include the chemical composition that can change into fuel
Host material is the host material of preferable microbiological fuel cell, and these compositions can also influence thin in anode surface biomembrane
The composition that flora falls, and microbiological fuel cell power output and coulombic efficiency.The host material being typically used for mainly has
Acetate, glucose, lignocellulose biomass, artificial synthesized waste water, Brewage waste water, amylum wastewater, dyestuff gives up
Water, rubbish diffusate, cellulose and chitin, sunshine etc..
Electric charge transporting mechanism is the one of key factor for determining microbiological fuel cell power output.Bacterial cell leads to
Cross respiratory metabolism and produce electric charge, different strains and different electrode potentials can all there are different metabolic processes.Then, these electricity
Lotus is transferred to electrode by physical transfer system (extracellular charge transfer) from cell.This process can be by number of ways come real
Existing, a kind of is that can adhere to the composition of electric charge using soluble charge carrier or certain surface；Another method is by people
Work is added or the medium of bacterial secretory conducts electric charge；Also a kind of is that the nm-class conducting wire that bacterial cell is produced conducts electricity
Lotus.
Also other very many important parameters can influence the power output of microbiological fuel cell, such as anode surface is given birth to
The forming process of thing film can be flowed by electric charge in bacteria culture, environmental factor, anode material and anode potential and biomembrane
Influence.Electrode structure and material are also one of affecting parameters, such as the anode for the multi-pore structure being made of CNT
The power output of microbiological fuel cell can be greatly improved.Nearest Peter et al. is by using adjustment anode working cycle method
To improve power, that is, adjust the time interval that anode is connected with circuit.So overcome the diffusion time due to matrix and electricity
Lotus power reduction caused by the delay of transmission time in extracellular or cell membrane.Although research shows that a variety of microorganisms all may be used
To produce electric current, but the charge density that most of single culture solution are produced is smaller, and the mixed solution meeting of different strain
Improve charge density.Therefore selecting suitable strain or strain combination to have very much the performance for improving microbiological fuel cell must
Will.
The content of the invention
Therefore the present invention improves transmission of the electric charge in electrode surface by new dynamic colloid electrode is designed.We also utilize
Microflow control technique carries out high-speed screening to bacteria culture and host material, and the strain of highest charge density can be produced by selecting
Or strain combination, and host material, improve the performance of microbiological fuel cell eventually through these factors are optimized.
The present invention provides the experiment side that a kind of use drop microflow control technique improves the power output of microbiological fuel cell
Method, comprises the following steps：
(1) drop microfluidic experimental device platform is set up：By aqueous phase solution and oil-phase solution with different in flow rate from different
Direction flows through the drop of the oil-phase solution parcel aqueous phase solution of microchannel generation jointly, and oil-phase solution formation drop shell can be close
Seal cell in any aqueous phase solution, particle or molecule etc.；Culture includes the drop of certain amount strain, and drop is collected micro-
Guan Zhong.
(2) micro (picoinjection) experimental solutions injection experimentses device is set up, acetic acid salt electrolyte is injected into
State in the drop for including bacterial cell, and Dichlorodiphenyl Acetate salting liquid addition organic dyestuff, it is easy to microscopic.
(3) strain database is set up, different bacteria cultures are chosen, each drop includes a kind of strain of identical quantity,
Or the combination of certain several strain, different dye colours are added in bacterial solution drop using the method for step (2) and carried out
Dyeing, so as to be classified, i.e., each color corresponds to a kind of bacterium or bacteria combination, so as to set up strain database.
(4) host material database is set up, each host material is dyed using the method for (2), and each
A kind of host material of color correspondence, so as to set up strain database.
(5) make with the microbial fuel cell unit for rocking anode；Biomembrane is cultivated in specific colloidal droplets, profit
Biomembrane is cultivated with different colloidal droplets, the specific colloidal droplets are to add colloid and the different drop tables of carbon wire
The drop in face；And carbon wire is added inside the colloid of micelle, increase the electric conductivity of micelle, periodically shaken by certain frequency
Dynamic anodic dissolution, to change the biomembrane micelle for being discharged into electric charge of anode surface, and produces the life of electronics in electrolyte
Thing film micelle can be attracted to anode, and the biomembrane micelle being replaced is recovered in the electrolytic solution；The certain frequency is periodically
Rock is by adjusting the frequency and amplitude rocked, adjusting cell recovery time, improve displacement efficiency.
(6) using the host material in the matrix database of step (4) and the drop in the strain database in step (3)
Strain is used under the conditions of the microbial fuel cell unit of step (5), measurement particular substrate material and specified strain, battery dress
The power output put；The biological inoculum of maximum current power can be produced so as to filter out, by bacterial cell, DNA or others
Molecule particles be wrapped in volume from 0.05 be raised to 1 nanoliter of reaction drop slightly in measure power output, so small drop
Volume highly shortened the reaction time so that the measurement of high-speed is to set up different bacterium drop database, so as to filter out
Microbiological fuel cell can be made to reach the strain of peak power；This method biggest advantage is very economical and efficient.
By drop microflow control technique, using the various host materials of screening technique quantitative analysis of step 1~6 in identical
Under the conditions of the current density that can produce, so as to pick out optimal host material, improve the output work of microbiological fuel cell
Rate.Various host materials are wrapped in the reaction drop of very small volume according to different concentration and composition, host material is set up
Database.It can realize daily to 108Individual reaction drop is made and measured.By this method, can with it is minimum into
This carries out experiment measurement analysis to uncommon somewhat expensive host material.
The prioritization scheme obtained by above experimental data, is tested in microbial fuel cell unit, through excessive
The different strain of amount, the combination of matrix and drop biomembrane obtains optimal design, makes the output of microbiological fuel cell
Power reaches maximum.
Further, the aqueous phase solution in the step (1) uses Luria-Bertani Bacteria Culture solution, every 1 liter
Luria-Bertani solution includes 10 grams of peptones, 5 grams of sodium chloride, 5 grams of yeast extracts and 1 liter of distilled water, and mixture is molten
Liquid carries out high-temperature sterilization, can be stored into standby in subzero 4 degree of refrigerator.
Further, bacteria culture different in the step (3) includes ground bacillus, and Shewanella, Pseudomonas aeruginosa produces amber
Amber acid Actinobacillus, Aeromonas hydrophila, Bacillus foecalis alkaligenes, clostridium butyricum, desulfovibrio desulfurican, Erwinia, Escherichia coli,
Gluconobacter oxydans, pneumobacillus, Lactobacillus plantarum, proteus mirabilis, thermophilic sweet microorganism, streptococcus lactis.Each liquid
A kind of strain comprising identical quantity, measure they produce electric charges number, so as to classify to strain drop, similar approach
The drop of different strain combination is tested, the charge number of generation is classified.Different dyestuffs are added in bacterial solution drop
Color is dyed.Data analysis is carried out, the database that different strain produces electric charge is obtained.
Further, the organic dyestuff of the step (2) be bipseudoindoxyl dye (a kind of organic dyestuff extracted from plant,
Chemical expression is C16H10N2O2), it would however also be possible to employ other color dyes,
Further, the step (4), host material is acetate, arabitol, glucose, carboxymethyl cellulose, fiber
Crude granule, cornstalk biological matter, cysteine, 1,2- dichloroethanes, alcohol or furfural.And compare and produced under various concentrations
Charge number, sets up electrolyte host material database.
Further, colloidal droplets different in the step (5) are single face colloidal droplets, double layer surface colloidal droplets
With one kind in three layers of colloid surface drop,
Further, shaking amplitude and slosh frequency are needed according to corresponding output power measurement value in the step (5)
It is determined that, it is ensured that output power measurement value be in the range of output power measurement value above and below peak in 5~20% value range (preferably
Ensure output power measurement value in above and below peak in the range of output power measurement value 10% value range), the output work
Rate measured value be microbial fuel cell unit, measurement particular substrate material and specified strain under the conditions of, cell apparatus it is defeated
Go out power.
Further, cell recovery time is 3~10s in the step (5)；It is preferred that, cell recovery time can shorten
To 3~5s.
Such as periodic reverse each second is rocked once, it is ensured that output power measurement value is in the range of output power measurement value
Above and below peak in 10% value range, to change the biomembrane micelle for being discharged into electric charge of anode surface, and electrolyte
The middle biomembrane micelle for producing electronics can be attracted to anode, and the biomembrane micelle being replaced is recovered in the electrolytic solution.We
Referred to as dynamic colloid electrode, is that bacterial biof iotalm grows in drop, cell can undergo three kinds of bacteriums as shown in schematic diagram 4
The conversion of type, contact of the cell with anode is controlled by cell growth in drop, conveniently displaces anode surface release
Cell after complete electric charge is recovered into electrolyte, and the cell liquid of the generation electric charge of tonic newly drips to anode surface.Using this
The cell recovery time that the method for invention is obtained is 5s or so, will be loaded well below in current microbiological fuel cell in document
15 seconds recovery times in cycle, so as to increase the time that anode electric charge is maintained at peak state.Overcome because electric charge is in electricity
Power reduction caused by the transmission recovery time of pole, can be greatly improved the power output of microbial fuel cell unit.
The present invention will improve the power output of microbiological fuel cell using drop microflow control technique, mainly including following
Three aspects：
A) design shadows of the duty cycle to microbiological fuel cell performance in recent years is optimized to microbiological fuel cell
Sound is of concern.Optimize power output with the time disconnected by adjusting external circuit and being connected with anode.Its principle be because
The transmission in microbiological fuel cell of matrix and electric charge and their transmission in biomembrane need diffusion time.We
Project will improve this problem using microflow control technique.We replace traditional glue with the biomembrane being grown in colloidal droplets
The biomembrane of anode surface is attached to, and wire such as CNT is added in the colloidal droplets that growth has biomembrane, to increase
Plus the electric conductivity of biomembrane micelle.Each colloidal droplets with biomembrane so with electrode contact have identical structure,
And because bacterial cell is wrapped up by colloid, therefore each biomembrane micelle is separate.We are molten by rocking anode
Liquid, to change the biomembrane micelle of anode surface, the biomembrane micelle being replaced is recovered in the electrolytic solution, we term it
Dynamic colloid electrode.The cell recovery time being obtained by the present invention is adopted at 3~10 seconds, it is micro- at present well below in document
15 seconds recovery times in biological fuel cell in the duty cycle, thus increase anode electric charge be maintained at peak state when
Between.Overcome because electric charge is in power reduction caused by electrode transmission recovery time；Microbial fuel cell unit can be greatly improved
Power output.
B) optimizing microorganism fungus kind or bacterium colony using drop microflow control technique, we are adopted using drop microflow control technique
The biological inoculum or strain combination of high current density can be produced by being filtered out with superelevation speed.Nearest research shows, although
Very many bacteriums can make microbiological fuel cell produce electric current, but most of be obtained with the nutrient solution of single culture
Current density ratio is relatively low.To improve current density, it would be desirable to which high current density can be produced by quantifying any strain.This hair
Bright use drop microflow control technique sets up the measuring table of ultra-high throughput, and the biological bacteria of maximum current power can be produced by filtering out
Kind, this method biggest advantage is very economical and efficient.We can be by bacterial cell, DNA or other molecules
Grain be wrapped in volume from 0.05 be raised to 1 nanoliter of reaction drop slightly in measure, so small droplet size is greatly
Shorten the reaction time so that the measurement of high-speed is possibly realized.So we can just set up different bacterium drop database,
So as to filter out the strain that microbiological fuel cell can be made to reach peak power.
C) the superelevation speed of host material screens in microbiological fuel cell, and host material is considered as one of them
The factor of very important influence power output.The species of host material widely, can be single component material or
It is the combination of various organic matters in waste water.Up to the present, researcher, which has been set out, can be used in microbiological fuel cell hair
The inventory of the host material of electricity, but be due to different experimental situations, experiment condition, electrode material and surface characteristic and difference
Strain select, it is difficult to influence of these host materials of quantitative comparison to power output.We will pass through drop microflow control technique
To solve this problem, it can be produced at identical conditions using the various host materials of ultrahigh speed screening technique quantitative analysis
Current density, so as to pick out optimal host material.We can be various host materials according to different concentration and composition
It is wrapped in the reaction drop of very small volume, sets up host material database, it is possible to achieve 108 reaction drops is carried out daily
Make and measure.By this method, we can be carried out real with minimum cost to uncommon somewhat expensive host material
Test amount is analyzed.
Brief description of the drawings
Traditional microbiological fuel cell experimental provision that Fig. 1 is prepared for the present invention；
Fig. 2 a have the microbial fuel cell unit for rocking anode for what the present invention was designed；
The drop micelle biomembrane having in the microbial fuel cell unit for rocking anode that Fig. 2 b design for the present invention
Enlarged drawing；
Fig. 3 A are the experimental provisions for making microlayer model；
Fig. 3 B prepare the experimental provision that growth has the drop of biomembrane；
Fig. 3 C are by the drop microscope photograph of Fig. 3 B double emulsifieds produced；
Fig. 4 A are the cell types of the different growth phase of biomembrane in drop；
Fig. 4 B are the microscope photographing pictures of cell type of the biomembrane at 2 hours in drop；
Fig. 4 C are the microscope photographing pictures of cell type of the biomembrane at 24 hours in drop；
Fig. 4 D are the microscope photographing pictures of cell type of the biomembrane at 72 hours in drop；
The drop containing bacterial biof iotalm shown in Fig. 5 A with double emulsion layers is contrasted than other drops without biomembrane
Microscope photograph；
The drop containing bacterial biof iotalm shown in Fig. 5 B with single emulsion layer is contrasted than other drops without biomembrane
Microscope photograph；
Different double influences of the thickness to biofilm formation for being emulsified into drop shell shown in Fig. 5 C, picture is microscope photographing
Picture of the drop of three kinds of drop thickness of the shells 24 hour moment；
Fig. 6 is used for carrying out bacterium for the present invention and host material carries out the micro fluidic device of ultrahigh speed screening；
The experimental provision for preparing drop shown in Fig. 6 a for the general's choosing of superelevation speed；
1,000,000 a diameter of 25 microns of drops are stored in micro-pipe shown in Fig. 6 b；
Drop shown in Fig. 6 c sprays into experimental provision again；
Micro- microscale droplets sprays into experimental provision again shown in Fig. 6 d；
Entrainment trap shown in Fig. 6 e；
The screening plant of drop shown in Fig. 6 f；
Using the foundation in drop storehouse during microfluidic methods progress ultrahigh speed screening shown in Fig. 7 a；
It is the screening that drop storehouse is used to detect the micro constitutent of antiseptic shown in Fig. 7 b.
Embodiment
Embodiments of the invention are described below in detail, the example of the embodiment is shown in the drawings, wherein from beginning to end
Same or similar label represents same or similar element or the element with same or like function.Below with reference to attached
The embodiment of figure description is exemplary, is only used for explaining the present invention, and is not considered as limiting the invention.
A kind of use drop microflow control technique improves the experimental method of the power output of microbiological fuel cell, including following
Step：
(1) drop microfluidic experimental device platform is set up：By aqueous phase solution and oil-phase solution with different in flow rate from different
Direction flows through the drop of the oil-phase solution parcel aqueous phase solution of microchannel generation jointly, and oil-phase solution formation drop shell can be close
Seal cell in any aqueous phase solution, particle or molecule etc.；Culture includes the drop of certain amount strain, and drop is collected micro-
Guan Zhong.Aqueous phase solution therein uses Luria-Bertani Bacteria Culture solution, and every 1 liter of Luria-Bertani solution includes 10
Mixture solution, is carried out high-temperature sterilization, can stored by gram peptone, 5 grams of sodium chloride, 5 grams of yeast extracts and 1 liter of distilled water
It is standby into subzero 4 degree of refrigerators.
(2) micro (picoinjection) experimental solutions injection experimentses device is set up, acetic acid salt electrolyte is injected into
State in the drop for including bacterial cell, and Dichlorodiphenyl Acetate salting liquid addition organic dyestuff, it is easy to microscopic.It is therein organic
Dyestuff is that (a kind of organic dyestuff extracted from plant, chemical expression is C to bipseudoindoxyl dye16H10N2O2), it would however also be possible to employ its
His color dye,
(3) strain database is set up, different bacteria cultures are chosen, each drop includes a kind of strain of identical quantity,
Or the combination of certain several strain, different dye colours are added in bacterial solution drop using the method for step (2) and carried out
Dyeing, so as to be classified, i.e., each color corresponds to a kind of bacterium or bacteria combination, so as to set up strain database.Institute
Stating bacteria cultures different in step (3) includes ground bacillus, Shewanella, Pseudomonas aeruginosa, Actinobacillus succinogenes, thermophilic aqueous vapor
Monad, Bacillus foecalis alkaligenes, clostridium butyricum, desulfovibrio desulfurican, Erwinia, Escherichia coli, gluconobacter oxydans, pneumonia bar
Bacterium, Lactobacillus plantarum, proteus mirabilis, thermophilic sweet microorganism, streptococcus lactis.Each liquid includes a kind of bacterium of identical quantity
Kind, measure they produce electric charges number, so as to classify to strain drop, the liquid that similar approach test different strain is combined
Drop, the charge number of generation is classified.Different dye colours are added in bacterial solution drop to be dyed.Carry out data
Analysis, obtains the database that different strain produces electric charge.
(4) host material database is set up, each host material is dyed using the method for (2), and each
A kind of host material of color correspondence, so as to set up strain database.Host material is acetate, arabitol, glucose, carboxymethyl
Cellulose, cellulose grain, cornstalk biological matter, cysteine, 1,2- dichloroethanes, alcohol or furfural.And ratio is less
With charge number is produced under concentration, electrolyte host material database is set up.
(5) make with the microbial fuel cell unit for rocking anode；Biomembrane is cultivated in specific colloidal droplets, profit
Biomembrane is cultivated with different colloidal droplets, the specific colloidal droplets are to add colloid and the different drop tables of carbon wire
The drop in face；And carbon wire is added inside the colloid of micelle, increase the electric conductivity of micelle, periodically shaken by certain frequency
Dynamic anodic dissolution, to change the biomembrane micelle for being discharged into electric charge of anode surface, and produces the life of electronics in electrolyte
Thing film micelle can be attracted to anode, and the biomembrane micelle being replaced is recovered in the electrolytic solution；The certain frequency is periodically
Rock is by adjusting the frequency and mode rocked, adjusting cell recovery time, improve displacement efficiency.Shaking amplitude and rock frequency
Rate need according to corresponding output power measurement value determine, it is ensured that output power measurement value be output power measurement value in the range of most
(preferably ensure that output power measurement value is peak in the range of output power measurement value in 5~20% value range above and below high level
Up and down in 10% value range), the output power measurement value be microbial fuel cell unit, measurement particular substrate material and
Under the conditions of specified strain, the power output of cell apparatus.By the selection of specific host material and specified strain, and it is above-mentioned
It is that 3~10s wherein different colloidal droplets are single face colloidal droplets, double layer surface colloid that method, which can adjust cell recovery time,
One kind in drop and three layers of colloid surface drop.
Such as periodic reverse each second is rocked once, it is ensured that output power measurement value is in the range of output power measurement value
Above and below peak in 10% value range, to change the biomembrane micelle for being discharged into electric charge of anode surface, and electrolyte
The middle biomembrane micelle for producing electronics can be attracted to anode, and the biomembrane micelle being replaced is recovered in the electrolytic solution.We
Referred to as dynamic colloid electrode, as shown in schematic diagram 4；It is that bacterial biof iotalm grows in drop, cell can undergo three kinds of bacteriums
The conversion of type, contact of the cell with anode is controlled by cell growth in drop, conveniently displaces anode surface release
Cell after complete electric charge is recovered into electrolyte, and the cell liquid of the generation electric charge of tonic newly drips to anode surface.
It is 5s or so to adopt the cell recovery time being obtained by the present invention, will be well below current microorganism in document
15 seconds recovery times in fuel cell in the duty cycle, so as to increase the time that anode electric charge is maintained at peak state.Gram
The defeated of microbial fuel cell unit can be greatly improved because electric charge is in power reduction caused by electrode transmission recovery time in clothes
Go out power.
(6) using the host material in the matrix database of step (4) and the drop in the strain database in step (3)
Strain is used under the conditions of the microbial fuel cell unit of step (5), measurement particular substrate material and specified strain, battery dress
The power output put；The biological inoculum of maximum current power can be produced so as to filter out, by bacterial cell, DNA or others
Molecule particles be wrapped in volume from 0.05 be raised to 1 nanoliter of reaction drop slightly in measure power output, so small drop
Volume highly shortened the reaction time so that the measurement of high-speed is to set up different bacterium drop database, so as to filter out
Microbiological fuel cell can be made to reach the strain of peak power；This method biggest advantage is very economical and efficient.
By drop microflow control technique, existed using the various host materials of ultrahigh speed screening technique quantitative analysis of step 1~6
The current density that can be produced under the same conditions, so as to pick out optimal host material.Various host materials according to not
Same concentration and composition is wrapped in the reaction drop of very small volume, sets up host material database.It can realize daily to 108
Individual reaction drop is made and measured.By this method, can be with minimum cost to uncommon somewhat expensive matrix
Material carries out experiment measurement analysis.
The prioritization scheme obtained by above experimental data, is tested in microbial fuel cell unit, through excessive
The different strain of amount, the combination of matrix and drop biomembrane obtains optimal design, makes the output of microbiological fuel cell
Power reaches maximum.
The main equipment preparation method and step for describing to be tested according to the present invention below in conjunction with accompanying drawing.
It is the laboratory installation of microbiological fuel cell as shown in Figure 1.
The part of Fig. 1 microbiological fuel cells mainly includes anode, negative electrode and electrolyte, is attached to the micro- of anode tap
Host material in bio-catalytical oxidation anode chamber, microbial cell discharges electronics by breathing to anode, these electronics by
External circuit is delivered to negative electrode to produce electric current.Microbial cell often produces an electronics will be while produce a proton, matter
Son is delivered to negative electrode to maintain electric current by electrolyte.Under normal circumstances, electronics and proton negative electrode and oxygen platinum catalysis
Reacted under effect generation water anodes and negative electrode reaction it is as follows, here from being used as host material exemplified by acetate.
Anodic reaction：CH3COO-
Cathodic reaction：O2+4e-+4H+→2H2O (2)
Fig. 2 is that having of designing of the present invention rocks the microbial fuel cell unit of anode,
Fig. 2 a. are microbiological fuel cell schematic diagrames, and left side is anode reaction room, and right side is cathode reaction chambers, it is middle by
PEM is isolated.Electrolyte in anode reaction room is in vibrating state, the biology that anode surface is wrapped up by bacterium colloid
Film liquid drop is covered.
Fig. 2 b. are the enlarged diagrams of biomembrane micelle, and biofilm development uniformly divides in the inner surface of micelle in colloid
Cloth wire (carbon black), increases the electric conductivity of biomembrane micelle.We are by rocking the electrolyte in anode chamber, to replace sun
The biomembrane micelle on pole surface, the biomembrane micelle for being set to change is recovered in the electrolytic solution.
It is to prepare the device that biomembrane is grown in micelle shown in Fig. 3.
Fig. 3 A are the experimental provisions for making microlayer model, and W represents aqueous phase, and O represents oil phase, and we are micro-fluidic using glass tube
Experimental provision, when aqueous phase and oil phase with certain speed flow through specific glass tube fixture, will form oil phase and be wrapped in aqueous phase
Microlayer model, the size of drop determines according to the radius of glass tube.
Fig. 3 B are replaced the aqueous phase in interior conduit in Fig. 3 A by the solution and bacillus subtilis cell of culture biomembrane, wherein carefully
The density of born of the same parents is OD600Outside=1, the solution of culture biomembrane is MSgg solution, and composition is 5mM potassium phosphates (pH 7)/100mM 3-
(N-morpholino) propane sulfonate (MOPS) (pH 7)/2mM magnesium chloride/700 μM CaCl2/50μM MnCl2/
50μM FeCl2/1μM ZnCl2/ 2 μM of thiamine/0.5%glycerol/0.5%glutamate/50g/ml
Tryptophan/50g/ml phenylalanine and water, above-mentioned chemical composition need to sterilize or high-temperature sterilization.In pipeline
Oil phase be liquid phase in silicone oil and surfactant ((w/v) DC749 of 10cSt DC200 PDMS with 2%), Outer Tube
Composition be MSgg solution wherein comprising 10% (w/v) polyvinyl alcohol (polyvinylalcohol, 87%-89%hydrolyzed,
Mw13000-23000,Sigma-Aldrich).The glass tube for the taper that two external diameters are 1 millimeter, which is placed on an internal diameter, is
Drop is made in 1.05 millimeters of rectangular tube.
Fig. 3 C are by the drop microscope photograph of Fig. 3 B double emulsifieds produced.Scale in figure is 200 microns.
It is the microscope photograph for preparing growth biomembrane in drop by Fig. 3 shown in Fig. 4.We use double labelling in experiment
(the amyE of Bacillus subtilis strain 3610::PtapA-yfp,lacA::Phag-cfp)
Fig. 4 A are the cell types of the different growth phase of biomembrane in drop.In several hours initial bacterial cells
It is the motility cell of free state, as shown in Fig. 3 A left hand views；Cell differentiation is into secretion matrix 12-24 hours when
Cell, as shown in the figure in the middle of Fig. 3 A；It was changed into spore cell at 48-72 hours, as shown in Fig. 3 A right part of flg.
Fig. 4 B are the microscope photographing pictures of the cell type of the different growth phase of biomembrane in drop.
As shown in Fig. 4 B Image to lefts at 2 hours, blueness (P is presented in most cellshag- cfp, the whip of hag generations here
Hair), it is shown to be motility cell.The picture of microscope photographing is the central cross-section of drop.
As shown in Fig. 4 B intermediate pictures at 24 hours, yellow is presented in most cells, and it is secretion base to show cells switch
The cell of matter, and in the inner surface formation biomembrane of drop.The picture of microscope photographing is close to the bottom of drop.
As shown in Fig. 4 B Image to rights at 72 hours, most cells do not have fluorescence display, and in drop bottom
Form the spore cell of soccer star.The scale of picture is 50 microns.Cell and anode are controlled in drop by cell growth
Contact, conveniently displaces anode surface and releases the cell after electric charge and recovered into electrolyte, the generation electric charge of tonic newly
Cell liquid drip to anode surface.
Influence of the different drop thickness of the shells to biomembrane is represented shown in Fig. 5.
There is the drop containing bacterial biof iotalm of double emulsion layers than other drops without biomembrane, volume shown in Fig. 5 A
It can reduce, show that the moisture in the also drop of biomembrane flows out from the oil reservoir shell of drop.
There is the drop containing bacterial biof iotalm of single emulsion layer than other drops without biomembrane, volume shown in Fig. 5 B
Also it can reduce, show equally there is moisture to be flowed out from the oil reservoir shell of drop.
Different double influences of the thickness to biofilm formation for being emulsified into drop shell shown in Fig. 5 C, picture is microscope photographing
The drop of three kinds of drop thickness of the shells is in the picture at 24 hour moment, and biomembrane can all be had by showing the drop of three kinds of drop thickness of the shells.
Scale in wherein 4A represents 200 microns, and the scale in 4B represents 25 microns, and the scale in 4A represents 50 microns.
Using drop preparation facilities during microfluidic methods progress ultrahigh speed screening shown in Fig. 6.
The experimental provision of drop is prepared shown in Fig. 6 a.A kind of liquid solution (W) flows through microflow channels jointly with oil-phase solution (O)
The drop (WO) of road formation oil phase internal package aqueous phase.Aqueous phase in drop can include cell, particle and molecule.
1,000,000 a diameter of 25 microns of drop storages (about 20 microlitres of shared volume) have 200 microlitres shown in Fig. 6 b
In micro-pipe.
Drop shown in Fig. 6 c sprays into experimental provision again, takes out the drop (WO) prepared by Fig. 6 a shown devices, then spray into micro-
Flow duct facility.In exit, we add oil phase (O) to separate compact arranged drop.
Micro- microscale droplets shown in Fig. 6 d spray into experimental provision again, and the drop (WO) prepared by Fig. 6 a shown devices is from a left side
Side is by T-shaped microtube device, during blue liquid (W) injects drop above T-shaped microtube device, under T-shaped microtube device
The heavy black of side represents electrode, for making drop shell unstability, so as to be blue liquid injection drop.It is thus from T-shaped micro-pipe
The new drop produced on the right side of duct device includes blue liquid, and blue liquid can be rapidly diffused into whole drop.
Entrainment trap shown in Fig. 6 e.The drop (WO) prepared by Fig. 5 a shown devices passes through T-shaped microtube device, liquid
Drop can resolve into two drops, can pass through the big of the new drop for the geometric parameter control decomposition for adjusting T-shaped microtube device
It is small.
The screening plant of drop shown in Fig. 6 f, the black drop of the device leftmost side includes blue dyes, white drop
Include the particle of fluorescence labeling, two kinds of droplets mixings are together.White circle below figure Green arrow is laser facula,
For being screened to drop.Heavy black below device represents electrode and is used for exciting fluorescence display, for detecting light color
Drop containing fluorescence labeling.Scale in figure represents 100 microns.Arrow represents the flow direction of different phase liquid.
Using the foundation and the screening of minor constituent in drop storehouse during microfluidic methods progress ultrahigh speed screening shown in Fig. 7.
Shown in Fig. 7 a using Fig. 6 a drop preparation facilities and include it is multiple can place the plate of lot of trace liquid well,
We can be fabricated to drop to the liquid in each well of plate simultaneously, and the various drops comprising different solutions are then stored in one
Rise in the well for forming each storing solution of drop storehouse and contain and replaced in specific bar code, Fig. 7 a with different colors, shape
Into corresponding each drop in also there is bar code corresponding with solution in well, so as to for screening.
It is the screening that drop storehouse is used to detect the micro constitutent of antiseptic shown in Fig. 7 b.In the drop storehouse prepared by Fig. 7 a
Drop drop prepare in sprayed into again, and give in each drop using Fig. 6 d method and inject single bacterial cell, carry out
Culture, then by detecting that the life and death of the cell in drop is screened to drop, the solution for finding out specific bar code resists
Bacterium property.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means to combine specific features, structure, material or the spy that the embodiment or example are described
Point is contained at least one embodiment of the present invention or example.In this manual, to the schematic representation of above-mentioned term not
Necessarily refer to identical embodiment or example.Moreover, specific features, structure, material or the feature of description can be any
One or more embodiments or example in combine in an appropriate manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that：Not
In the case of departing from the principle and objective of the present invention a variety of change, modification, replacement and modification can be carried out to these embodiments, this
The scope of invention is by claim and its equivalent limits.
Claims (9)
1. a kind of method that use drop microflow control technique improves the power output of microbiological fuel cell, comprises the following steps：
(1) drop microfluidic experimental device platform is set up：By aqueous phase solution and oil-phase solution with it is different in flow rate from different directions
The oil-phase solution for flowing through microchannel generation jointly wraps up the drop of aqueous phase solution, and oil-phase solution formation drop shell can be sealed
Cell in any aqueous phase solution, particle or molecule；Culture includes the drop of certain amount strain, and drop is collected in micro-pipe
In；
(2) micro picoinjection experimental solutions injection experimentses device is set up, acetic acid salt electrolyte is injected into above-mentioned include
Have in the drop of bacterial cell, and Dichlorodiphenyl Acetate salting liquid addition organic dyestuff, it is easy to microscopic；
(3) strain database is set up, different bacteria cultures are chosen, each drop includes a kind of strain of identical quantity, or
The combination of certain several strain, adds different dye colours using the method for step (2) in bacterial solution drop and is dyed,
So as to be classified, i.e., each color corresponds to a kind of bacterium or bacteria combination, so as to set up strain database；
(4) host material database is set up, each host material is dyed using the method for (2), and each color
A kind of host material of correspondence, so as to set up host material database；
(5) make with the microbial fuel cell unit for rocking anode；Biomembrane is cultivated in specific colloidal droplets, using not
With colloidal droplets cultivate biomembrane, the specific colloidal droplets are the different droplet surfaces for adding colloid and carbon wire
Drop；And carbon wire is added inside the colloid of micelle, increase the electric conductivity of micelle, sun is periodically rocked by certain frequency
Pole solution, to change the biomembrane micelle for being discharged into electric charge of anode surface, and produces the biomembrane of electronics in electrolyte
Micelle can be attracted to anode, and the biomembrane micelle being replaced is recovered in the electrolytic solution；The certain frequency is periodically rocked
It is by adjusting the frequency and amplitude rocked, adjusting cell recovery time, improve displacement efficiency；
(6) using the host material in the matrix database of step (4) and the drop strain in the strain database in step (3)
Using the microbial fuel cell unit of step (5), under the conditions of measurement particular substrate material and specified strain, cell apparatus
Power output；The biological inoculum of maximum current power can be produced so as to filter out, by bacterial cell, DNA or other molecules
Particle be wrapped in volume from 0.05 be raised to 1 nanoliter of reaction drop slightly in measure power output, so small droplet size
It highly shortened the reaction time so that the measurement of high-speed, can so as to filter out to set up different bacterium drop database
Microbiological fuel cell is set to reach the strain of peak power；Various host materials are wrapped in very according to different concentration and composition
In the reaction drop of small size, at identical conditions can using the various host materials of screening technique quantitative analysis of above-mentioned steps
The current density enough produced, picks out optimal host material；
Aqueous phase solution in the step (1) uses Luria-Bertani Bacteria Culture solution, and every 1 liter of Luria-Bertani is molten
Liquid includes 10 grams of peptones, 5 grams of sodium chloride, 5 grams of yeast extracts and 1 liter of distilled water, and mixture solution is carried out high-temperature sterilization,
It is stored into standby in subzero 4 degree of refrigerator.
2. use drop microflow control technique according to claim 1 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：Different bacteria cultures includes ground bacillus in the step (3), and Shewanella, Pseudomonas aeruginosa produces amber
Sour Actinobacillus, Aeromonas hydrophila, Bacillus foecalis alkaligenes, clostridium butyricum, desulfovibrio desulfurican, Erwinia, Escherichia coli, oxygen
Change gluconobacter sp, pneumobacillus, Lactobacillus plantarum, proteus mirabilis, thermophilic sweet microorganism, streptococcus lactis.
3. use drop microflow control technique according to claim 1 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：The organic dyestuff of the step (2) is bipseudoindoxyl dye or other color dyes.
4. use drop microflow control technique according to claim 1 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：The step (4), host material is acetate, arabitol, glucose, carboxymethyl cellulose, cellulose
Particle, cornstalk biological matter, cysteine, 1,2- dichloroethanes, alcohol or furfural.
5. use drop microflow control technique according to claim 1 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：Different colloidal droplets are single face colloidal droplets in the step (5), double layer surface colloidal droplets and
One kind in three layers of colloid surface drop.
6. use drop microflow control technique according to claim 1 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：Shaking amplitude and slosh frequency need true according to corresponding output power measurement value in the step (5)
It is fixed, it is ensured that output power measurement value is in the range of output power measurement value above and below peak in 5~20% value range, described defeated
Go out power measurement values for microbial fuel cell unit, under the conditions of measurement particular substrate material and specified strain, cell apparatus
Power output.
7. use drop microflow control technique according to claim 6 improves the side of the power output of microbiological fuel cell
Method, it is characterised in that：Shaking amplitude and slosh frequency need true according to corresponding output power measurement value in the step (5)
It is fixed, it is ensured that output power measurement value is in above and below peak in the range of output power measurement value 10% value range.
8. the use drop microflow control technique according to claim 1 or 6 improves the power output of microbiological fuel cell
Method, it is characterised in that：Cell recovery time is 3~10s in the step (5).
9. the use drop microflow control technique according to claim 1 or 6 improves the power output of microbiological fuel cell
Method, it is characterised in that：Cell recovery time is 3~5s in the step (5).
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| US4652501A (en) * | 1984-07-24 | 1987-03-24 | King's College London | Operation of microbial fuel cells |
| CN101789515A (en) * | 2010-03-17 | 2010-07-28 | 哈尔滨工程大学 | Method for improving electron transfer capacity and output power of microbial fuel cell |
| CN104078692A (en) * | 2013-11-27 | 2014-10-01 | 中科宇图天下科技有限公司 | Control device and control method for microbial fuel cell |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4652501A (en) * | 1984-07-24 | 1987-03-24 | King's College London | Operation of microbial fuel cells |
| CN101789515A (en) * | 2010-03-17 | 2010-07-28 | 哈尔滨工程大学 | Method for improving electron transfer capacity and output power of microbial fuel cell |
| CN104078692A (en) * | 2013-11-27 | 2014-10-01 | 中科宇图天下科技有限公司 | Control device and control method for microbial fuel cell |
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