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<table><tbody><tr><td>Adapter fitting (Elveflow, 6-40 to 1/4-28)</td><td>Darwin Microfluidics</td><td>LVF-KFI-08</td><td>Used to interface between flow sensor and flangeless fittings.</td></tr><tr><td>Agar (Fisher BioReagents)</td><td>Fisher Scientific</td><td>BP9744</td><td>Used to pour plates for streaking frozen stocks.</td></tr><tr><td>Air compressor (Jun-Air)</td><td>Elveflow</td><td>KCP-110</td><td>Used to pressurize flow controller.</td></tr><tr><td>Ammonium sulfate ((NH4)2SO4, CAS: 7783-20-2)</td><td>Sigma-Aldrich</td><td>A4915</td><td>Used for experimental media and solutions.</td></tr><tr><td>Angled tweezers</td><td>TDI International</td><td>7-SA</td><td>Used to attach tubing to microfluidic reactor.</td></tr><tr><td>AutoCAD software</td><td>Autodesk&nbsp;</td><td>--</td><td>Used to design microfluidic reactor geometry.</td></tr><tr><td>Autoclave</td><td>Steris</td><td>--</td><td>Used to sterilize materials and liquids.&nbsp;</td></tr><tr><td>Balance</td><td>Mettler Toledo</td><td>ME2002T</td><td>Used to weigh PDMS elastomer base and curing agent.</td></tr><tr><td>Barbed adapter (1/4-28 swivel to 3/32-in barbed)</td><td>Elveflow</td><td>KFI-06</td><td>Used to connect polyether polyurethane tubing to manifold and fluid reservoir rack.</td></tr><tr><td>Biopsy punch (Rapid-Core, 1.2-mm diameter)</td><td>Darwin Microfluidics</td><td>PT-T983-12-25</td><td>Used to punch inlet/outlet holes in PDMS microfluidic channels.&nbsp;</td></tr><tr><td>Bleach (Chlorox)</td><td>ULINE</td><td>S-19719</td><td>Used to sterilize flow control system. Dilute to 1% in ultrapure water.</td></tr><tr><td>Blunt-tip needle (Jensen Global, 21-gauge)</td><td>QSource</td><td>JG21-10HPX-J004</td><td>Used to connect tubing to syringes. Sterilized by autoclaving for 30 min.</td></tr><tr><td>Bunsen burner</td><td>Humboldt</td><td>H-5970</td><td>Used to maintain a sterile experimental workspace.</td></tr><tr><td>Calcium chloride dihydrate (CaCl2&middot;2H2O, Fisher Chemical, CAS: 10035-04-8)</td><td>Fisher Scientific</td><td>C79</td><td>Used for experimental media and solutions.</td></tr><tr><td>Casamino acid (Fisher BioReagents, acid casein peptone)</td><td>Fisher Scientific</td><td>BP1424</td><td>Used for experimental media.</td></tr><tr><td>Cell density meter (WPA Biowave CO8000)</td><td>Avantor</td><td>490005-906</td><td>Used to measure OD600 of bacterial cultures.&nbsp;</td></tr><tr><td>Clear tape (Scotch Magic)</td><td>ULINE</td><td>S-10223</td><td>Used to keep dust from sticking to PDMS microfluidic channels.</td></tr><tr><td>Cobalt(II) chloride hexahydrate (CoCl2&middot;6H2O, CAS: 7791-13-1)</td><td>Sigma-Aldrich</td><td>255599</td><td>Used for experimental media and solutions.</td></tr><tr><td>Compressed nitrogen</td><td>Airgas</td><td>NIUHP300</td><td>Used to dry microfluidic reactor components.</td></tr><tr><td>Conical tube (Falcon, 15-mL volume, polypropylene)</td><td>Fisher Scientific</td><td>14-959-49B</td><td>Used for influent solutions and ICP-MS samples. Pre-sterilized.</td></tr><tr><td>Conical tube (Falcon, 50-mL volume, polypropylene)</td><td>Fisher Scientific</td><td>14-432-22</td><td>Used for LB preculture. Pre-sterilized.</td></tr><tr><td>Copper(II) sulfate pentahydrate (CuSO4&middot;5H2O, CAS: 7758-99-8)</td><td>Sigma-Aldrich</td><td>C8027</td><td>Used for experimental media and solutions.</td></tr><tr><td>Cutting mat</td><td>ULINE</td><td>S-18543</td><td>Used to cut apart PDMS microfluidic channels and punch inlet/outlet holes.</td></tr><tr><td>Cuvette (Fisherbrand, 1.5-mL volume, polystyrene)</td><td>Fisher Scientific&nbsp;</td><td>14-955-127</td><td>Used to measure OD600 of bacterial cultures.&nbsp;</td></tr><tr><td>Digital camera (Hamamatsu ORCA-Fusion)</td><td>Nikon</td><td>77054115</td><td>Used for time-lapse color brightfield imaging of microfluidic experiments.</td></tr><tr><td>Distribution valve (MUX)</td><td>Elveflow</td><td>MUX-D-12</td><td>Used to sequentially inject multiple fluids.</td></tr><tr><td>Elveflow Smart Interface (ESI) software</td><td>Elveflow&nbsp;</td><td>--</td><td>Used to control flow rate/pressure feedback between flow controller and flow sensor and perform distribution valve switching.</td></tr><tr><td>Erlenmeyer flask (Fisherbrand, 50-mL volume)</td><td>Fisher Scientific</td><td>FB50050</td><td>Used for mid-exponential culture. Sterilized by autoclaving for 30 min.</td></tr><tr><td>Ethanol (Fisher Chemical, reagent alcohol, 90% purity, CAS: 64-17-5)</td><td>Fisher Scientific</td><td>A962P-4</td><td>Used to clean PDMS and cover slips for reactor fabrication. Diluted to 70% in ultrapure water.</td></tr><tr><td>Ethylenediaminetetraacetic acid (EDTA, CAS: 60-00-4)</td><td>Sigma-Aldrich</td><td>EDS</td><td>Used for experimental media and solutions.</td></tr><tr><td>Flangeless fitting (1/4-28 for 1/16-in OD tubing)</td><td>Elveflow</td><td>KFI-17</td><td>Used to connect tubing to Elveflow hardware components. Kit includes ferrules and threaded nuts.</td></tr><tr><td>Flow controller (OB1 MK3+)</td><td>Elveflow</td><td>OB1-2000</td><td>Used to provide flow rate/pressure feedback for controlled fluid injection.</td></tr><tr><td>Flow sensor (MFS2)</td><td>Elveflow</td><td>MFS-D-2</td><td>Used to maintain stable flow for microfluidic experiments using pressure feedback from flow controller.</td></tr><tr><td>Fluid reservoir rack&nbsp;</td><td>Elveflow</td><td>KPT-4S</td><td>Used to pressurize influent fluids. Compatible with 15-mL Falcon conical tubes.</td></tr><tr><td>Glass cover slip (1.42-in x 2.36-in, #1 thickness)</td><td>Ted Pella</td><td>260460-100</td><td>Plasma bonded to PDMS channel as a substrate for microfluidic reactors.&nbsp;</td></tr><tr><td>Glass gas-tight syringe (5-mL volume, Luer-Lock)</td><td>Hamilton</td><td>81520</td><td>Used for reactor inoculation and acid digestion (separate syringes). Inoculation syringe sterilized by autoclaving for 30 min.</td></tr><tr><td>HEPES buffer (CAS: 7365-45-9)</td><td>Sigma-Aldrich</td><td>H3375</td><td>Used for experimental media and solutions.</td></tr><tr><td>Hot plate</td><td>Corning</td><td>6795-420D</td><td>Used to cure PDMS.</td></tr><tr><td>Hydrophobic filter (Elveflow, 0.2-&micro;m)</td><td>Darwin Microfluidics</td><td>LVF-KXX-14</td><td>Used to prevent liquid backflow into the flow controller.</td></tr><tr><td>Inductively coupled plasma mass spectrometer (ICP-MS, 7900)</td><td>Agilent</td><td>--</td><td>Used to measure Mn concentration in microfluidic reactor digest.</td></tr><tr><td>Inverted epifluorescence microscope (Nikon Ti2 Eclipse)</td><td>Nikon</td><td>MEA54000, MEE59925, MEL51920, MEP59394, MEC56130</td><td>Used for time-lapse color brightfield imaging of microfluidic experiments. Basic setup includes body, diascopic pillar, condenser turret, nosepiece, and stage.

uma plataforma microfluídica para investigar a precipitação microbiana de óxidos metálicos em meios porosos

A Microfluidic Platform to Investigate Microbial Precipitation of Metal Oxides in Porous Media

Uma plataforma microfluídica para investigar a precipitação microbiana de óxidos metálicos em meios porosos

Microorganisms commonly precipitate metal oxide nanoparticles in soils, sediments, and other environmentally relevant porous media. However, the formation ...

We've developed a microfluidic platform to study the dynamic precipitation of metal oxides by bacteria in a porous environment. This protocol enables in situ and real-time measurement of mineral precipitation, unlike conventional methods such as batch or column systems. To begin, add 10 milliliters of LB medium to a 50 milliliter conical tube.Inoculate the medium using a single colony of Pseudomonas putida GB-1, obtained from a plate that was streaked 20 hours earlier. Incubate the LB pre-culture in a shaker incubator at 30 degrees Celsius and 180 revolutions per minute for 16 hours to reach the stationary phase. Then, pellet one milliliter of LB pre-culture by centrifugation at 8, 000 G for one minute.After washing the pellet three times, re-suspend the final pellet in one milliliter of salt solution without manganese. Then, dilute the washed cell suspension fivefold in salt solution without manganese in a cuvette, and measure the optical density at 600 nanometers using a cell density meter. Inoculate 20 milliliters of growth medium without manganese with the washed cell suspension to an optical density of 0.01 in a 50 milliliter Erlenmeyer flask.Incubate at 30 degrees Celsius and 180 revolutions per minute for six hours to reach mid-exponential phase. Before beginning the experiment, verify that the microscope incubator is maintained at 30 degrees Celsius and prime the flow control system. Pour 10 milliliters of growth medium and salt solution with manganese into a 15 milliliter conical tube, and connect both tubes to the fluid reservoir rack.Degas the prepared microfluidic reactor in a vacuum desiccator for 20 minutes. Draw one to two milliliters of salt solution with manganese into a three milliliter plastic syringe. Attach a blunt tip needle and 20 centimeters of 0.02 inch inner diameter Tygon tubing, press the syringe plunger until the tubing is fully saturated.After de-gassing, use angled tweezers to attach the tubing to the reactor outlet hole. Manually saturate the reactor with the syringe until a droplet forms at the reactor inlet hole. Next, attach 20 centimeters of 0.02 inch inner diameter Tygon tubing to the reactor inlet hole.Continue dispensing salt solution with manganese into the reactor until all air bubbles are removed. To prepare the reactor inoculum, remove the mid-exponential culture from the shaker incubator six hours after inoculation. Dilute fourfold in salt solution without manganese in a cuvette.And measure the optical density. Then, dilute the mid-exponential culture in two milliliters of salt solution with manganese to an initial optical density of 0.005 in a two milliliter microcentrifuge tube. Invert the tube several times to mix.Next, place the reactor inlet tubing in the microcentrifuge tube, ensuring that a droplet of fluid contacts the inoculum to prevent air introduction in the tubing. Partially close the microcentrifuge tube and secure the lid with tape to prevent contamination. Next, withdraw four to five milliliters of salt solution with manganese into a five milliliter glass gas tight syringe to wet the barrel.Dispense the solution until 0.5 milliliters remain in the syringe and load the syringe into a syringe pump. After disconnecting the plastic syringe from the needle, attach the needle to the glass syringe while avoiding trapping air in the needle hub. To inoculate the reactor, withdraw the inoculum into the reactor for 20 minutes at a flow rate of 8.33 microliters per minute.Turn off the syringe pump following inoculation and allow the bacteria to settle and attach for one hour with no flow. Then, remove the glass syringe from the syringe pump while leaving it connected to the outlet tubing. Next, move the reactor and syringe from the bench top to the microscope incubator.On the microscope stage, remove the inlet tubing from the reactor and push gently on the syringe plunger to create a droplet at the reactor inlet hole. Gently bend the polytetrafluoroethylene, or PTFE tubing, from the flow control system at a right angle. Dip the end of the PTFE tubing into a 1%polyethylene glycol diacrylate solution to wet the exterior of the hydrophobic tubing.Ensure a droplet of growth medium with manganese is present at the end of the PTFE tubing, and attach the tubing to the reactor inlet hole. After disconnecting the syringe, place the outlet tubing into a five milliliter microcentrifuge tube to collect the reactor effluent. Set the flow rate to 1.33 microliters per minute in the flow control system software.Program the system to supply growth medium with manganese for 15 hours to promote biofilm formation, followed by salt solution with manganese for 28.5 hours to induce manganese biomineralization. Next, place a sample frame underneath the reactor for stability. Rigidly mount the frame and reactor in the microscope stage insert to ensure full immobilization.Affix the PTFE tubing to the stage insert with a piece of tape to prevent its detachment from the reactor inlet hole. Align the ultraviolet C-arms two millimeters from the pore space edge on the outlet side and three millimeters on the inlet side to minimize UV light scattering and biofilm damage. Set up a program for reactor irradiation to treat the reactor inlet and outlet regions for five minutes at the experiment onset, then treat only the outlet region for five minutes, 17.5 hours after the experiment onset to inactivate remaining biofilms.Manually adjust the Kohler alignment, following the manufacturer's instructions to ensure even illumination. To prevent quilting artifacts, select Acquire and open the shading correction panel, then capture a shading image and set it as the current shading image. Select ND Acquisition and choose XY to set the center coordinate of the stitched image.Next, choose Large Image and set up the stitched image scan area as five by four fields of view, and apply a 1%overlap. Select Lambda and set up the acquisition of Color Brightfield images using the 4X objective. Select time and set the imaging frequency with intervals of one hour for 15 hours and 0.5 hours for 28.5 hours, then run the time lapse program.Brightfield images demonstrated that Pseudomonas putida GB-1 biofilms were uniformly distributed throughout the microfluidic pore space at 18.5 hours before the onset of manganese oxide precipitation. Subsequently, manganese oxides preferentially accumulated on the inlet side of the pore space closest to the influent manganese source. The difference between the reference image and subsequent image time points revealed the contribution of manganese oxide precipitates to the overall image intensity.Quantification of the observed mineral accumulation gradient was enabled at the reactor scale. At the biofilm scale, the image subtraction approach captured the gradual precipitation of manganese oxides on biofilm surfaces and accumulation of minerals at the edges of biofilms in contact with pore fluid. The resulting estimated rate of manganese oxide precipitation was non-monotonic, increasing steeply between 19.5 hours and 21 hours, peaking at 0.29 micrograms per hour and decelerating gradually until 43.5 hours.This protocol allows researchers to study the timing and location of mineral precipitation in a porous medium. It's critical to prevent biofilm growth in the reactor inlet region. This ensures a constant boundary condition and restricts precipitation to the pore space.Future studies can explore coupled microbe mineral fluid interactions under conditions relevant for environmental and engineering applications.

a-microfluidic-platform-to-investigate-microbial-precipitation-metal

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JoVE Journal

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Dispositivos microfluídicos pode ser usado para visualizar os complexos processos naturais em tempo real e nas escalas físico apropriado. Temos desenvolvido um dispositivo simples microfluídicos que imita características-chave de meios porosos naturais para o estudo de crescimento e de transporte de bactérias no subsolo.

Janela em um Microworld: Simple sistemas microfluídicos para estudar o transporte microbiana em Meios Porosos

Gota plataformas baseadas microfluídicos são candidatos promissores para a experimentação de alto rendimento, uma vez que são capazes de gerar picolitros, auto-compartimentada vasos barata a taxas kHz. Através da integração com rápido, sensível e de alta resolução de fluorescência métodos espectroscópicos, a grande quantidade de informações geradas nesses sistemas podem ser eficientemente extraído, aproveitada e utilizada.

Métodos de detecção de fluorescência para as plataformas de gotículas microfluídicos

Fonte: Kurz, D. L., et al. Plataforma microfluídica para estudar bioentupimento em meios porosos. J. Vis. Exp. (2022)O vídeo demonstra o uso de um dispositivo microfluídico para estudar a formação de biofilmes. Uma suspensão bacteriana é introduzida no microcanal, seguida de incubação para permitir o desenvolvimento do biofilme. O fluxo é então retomado, e o monitoramento de pressão junto com imagens é usado para detectar formação de biofilme e obstrução dos poros ao longo do tempo.

Um dispositivo microfluídico para imagem em tempo real e acompanhamento de pressão durante a formação do biofilme

Este protocolo descreve uma arquitetura de sistema para realizar separações automatizadas de partículas de pequeno volume (0,15–1,5 ml) usando um dispositivo microfluídico e discute métodos para otimizar o desempenho e a operação do dispositivo acustofluídico.

Uma plataforma microfluídica para processamento de precisão de pequeno volume de amostra e respectiva utilização para Tamanho partículas biológicas separadas com um microdispositivos Acústico

Sistemas de laboratório-em-um-gota reação permitem a implementação versátil de reações complexas em uma escala de microfluidic. Uma plataforma de acionamento automatizado que consiste de uma matriz 3 x 3 das bobinas electromagnéticas foi desenvolvida e utilizado com sucesso para mesclar dois 10 µ l de microreactors e, assim, iniciar uma reação enzimática em mármores do líquido resultantes.

Aquosas gotículas usadas como Microreactors enzimática e sua atuação eletromagnética

Uma plataforma de biosensor microfluidic foi projetada e fabricado usando a tecnologia de fotorresiste de película seca de baixo custo para a quantificação rápida e sensível de vários analitos. Este sistema de uso único permite a leitura eletroquímica de ensaios de enzima-lig na-microplaqueta-imobilizado mediante a técnica de stop-fluxo.

Película seca baseada em fotorresiste eletroquímica Microfluidic Biosensor plataforma: Fabricação de dispositivo, ensaio em-microplaqueta preparação e operação de sistema

O objetivo deste procedimento é rapidamente e facilmente produzir um dispositivo microfluidic com geometria personalizável e resistência ao inchaço por fluidos orgânicos para estudos de recuperação de óleo. Um molde de polidimetilsiloxano é gerado pela primeira vez e então usado para converter o dispositivo baseado em epóxi. Um estudo representativo do deslocamento é relatado.

Dispositivos microfluídicos para caracterizar processos de evento Pore-escala em meios porosos para aplicações de recuperação de óleo

Detalhadas neste documento são os protocolos de operação e montagem de uma plataforma de triagem microfluídicos modular para a caracterização sistemática das sínteses de nanocrystal semicondutor coloidal. Através de dispositivos de sistema totalmente ajustável, coleção de espectros altamente eficiente pode ser efectuada através de escalas de tempo de reação de 4 ordens de grandeza dentro de um espaço de amostragem massa transferência controlada.

Uma tecnologia Microfluidic Modular para estudos sistemáticos de nanocristais semicondutores coloidal

Apresentamos um sistema microfluido para estudos de alto rendimento em máquinas de vida complexas, que consiste em 1500 unidades culturais, uma matriz de bombas peristálticas aprimoradas e um módulo de mistura no local. O chip microfluido permite a análise das condições microambientais altamente complexas e dinâmicas in vivo.

Geração de Condições Ambientais Dinâmicas usando um dispositivo microfluido de alto rendimento

O presente protocolo descreve uma plataforma microfluidica pneumática que pode ser usada para uma concentração eficiente de micropartículas.

Plataforma microfluídica pneumática para concentração de micropartículas

O presente protocolo descreve uma plataforma microfluídica para estudar o desenvolvimento de biofilme em meios porosos quase-2D, combinando imagens de microscopia de alta resolução com medidas simultâneas de diferença de pressão. A plataforma quantifica a influência do tamanho dos poros e das taxas de fluxo de fluido em meios porosos no bioentupimento.

Uma plataforma microfluídica para o estudo do bioentupimento em meios porosos

University of California, Davis.  Este protocolo apresenta um sistema microfluídico para estudar a precipitação microbiana dos minerais de óxido de manganês. A formação de biominerais é medida in situ por microscopia óptica e análise de imagem. Essa abordagem pode ser adaptada para investigar a precipitação de outros minerais biogênicos de cores distintas em ambientes porosos.

Uma plataforma microfluídica para investigar a precipitação microbiana de óxidos metálicos em meios porosos

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