Abstract
Les biofilms sont des communautés microbiennes attachée surface qui ont des structures complexes et produisent hétérogénéités spatiales importantes. le développement du biofilm est fortement réglementé par le flux environnante et l'environnement nutritionnel. la croissance du biofilm augmente également l'hétérogénéité du microenvironnement local en générant des champs d'écoulement complexes et des structures de transport de solutés. Pour étudier le développement de l'hétérogénéité dans les biofilms et les interactions entre les biofilms et leur micro-habitat local, nous avons grandi biofilms mono-espèces de Pseudomonas aeruginosa et les biofilms double-espèces de P. aeruginosa et Escherichia coli sous gradients nutritionnels dans une cellule d'écoulement microfluidique. Nous fournissons des protocoles détaillés pour la création de gradients de nutriments dans la cellule d'écoulement et pour la culture et la visualisation de développement du biofilm dans ces conditions. Nous avons également des protocoles actuels pour une série de méthodes optiques de quantifier la répartition spatiale de la structure du biofilm, distri circulerbutions sur les biofilms, et le transport de masse autour et à l'intérieur des colonies de biofilm. Ces méthodes prennent en charge des enquêtes approfondies de la co-développement du biofilm et l'habitat hétérogénéité.
Materials
| Name | Company | Catalog Number | Comments |
| Peristaltic Pump | Gilson | Miniplus 3 | Flow cell setup and inoculation |
| Pump Tubing 0.50 mm OVC, Orange/Yellow | Gilson | F117934 | Flow cell setup and inoculation |
| Three-way Stopcock w/ Swivel Male Luer lock | Smiths Medical | MX9311L | Flow cell setup and inoculation |
| Sylgard 184 Solar Cell Encapsulation for Making Solar Panels | ML Solar LLC | Flow cell setup and inoculation | |
| Pyrex Medium Bottle, 1 L, GL45 | VWR | 16157-191 | Flow cell setup and inoculation |
| C-FLEX Tubing | Cole-Parmer | 06422-02 | Flow cell setup and inoculation |
| 1 ml TB Syringe | BD | 309659 | Flow cell setup and inoculation |
| Polymer Tubing | IDEX | 1520G | Flow cell setup and inoculation |
| Sterile Intramedic Luer Stub Adapter | Clay Adams | 427564 | Flow cell setup and inoculation |
| PrecisionGlide Needle | BD | 305195 | Flow cell setup and inoculation |
| Spectrophotometer | HACH | Flow cell setup and inoculation | |
| Syringe filters - sterile (0.2 μm) | Fisherbrand | 09-719A | Flow cell setup and inoculation |
| MAXQ Shaker | Thermo Scientific | Flow cell setup and inoculation | |
| Ammonium sulfate | Sigma Aldrich | A4418 | Growth media |
| Sodium phosphate dibasic anhydrous | Sigma Aldrich | RES20908-A7 | Growth media |
| Monobasic potassium phosphate | Sigma Aldrich | P5655 | Growth media |
| Sodium chloride | Sigma Aldrich | S7653 | Growth media |
| Magnisium chloride | Sigma Aldrich | M8266 | Growth media |
| Calcium chloride | Sigma Aldrich | C5670 | Growth media |
| Calcium sulfate dihydrate | Sigma Aldrich | C3771 | Growth media |
| Iron(II) sulfate heptahydrate | Sigma Aldrich | 215422 | Growth media |
| Manganese(II) sulfate monohydrate | Sigma Aldrich | M7634 | Growth media |
| Copper(II) sulfate | Sigma Aldrich | 451657 | Growth media |
| Zinc sulfate heptahydrate | Sigma Aldrich | Z0251 | Growth media |
| Cobalt(II) sulfate heptahydrate | Sigma Aldrich | C6768 | Growth media |
| Sodium molybdate | Sigma Aldrich | 243655 | Growth media |
| Boric acid | Sigma Aldrich | B6768 | Growth media |
| Dextrose | Sigma Aldrich | D9434 | Growth media |
| Luria Bertani Broth | Sigma Aldrich | L3022 | Growth media |
| TCS SP2 Confocal Microscopy | Leica | Fluorescent imaging | |
| SYTO 62 | Life Technology | S11344 | Fluorescent imaging |
| Cy5 | GE Healthcare Life Sciences | PA15100 | Fluorescent imaging |
| Red Fluorescent (580/605) FluoSphere | Life Technology | F-8801 | Fluorescent imaging |
| BioSPA | Packman Lab | Image Processing | |
| ImageJ | NIH | Image Processing | |
| Volocity | PerkinElmer | Image Processing | |
| Streams 2.02 | University of Cantebury | Image Processing |
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