Filtration volumes were adjusted between 100 and 5'000 ml per filter based on FCM total cell concentration (TCC) measurements to approximately equalize number of cells captured (Table S2 ). Three types of filters were captured in duplicate for each sample (Fig. 1b ). The first filter captured the entire community with direct filtration onto 0.2 µm membrane filters (“all bacteria”) (NucleporeTM track-etched polycarbonate membranes, 47 mm, Whatman, UK) using sterilized filtration units (NalgeneTM, Thermo Fisher Scientific, USA) mounted on sterilized glass bottles. Separately, a two-step filtration was performed to obtain size-based groups. Another water sample was first filtered onto 0.4 µm membrane filters (large bacteria) (NucleporeTM track-etched polycarbonate membranes, 47 mm, Whatman, UK), and the resulting filtrate was subsequently filtered again on 0.2 µm filters (small bacteria). Filters from the paired filtration step (Large, small) and direct filtration (All) were then stored at −80 °C until DNA extraction.
Nuclepore track etched polycarbonate membrane
Manufactured by Cytiva
Sourced in United Kingdom
Nuclepore Track-Etched polycarbonate membranes are a type of lab equipment used for filtration and separation applications. They feature a defined and uniform pore structure created through a track-etching process. These membranes provide precise control over pore size and distribution, enabling efficient filtration and analysis of various samples.
Automatically generated - may contain errors
Lab products found in correlation
Nalgene, by Thermo Fisher Scientific (1 mentions)
Avanti mini extruder, by Merck Group (1 mentions)
1 4 dioxane, by Carl Roth (1 mentions)
Tetrahydrofuran thf, by Carl Roth (1 mentions)
Type 100 qs, by Hellma (1 mentions)
Ps b paa, by Merck Group (1 mentions)
Avanti s mini extruder, by Avanti Polar Lipids (1 mentions)
7 protocols using nuclepore track etched polycarbonate membrane
1
Size-based Bacterial Community Filtration
2
Preparation of Lipid Vesicles by Extrusion
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The lipids used in the present study are listed in Table 4 . The lipids were dissolved in chloroform, flushed with liquid nitrogen, and kept in glass flasks at –20°C. For each experiment, 400 nmol of the respective lipid mixture or, in case of complex lipid mixtures such as total brain lipids, 500 μg of the lipid mixture was dried under nitrogen gas in a 2-mL round-bottom plastic tube and rehydrated in 25 μL of hydration buffer (as indicated below) for 1 h in a 60°C water bath. An Avanti Mini Extruder (Sigma-Aldrich, catalog no. 610020-1EA) was assembled. The turbid lipid mixture was ultrasonicated for 90 s (Branson Ultrasonic CL-40549) and filled into an extruder-compatible 1-mL Hamilton syringe (Sigma-Aldrich, catalog no. 610017-1EA). Whatman Nuclepore Track-Etched polycarbonate membranes (pore size, 0.2 μm) were inserted. The lipid mixture was equilibrated on the Mini Extruder heating bloc (Sigma-Aldrich, catalog no. 610024) for 10 min at 60°C and then pressed 21 times through the extruder. The resulting liposomes were stored at 4°C until use, with a maximum of 5 days.
3
Quantifying Pseudomonas aeruginosa Biofilm
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Bacterial cultures in BHI (with 50 mM KNO3) were grown to the log phase (OD600nm around 0.5) and then diluted to OD600nm 0.02 with fresh medium. Next, 10 μl diluted cultures were spotted onto 0.2-μm Nuclepore track-etched polycarbonate membranes (Whatman) that were placed (smooth side facing upwards) on BHI agar plates (with 50 mM KNO3). The plates were then incubated in the anaerobic chamber at 37 °C for 16 h. Membranes containing P. aeruginosa biofilms were transferred to the BeadBug prefilled tubes that contained 1 ml phosphate-buffered saline (PBS). Biofilms were mechanically disrupted by bead beating for 45 s. CFUs were determined by spreading the diluted cell suspensions on PIA plates.
4
Synthesis and Characterization of PS-b-PAA
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Polystyrene-block-polyacrylic acid (PS-b-PAA, batch: MKBQ5839V) was purchased from Sigma Aldrich (Steinheim, Germany). The degree of polymerization of PS is 275 and of PAA 30, as stated by the manufacturer. The molecular weights (Mn) and the polydispersity index (PDI) of PS-b-PAA are summarized in Table 1 .
Deionized water was used as the selective solvent for the hydrophobic part (polystyrene). Tetrahydrofuran (THF, for HPLC, not stabilized) and 1,4-dioxane (for HPLC, not stabilized) from Carl Roth (Karlsruhe, Germany) were utilized as good solvents for PS-b-PAA.
Filters were used for microscopy (Whatman® Nuclepore™ track-etched polycarbonate membranes, with pore diameters of between 0.2 µm and 2.0 µm, Dassel, Germany). A quartz glass cuvette (Hellma Analytics, Type 100 QS, Müllheim, Germany) with a layer thickness of 10 mm and a nominal volume of 3.5 mL was used for dynamic light scattering measurements.
Deionized water was used as the selective solvent for the hydrophobic part (polystyrene). Tetrahydrofuran (THF, for HPLC, not stabilized) and 1,4-dioxane (for HPLC, not stabilized) from Carl Roth (Karlsruhe, Germany) were utilized as good solvents for PS-b-PAA.
Filters were used for microscopy (Whatman® Nuclepore™ track-etched polycarbonate membranes, with pore diameters of between 0.2 µm and 2.0 µm, Dassel, Germany). A quartz glass cuvette (Hellma Analytics, Type 100 QS, Müllheim, Germany) with a layer thickness of 10 mm and a nominal volume of 3.5 mL was used for dynamic light scattering measurements.
5
Chloride Efflux Measurement of EYPC Vesicles
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The 1H and 13C NMR spectra were measured on a Bruker Avance AV 400 or 500 spectrometer, and the data were reported relative to the deuterium solvents. Waters UPLC/Quattro Premier XE and Bruker maXis 4G ESI-Q-TOF mass spectrometers were used to measure the LR and HR ESI-MS spectra, respectively. Analytical thin-layer chromatography (TLC) plates (silica gel, GF254) were detected by use of iodine and UV (254 or 365 nm). EYPC vesicles were prepared by extrusion through nuclepore track-etched polycarbonate membranes (100 nm, Whatman, Florham Park, New Jersey, USA) on an Avanti's Mini-Extruder (Avanti Polar Lipids, Inc., Alabaster, Alabama, USA). Chloride efflux was measured by using a Mettler-Toledo PerfectIon™ chloride ion selective electrode assembled with a Mettler-Toledo Seven Compact S220 ionometer.
EYPC and MTT were purchased from Sigma Chemical Co. (St Louis, USA). All the other chemicals and reagents were obtained from commercial sources and used without further purification. The experimental protocols for the measurement of anion recognition, anion transport and biological activity were included in the ESI.†
EYPC and MTT were purchased from Sigma Chemical Co. (St Louis, USA). All the other chemicals and reagents were obtained from commercial sources and used without further purification. The experimental protocols for the measurement of anion recognition, anion transport and biological activity were included in the ESI.
6
Fabrication of IOIF Nanofiltration Membranes
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The IOIF membranes were prepared by a simple filtration of sample solutions containing single-layer IOIF self-assemblies through a commercial filter with even dispersed pores in certain size under vacuum pressure of −2,000 Pa. After washing with water, the prepared nanofiltration membrane has the same size as the effective filtration area of supporting substrate used in the installation. In a typical procedure, 20 ml as-prepared [Azo-TrEG@CD][PWV] solution (0.04 mg ml−1) was filtered over a supporting filter (Whatman Nuclepore Track-Etched Polycarbonate Membrane; effective filtration area: 3.14 cm2; pore size: 200 nm) under the preset vacuum pressure. Water (10 ml) was subsequently used to wash the membrane. Under the same vacuum pressure, the IOIF membranes with various thicknesses were prepared by using 20 ml sample solution with concentrations: 0.02, 0.04, 0.06 and 0.08 mg ml−1, and corresponding thicknesses are measured to be 0.20∼0.35, 0.43, 1.43 and 2.19 μm, respectively.
7
Preparation of Azo-TrEG@CD Powder and Film
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The powdered sample was prepared by the lyophilization of the [Azo-TrEG@CD][PWV] solution (0.17 mg ml−1, 120 ml) and then grinding it into powder. The film sample was prepared by the filtration of [Azo-TrEG@CD][PWV] solution (0.18 mg ml−1, 120 ml) over a supporting filter (Whatman Nuclepore Track-Etched Polycarbonate Membrane; effective filtration area: 3.14 cm2; pore size: 200 nm), drying in oven at 40 °C for 48 h.
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