About 0.2 g samples from 10 replicates of each FS were diluted separately in 10 mL of filter-sterilized, surfactant-free ultra-pure hot water (Additional file 1 : Figure S9) and subsequently filtered through 20–25 μm (Grade 2 Whatman®), 2.5 μm (Grade 41 Whatman®), 0.4 μm (Whatman® 110607, Nuclepore membrane filter) and 0.2 μm (Whatman® 7182-014, Cellulose nitrate membrane filter). Final filtrates obtained from a 0.2 μm filter were pooled together with the respective FS and quantified. The spectral and morphological features of the dried NPs were analysed using Fourier Transform Raman Spectroscopy (FT-Raman) (PerkinElmer 1600 instrument, USA) and HR-SEM (Carl Zeiss Evo 18 SEM, Germany). The dispersion, stability, and size distribution of the NPs in culture medium (DMEM) were measured using the Malvern Dynamic Light Scattering instrument (Malvern, UK) in triplicates with 20 runs each.
Nuclepore membrane filter
Manufactured by Cytiva
Sourced in United States
The Nuclepore membrane filter is a laboratory equipment used for filtration purposes. It is a thin, porous membrane made of polycarbonate material, designed to remove small particles or molecules from liquid or gas samples. The filter features precisely controlled pore sizes, providing a reliable and consistent filtration process.
Automatically generated - may contain errors
Lab products found in correlation
Dnase 1, by Thermo Fisher Scientific (2 mentions)
Dynamic light scattering instrument, by Malvern Panalytical (1 mentions)
Evo 18 sem, by Zeiss (1 mentions)
Sterivex cartridge filter, by Merck Group (1 mentions)
Bromodeoxyuridine (brdu), by Merck Group (1 mentions)
Vp 30s, by TAITEC (1 mentions)
Nuclepore filter, by Cytiva (1 mentions)
Rlt buffer, by Qiagen (1 mentions)
Primescript rt reagent kit with gdna eraser, by Takara Bio (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Lysozyme, by Cytiva (1 mentions)
Milli q integral 3 water production unit, by Merck Group (1 mentions)
Round mica sheets, by Ted Pella (1 mentions)
Sm2 bio beads, by Bio-Rad (1 mentions)
1 2 dimyristoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
8 protocols using nuclepore membrane filter
1
Characterization of Nanoparticles from Functional Substrates
2
Bacterial Cell Isolation from Seawater and Sediment
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Approximately 10 L of seawater samples and 50 mL of resuspendable sediment samples were incubated with BrdU (1 μM final concentration; Sigma-Aldrich Co. LLC) at ambient water temperatures (Table 1 ) for 5 h. After incubation, the bacterial cells in the seawater samples were collected with a Nuclepore™ membrane filter (3.0 μm pore size, 47 mm diameter; Whatman) and subsequently with a Sterivex™ cartridge filter (0.22 μm pore size; Millipore, MA, USA) using a peristaltic pump. The 3.0 μm Nuclepore™ membrane filters were changed 2–3 times before the filters became clogged. The bacterial cells in the sediment samples were collected with a Sterivex™ cartridge filter (0.22 μm pore size) using a syringe. Immediately after filtration, the filters were stored at −20°C until further analysis.
3
Size-Fractionated Chlorophyll a Quantification
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Seawater samples for small (0.7–2.0 μm) and large (>2.0 μm) size-fractionated chlorophyll a (chl a) molecules were obtained from the three depths at which light was measured (i.e., 100, 30, and 1% PAR penetration). To estimate the size composition of the phytoplankton assemblages, the seawater samples were passed sequentially through a 2 μm (large chl a) Nuclepore membrane filter (47 mm) and then a 0.7 μm (small chl a) Whatman GF/F paper (47 mm). The filters were frozen immediately for further analysis in the laboratory. After extraction in 90% acetone, the concentrations of the size-fractionated chl a were determined with a previously calibrated fluorometer (Turner Designs model 10-AU) based on methods described by Kim et al. (2015) (link).
4
Extraction and Analysis of MWCNT-Adsorbed B[ghi]P
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Paraformaldehyde fixed lung samples were allowed to react with Clean 99‐K200R (C99) (Clean Chemical, Osaka, Japan) at room temperature overnight. The solution was then centrifuged at 13 000 × g for 10 min and the supernatant was removed. The precipitate was resuspended in 1 mL of 9.6% PBS containing 0.1% Tween 80 (TW‐solution) followed by a second centrifugation at 13 000 × g for 10 min. The pellet was resuspended in 100‐μL concentrated sulfuric acid to remove the organic content. 1‐mL TW‐solution was stirred into the acid‐MWCNT‐N mixture. 25 μL of a solution of 0.125 μg/mL Benzo[ghi]perylene (B[ghi]P) in acetonitrile was then added, and the B[ghi]P was allowed to adsorb onto the MWCNT for 15 min. The mixture was passed through a Nuclepore membrane filter (Whatman 111109, pore size 0.8 μm, diameter 47 mm; Fisher Scientific, Waltham MA, USA), and the MWCNT containing region was punched out and placed in 1‐mL acetonitrile and sonicated for 10 s with an ultrasonic homogenizer (VP‐30S, 20 kHz, 300 W; TAITEC, Tokyo, Japan). The solution was then filtered for HPLC analysis of the extracted B[ghi]P.
5
Diatom Gene Expression Profiling
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Samples for DNA and RNA analyses were collected on days 0, 2 and 3. DNA samples (300–400 mL) were collected onto 0.2 μm pore size polycarbonate Nuclepore membrane filters (Whatman) with gentle vacuum (< 0.013 MPa), flash frozen in liquid nitrogen and then stored in a deep freezer at −80°C until analysis. Seawater samples (300–400 mL) for RNA analysis were filtered onto 0.2 μm pore size polycarbonate Nuclepore filters (Whatman) with gentle vacuum (< 0.013 MPa) and then stored in 1.5-mL cryotubes previously filled with 0.2 g of muffled 0.1-mm glass beads and 600 μL of RLT buffer (Qiagen) with 10 μL mL−1 β-mercaptoethanol (Sigma-Aldrich). The RNA samples were flash-frozen in liquid nitrogen and stored in a deep freezer at −80°C until analysis. The detailed methodologies of total DNA and RNA extractions were described in Endo et al. [16 ] and Endo et al. [27 ], respectively. The extracted RNA was reverse-transcribed into cDNA using the PrimeScript RT Reagent Kit with gDNA Eraser (Takara). To quantify the diatom-specific rbcL gene (DNA) and its transcripts (cDNA), qPCR and qRT-PCR were performed according to the method of Endo et al. [27 ].
6
DMPC Lipid Membrane Preparation
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1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) was from Avanti Polar Lipids Inc. (Alabaster, AL). Lipopolysaccharides (LPS, rough strains) from Escherichia coli EH100 (Ra mutant), Whatman Nuclepore membrane filters (track-etched polycarbonate membranes, pore diameter 100 nm), octyl glucopyranoside (OG), sarkosyl, poly-l -lysine (PLL) (150 000–300 000 g mol−1), lysozyme, sodium azide, calcium, Trizma base and 1 M magnesium chloride solutions were from Sigma-Aldrich. DNAse I was from Thermo Fisher Scientific. Argon 5.0 and nitrogen 5.0 gases were from Linde Gáz Magyarország Zrt (Budapest, Hungary). Water was purified with a Milli-Q Integral 3 Water Production Unit (Merck Millipore, Billerica, MA). Round mica sheets were from Ted Pella, Inc. (Redding, CA). Detergent adsorbent SM-2 Bio Beads were from Bio-Rad.
7
Filtered Rainwater Analysis of Nitrogen Ions
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Immediately after the collection, throughfall and rainfall samples were sequentially filtered to remove suspended solids (SS) and microbes using Nuclepore membrane filters (Whatman, Middlesex, UK) with pore sizes of 10 (to remove large SS and aggregated microbes), 2 (to remove medium SS and large microbes), 0.4 (to remove small SS and most microbes), and 0.2 µm (to remove very small SS and large viruses) (Fig. 1 ). All unfiltered and filtered samples (50 mL in each fraction) were dispensed into sterile 50-mL Erlenmeyer flasks with foam silicon stoppers and incubated at 30 °C in the dark. Following incubation periods of 1, 2, 3, and 4 weeks, nitrate, nitrite, and ammonium ions in each sample were analyzed using ion chromatography (DX-100) with an IonPack AS12A/AG12A and IonPack CS3/CG3 column (Dionex, Sunnyvale, CA, USA). Analysis of ammonium ions was only conducted for samples before incubation and after incubation for 1 and 4 weeks. The filtrates and filters were stored at −20 °C until further analysis.
8
Seawater CDOM Optical Density Measurement
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All laboratories filtered replicate seawater samples through 0.2 μm Whatman Nuclepore membrane filters into acid cleaned glassware. The first two 0.25 L of the filtered seawater were discarded. a CDOM (λ) was determined using the third sample in a 10 cm quartz cuvette from 350 to 750 nm relative to a bi-distilled MilliQ reference blank and was calculated from the optical density of the sample and the cuvette path length following the protocols given in Tilstone et al. (2004) .
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