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WO2017018934A1 - Dispositif et procédé pour faire croître un biofilm et mesurer une propriété électrique de celui-ci - Google Patents

Dispositif et procédé pour faire croître un biofilm et mesurer une propriété électrique de celui-ci Download PDF

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Publication number
WO2017018934A1
WO2017018934A1 PCT/SG2016/050345 SG2016050345W WO2017018934A1 WO 2017018934 A1 WO2017018934 A1 WO 2017018934A1 SG 2016050345 W SG2016050345 W SG 2016050345W WO 2017018934 A1 WO2017018934 A1 WO 2017018934A1
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WIPO (PCT)
Prior art keywords
chamber
eis
biofilm
flow cell
electrode
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Ceased
Application number
PCT/SG2016/050345
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English (en)
Inventor
Karinh Emma Josephina EURENIUS
Lars Ake Staffan Kjelleberg
Kai Wei Kelvin LEE
Yong Ling Adelicia LI
Zhiyan Joanne SOH
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National University of Singapore
Nanyang Technological University
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National University of Singapore
Nanyang Technological University
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Publication of WO2017018934A1 publication Critical patent/WO2017018934A1/fr
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/026—Dielectric impedance spectroscopy
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/4833—Physical analysis of biological material of solid biological material, e.g. tissue samples, cell cultures

Definitions

  • the invention relates generally to a device and a method for monitoring the growth of a biofilm and measuring a property of the biofilm, and in particular, to an integrated device wherein the growth and measurement of a property of the biofilm are performed in the same integrated device.
  • an electrochemical impedance spectroscopy EIS is carried out to measure the conduction of the biofilm.
  • a biofilm is an assemblage of surface-associated microbial cells that is enclosed in an extracellular polymeric substance matrix. Biofilms grown on solid surfaces have increasingly gained more attention in recent years because of its wide range of applications, such as industrial, maritime, medical and environment.
  • Electrochemical impedance spectroscopy is a common tool to measure conduction in materials under controlled conditions, i.e. in an atmosphere where the surroundings of the sample, such as gas, temperature or pressure, is known and can be altered and controlled.
  • the immediate problem with the commercially available biofilm growth flow cells is that they are difficult to seal, introducing air bubbles in the growth chambers, causing leakage or contamination.
  • the flow cell itself is primarily suited for microscopy analysis and hence common analytical techniques within materials science (e.g. X-ray Diffraction (XRD), Infrared spectroscopy (IR), Raman spectroscopy, thermal gravimetric analysis (TGA), Scanning Electron Microscopy coupled with Energy Dispersive X-ray spectroscopy (SEM/EDX), Transmission Electron Microscopy (TEM), etc.) cannot be readily used for characterizing the biofilm.
  • XRD X-ray Diffraction
  • IR Infrared spectroscopy
  • Raman spectroscopy Raman spectroscopy
  • TGA thermal gravimetric analysis
  • SEM/EDX Scanning Electron Microscopy coupled with Energy Dispersive X-ray spectroscopy
  • TEM Transmission Electron Microscopy
  • Biofilms are moreover highly sensitive to foreign substances possibly causing contamination and hence the practical set-up of a conventional EIS experiment, introducing amongst others wires and clamps to the biofilm which are not easily sterilized, is not convenient.
  • the EIS measurement needs to be carried out through the biofilm (i.e. across its thickness), and not along the length of the biofilm as is commonly practiced currently, since the biofilm will operate as a membrane or electrolyte in a future electrical device.
  • a device for growing a biofilm and measuring an electrical property of the biofilm may include an electrochemical impedance spectroscopy (EIS) chamber.
  • the device may further include a flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber is entirely housed within the EIS chamber.
  • the flow cell biofilm growth chamber may include an inlet for introducing a flow of nutrient into the flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber may further include an outlet for purging a flow of nutrient out of the flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber may also include a working-electrode-supported substrate located in a flow channel of the flow cell biofilm growth chamber. The working-electrode-supported substrate is adapted for the biofilm growth thereon.
  • the EIS chamber may include an inlet for introducing a flow of inert gas into the EIS chamber.
  • the EIS chamber may further include an outlet for purging a flow of inert gas out of the EIS chamber.
  • the EIS chamber may also include a counter- electrode- supported substrate, wherein the counter-electrode-supported substrate and the working-electrode- supported substrate are adapted for electrical connection to an external impedance measurement apparatus.
  • the EIS chamber may further include one or more openings for insertion of one or more sensors for sensing the environment condition in the EIS chamber.
  • the EIS chamber may further include one or more sensors inserted into the one or more openings.
  • the one or more sensors may include a pH, temperature, pressure, humidity or gas sensor.
  • the flow cell biofilm growth chamber may further include one or more openings for insertion of one or more sensors for sensing the environment condition in the flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber may further include one or more sensors inserted into the one or more openings.
  • the one or more sensors may include a pH, temperature, pressure, humidity or gas sensor.
  • the one or more openings of the EIS chamber may correspond respectively to the one or more openings of the flow cell biofilm growth chamber.
  • the working-electrode-supported substrate may include a material selected from the group consisting of a carbon-supported microscope slide, a silver-supported microscope slide, a gold-supported microscope slide, a fluorinated titanium oxide (FTO) glass, a magnesium, alumina, titanium, copper, silver or gold foil, mesh, tape, plaster, or grid, a paper or plastic printed with carbon, magnesium, titanium, copper, gold, silver or alumina electrode, and a glass microscope cover slip.
  • FTO fluorinated titanium oxide
  • the counter-electrode-supported substrate may include a material selected from a carbon-supported microscope slide, a silver-supported microscope slide, a gold-supported microscope slide, a fluorinated titanium oxide (FTO) glass, a magnesium, alumina, titanium, copper, silver or gold foil, mesh, tape, plaster, or grid, a paper or plastic printed with carbon, magnesium, titanium, copper, gold, silver or alumina electrode, and a glass microscope cover slip.
  • FTO fluorinated titanium oxide
  • a method of growing a biofilm and measuring an electrical property of the biofilm may include providing a device of the first aspect mentioned above.
  • the method may further include introducing a flow of nutrient into the flow cell biofilm growth chamber.
  • the method may also include allowing sufficient time for the growth of a biofilm on the working-electrode-supported substrate located in a flow channel of the flow cell biofilm growth chamber.
  • the method may additionally include electrically connecting the counter-electrode-supported substrate and the working-electrode-supported substrate to an external impedance measurement apparatus to obtain an EIS reading.
  • FIG 1 shows some examples of substrates of various materials onto which biofilms are grown: fluorinated titanium oxide (FTO) glass (top left) can be cut to any desired size and sold in commercially available sheets; various metallic foils, meshes tapes, plasters and grids substrates (top right); printed carbon, silver and alumina electrodes on commercial paper and plastic substrates with biofilms directly grown on the electrode surfaces (mid right); sputtered carbon, silver and gold on microscope slides (bottom right); and microscope cover slips in various sizes (bottom left).
  • FTO fluorinated titanium oxide
  • Figure 2A-2L show photographs of the EIS chamber and the flow cell biofilm growth chamber according to one embodiment.
  • Figure 2A shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the short side of the device.
  • Figure 2B shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the long side of the device.
  • Figure 2C shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the top of the device.
  • Figure 2D shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the short side of the device.
  • Figure 2E shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the long side of the device.
  • Figure 2F shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the top of the device.
  • Figure 2G shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the short side of the device.
  • Figure 2H shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the long side of the device.
  • Figure 21 shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the top of the device.
  • Figure 2J shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the short side of the device.
  • Figure 2K shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the long side of the device.
  • Figure 2L shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the top of the device.
  • Figure 3A-3L show photographs of the present device according to another embodiment.
  • Figure 3A shows a sealed device when viewed from the top of the device.
  • Figure 3B shows a sealed device when viewed from one long side of the device.
  • Figure 3C shows a sealed device when viewed from another long side of the device.
  • Figure 3D shows a sealed device when viewed from one short side of the device.
  • Figure 3E shows a sealed device when viewed from another short side of the device.
  • Figure 3F shows the disassembled device whereby the gas inlet and the gas outlet are removed.
  • Figure 3G shows the disassembled device whereby the working-electrode-supported substrate (including the associated wire clip) and the counter-electrode supported substrate (including the associated wire clip) are removed.
  • Figure 3H shows the disassembled device whereby the sensors and the nutrient inlet and outlet are removed.
  • Figure 31 shows the disassembled device whereby the lid of the EIS chamber is removed.
  • Figure 3J shows the disassembled device whereby the EIS slide adapters are removed.
  • Figure 3K shows the disassembled device whereby the flow cell biofilm growth chamber is removed.
  • Figure 3L shows the disassembled device whereby the lid of the flow cell biofilm growth chamber is removed.
  • Figure 4A-4B show composition and spatial organization of populations for Pseudomonas aeruginosa wildtype PAOl and its over- (+FAP) and non-amyloid expressing (DFAP) mutants, communities of P. protegens Pf5 tagged with CFP (cyan), P. aeruginosa tagged with YFP (yellow) and K. pneumoniae KPl tagged with DsRed (red) triple-, dual- and single species biofilms.
  • DFAP non-amyloid expressing
  • Figure 4A shows a diagram of the flow cell system and chamber (A: growth media, B: peristaltic pump, C: bubble traps, D: flow cell, E: effluent tubes, entering the waste-bottle).
  • the middle section shows a schematic of a flow cell with three channels, tubes connected to both ends of each channel.
  • the micrograph on the right shows a typical confocal laser scanning microscopy (CLSM) image of a biofilm.
  • CLSM confocal laser scanning microscopy
  • Figure 4B shows the biofilm architecture of the P. aeruginosa +FAP strain in medium A shows layers of live and dead (green and red respectively, stained via conventional Syto9 kit) cells (xlOOO magnification).
  • the images provide unique 3D fingerprints of the cell composition.
  • the middle images show the triple species biofilms P. protegens-CFP (blue), P.aeruginosa-YFP (yellow) and K. pneumoniae-OsRed (red) in medium A, representing species composition of the multispecies community.
  • Figure 5A-5C show EIS data collected in the EIS chamber of different embodiments of the present device. Nevertheless, it can be concluded that with the embodiment shown in Figure 2A-2L, valuable and consistent conductivity data (Figure 5A) were collected. This was confirmed with the embodiment shown in Figure 3A-3L at two different time points (24h, ( Figure 5B) and 48h, (Figure 5C)). DESCRIPTION
  • a device for growing a biofilm and measuring an electrical property of the biofilm may include an electrochemical impedance spectroscopy (EIS) chamber.
  • the device may further include a flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber is entirely housed within the EIS chamber.
  • the device of the present invention is an integrated device that enables the growth of a biofilm and the characterization of the thus-grown biofilm via EIS in a single device.
  • the EIS chamber may be covered or sealed (e.g. via screws) with a lid.
  • the flow cell biofilm growth chamber may be covered or sealed (e.g. via screws) with a lid.
  • the flow cell biofilm growth chamber may include an inlet for introducing a flow of nutrient into the flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber may further include an outlet for purging a flow of nutrient out of the flow cell biofilm growth chamber.
  • the flow cell biofilm growth chamber may also include a working-electrode-supported substrate located in a flow channel of the flow cell biofilm growth chamber. The working-electrode-supported substrate is adapted for the biofilm growth thereon.
  • the EIS chamber may include an inlet for introducing a flow of inert gas into the EIS chamber.
  • the EIS chamber may further include an outlet for purging a flow of inert gas out of the EIS chamber.
  • the EIS chamber may also include a counter-electrode-supported substrate, wherein the counter-electrode-supported substrate and the working-electrode-supported substrate are adapted for electrical connection to an external impedance measurement apparatus.
  • EIS slide adapters may be provided inside the EIS chamber.
  • a slid corresponding to the dimension of the cross-section of the counter-electrode-supported substrate, may be provided on one side and an opposing side of the EIS chamber such that the EIS slide adapters and the respective slid on the two opposing sides of the EIS chamber are aligned to allow the insertion of the counter-electrode-supported substrate therethrough.
  • biofilms can be scaled up for large volume production by simply increasing the size of growth vessel. As the bacteria are alive, they will form homogeneous solution which can be integrated to the current battery production line when the electrolyte is applied to battery shell.
  • a biofilm can be directly grown on any substrate located in a flow channel of the flow cell biofilm growth chamber.
  • Commercially available systems for biofilm growth provide non-practical growth channels, besides for microscopic analysis.
  • the device of the present invention allows a support or substrate of various materials to be placed in the growth chamber and taken out after the growth process is completed.
  • existing known growth chambers are sealed shut at the beginning of the growth and it is not possible to open them after the growth is completed, thereby limiting the biofilm analysis to a great extent.
  • FIG 1 shows some examples of substrates of various materials onto which biofilms are grown: fluorinated titanium oxide (FTO) glass (top left) can be cut to any desired size and sold in commercially available sheets; various metallic foils, meshes tapes, plasters and grids substrates (top right); printed carbon, silver and alumina electrodes on commercial paper and plastic substrates with biofilms directly grown on the electrode surfaces (mid right); sputtered carbon, silver and gold on microscope slides (bottom right); and microscope cover slips in various sizes (bottom left).
  • FTO fluorinated titanium oxide
  • Figure 2A-2L show photographs of the EIS chamber and the flow cell biofilm growth chamber according to one embodiment.
  • Figure 2A shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the short side of the device.
  • Figure 2B shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the long side of the device.
  • Figure 2C shows an arrangement (from left to right) of an EIS chamber lid, the EIS chamber, the growth chamber and the growth chamber lid viewed from the top of the device.
  • Figure 2D shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the short side of the device.
  • Figure 2E shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the long side of the device.
  • Figure 2F shows, from left to right, the respective individually assembled EIS chamber and the growth chamber both with their respective lids screwed when viewed from the top of the device.
  • Figure 2G shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the short side of the device.
  • Figure 2H shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the long side of the device.
  • Figure 21 shows, from left to right, the EIS chamber lid, the growth chamber inside the EIS chamber and the growth chamber lid when viewed from the top of the device.
  • Figure 2J shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the short side of the device.
  • Figure 2K shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the long side of the device.
  • Figure 2L shows the assembled growth chamber including its lid inside the EIS chamber including its lid when viewed from the top of the device.
  • Figure 3A-3L show photographs of the present device according to another embodiment, whereby the lid of the EIS chamber is provided with three openings for insertion of three sensors therethrough.
  • sensors are added to the device of the present invention in order to carry out an even more controlled measurement in the growth chamber and the EIS chamber.
  • the sensors are placed close to the inlet with regard to the nutrients, in the middle of the EIS chamber and growth chamber, and close to the outlet of the nutrient flow in order to fully comprehend the internal atmosphere in the device throughout the growth or analysis.
  • a standard diameter of the sensor holes common in today's commercially available pH, temperature and gas (H 2 , 0 2 , N 2 etc.) microsensors is chosen, in order to accommodate any or several environment uptakes to increase flexibility of use.
  • Figure 3A shows a sealed device when viewed from the top of the device.
  • Figure 3B shows a sealed device when viewed from one long side of the device.
  • Figure 3C shows a sealed device when viewed from another long side of the device.
  • Figure 3D shows a sealed device when viewed from one short side of the device.
  • Figure 3E shows a sealed device when viewed from another short side of the device.
  • Figure 3F shows the disassembled device whereby the gas inlet and the gas outlet are removed.
  • Figure 3G shows the disassembled device whereby the working-electrode-supported substrate (including the associated wire clip) and the counter-electrode supported substrate (including the associated wire clip) are removed.
  • Figure 3H shows the disassembled device whereby the sensors and the nutrient inlet and outlet are removed.
  • Figure 31 shows the disassembled device whereby the lid of the EIS chamber is removed.
  • Figure 3J shows the disassembled device whereby the EIS slide adapters are removed.
  • Figure 3K shows the disassembled device whereby the flow cell biofilm growth chamber is removed.
  • FIG. 3L shows the disassembled device whereby the lid of the flow cell biofilm growth chamber is removed.
  • CLSM confocal laser scanning microscopy
  • composition of biofilms on glass was determined with CLSM directly from the flow cell channels.
  • a multi-track mode was used (CFP, YFP and DsRed ⁇ ⁇ : 458, 514 and 561 nm and Xe m : 476, 527 and 584, respectively) to avoid crosstalk between different fluorescent channels.
  • Figure 4A-4B show composition and spatial organization of populations for Pseudomonas aeruginosa wildtype PAOl and its over- (+FAP) and non-amyloid expressing (DFAP) mutants, communities of P. protegens Pf5 tagged with CFP (cyan), P. aeruginosa tagged with YFP (yellow) and K. pneumoniae KPl tagged with DsRed (red) triple-, dual- and single species biofilms.
  • DFAP non-amyloid expressing
  • Figure 4A shows a diagram of the flow cell system and chamber (A: growth media, B: peristaltic pump, C: bubble traps, D: flow cell, E: effluent tubes, entering the waste-bottle).
  • the middle section shows a schematic of a flow cell with three channels, tubes connected to both ends of each channel.
  • the micrograph on the right shows a typical confocal laser scanning microscopy (CLSM) image of a biofilm.
  • CLSM confocal laser scanning microscopy
  • Figure 4B shows the biofilm architecture of the P. aeruginosa +FAP strain in medium A shows layers of live and dead (green and red respectively, stained via conventional Syto9 kit) cells (xlOOO magnification).
  • the images provide unique 3D fingerprints of the cell composition.
  • the middle images show the triple species biofilms P. protegens-CFP (blue), P.aeruginosa-YFP (yellow) and K. pneumoniae -DsRed (red) in medium A, representing species composition of the multispecies community.
  • Figures 4A-4B are results of the CLSM analysis described for the invention in [0065]-[0067].
  • the experimental description on the growth process using the growth chamber of the present device is as follows:
  • P. protegens P. aeruginosa: K. pneumoniae, 5:5: 1 to account for K. pneumoniae 's faster growth rate.
  • Over night cultures were shaken (24 h, 200 rpm, 25°C) and growth was determined via optical density (UV-spectrophotometer) over a 12 h period, to normalize the colony forming units (CFU) prior to inoculation.
  • Cultures were diluted (lxlO 8 cfu ml "1 ), stained with 4', 6-diamidino-2-phenylindole (DAPI) (Yu, W. et al.
  • DAPI 6-diamidino-2-phenylindole
  • Figure 5A-5C show EIS data collected in the EIS chamber of different embodiments of the present device. Nevertheless, it can be concluded that with the embodiment shown in Figure 2A-2L, valuable and consistent conductivity data (Figure 5A) were collected. This was confirmed with the embodiment shown in Figure 3A-3L at two different time points (24h, ( Figure 5B) and 48h, (Figure 5C)).
  • Table 1 shows techniques which are suitable for use in the measurement of a property or analysis of the composition of the biofilm after the growth has completed.
  • FISH Fluorescent in situ Hybridization

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Abstract

L'invention concerne de manière générale un dispositif et un procédé permettant de surveiller la croissance d'un biofilm et de mesurer une propriété du biofilm, et se réfère en particulier à un dispositif intégré dans lequel la croissance et la mesure de la conduction du biofilm sont mises en oeuvre dans le même dispositif intégré. Le dispositif intégré comprend une chambre de croissance de biofilm à cellule d'écoulement entièrement contenue à l'intérieur d'une chambre de spectroscopie d'impédance électrochimique (SIE). La chambre de croissance de biofilm à cellule d'écoulement comprend un substrat supporté sur une électrode de travail, et la chambre de SIE comprend un substrat supporté sur une contre-électrode, les deux substrats étant conçus pour être connectés électriquement à un appareil de mesure d'impédance externe.
PCT/SG2016/050345 2015-07-30 2016-07-21 Dispositif et procédé pour faire croître un biofilm et mesurer une propriété électrique de celui-ci Ceased WO2017018934A1 (fr)

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CN111172024A (zh) * 2019-12-31 2020-05-19 福建农林大学 一种原位表征种间直接电子传递的装置
US20200217778A1 (en) * 2017-05-01 2020-07-09 The Texas A&M University System M-MIC: Microfluidic Microbiologically Influenced Corrosion Model
WO2021092612A1 (fr) * 2019-11-08 2021-05-14 Colgate-Palmolive Company Appareil, système et procédé de test d'échantillons biologiques

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US20200217778A1 (en) * 2017-05-01 2020-07-09 The Texas A&M University System M-MIC: Microfluidic Microbiologically Influenced Corrosion Model
WO2021092612A1 (fr) * 2019-11-08 2021-05-14 Colgate-Palmolive Company Appareil, système et procédé de test d'échantillons biologiques
US12332160B2 (en) 2019-11-08 2025-06-17 Colgate-Palmolive Company Apparatus, system, and method for testing biological samples
CN111172024A (zh) * 2019-12-31 2020-05-19 福建农林大学 一种原位表征种间直接电子传递的装置

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