Phytoplankton samples were collected from upper 200 m water column by using a phytoplankton net (64 µm mesh), on the Western Pacific Ocean (7°0.26'N, 141°59.63'E LM observation, cleaned samples were mounted on glass slides with Mountmedia (Wako Pure Chemical Industries, Ltd., Osaka, Japan). A Zeiss Imager Z2 microscope (Carl Zeiss Microimaging GmbH, Jena, Germany) with differential interference contrast (DIC ) was used for LM observation. The measurement methods of the raphe angle and the intersection angle of the oblique striae followed Sterrenburg (1991) (link). For SEM observations, specimens were placed on coverslips, air-dried and coated with osmium. A Hitachi S-4800 (Hitachi, Ltd., Tokyo, Japan) was used for SEM observation.
Mountmedia
Manufactured by Fujifilm
Sourced in Japan
Mountmedia is a lab equipment product by Fujifilm. It is designed for mounting and storing media samples. The core function of Mountmedia is to provide a secure and organized way to handle and preserve media specimens for analysis and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Imager z2 microscope, by Zeiss (2 mentions)
S 4800, by Hitachi (2 mentions)
Tetramethylsilane, by Merck Group (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
S 4300 scanning electron microscope, by Hitachi (1 mentions)
Eos m10, by Canon (1 mentions)
Tm3000 tabletop microscope, by Hitachi (1 mentions)
4 protocols using mountmedia
1
Diatom Isolation and Cultivation from Pacific Ocean
2
Diatom Strain Identification Protocol
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Monoclonal cultures of benthic diatom strains were identified to the genus or species level by morphological features based on observations under light and scanning electron microscopy. For the light microscopy examination, diatom cultures were treated with acid to prepare cleaned frustules [30 ], and then permanent slides were made using Mountmedia (Wako Pure Chemical Industries, Osaka, Japan). The slides were examined using light microscopy under a ×100 oil immersion objective lens (Eclipse 80i; Nikon). For scanning electron microscopy examination, diatom cells fixed with Lugol’s solution were filtered onto a polycarbonate filter (diameter of 25 mm; pore size of 1 or 2 μm) and then washed with distilled water. The filter papers were dehydrated in a graded ethanol series (10%, 25%, 50%, 75%, 90%, and 100%) and dried using tetramethylsilane (Sigma-Aldrich, St. Louis, MO, USA). Finally, the samples were mounted onto stubs and sputter-coated with platinum. Observations were performed with a Hitachi S–4300 scanning electron microscope (Hitachi, Tokyo, Japan). The previous studies were referred to for instructions on morphological comparisons [31 –41 ]. Strains that did not match those in the published literature were treated as unidentified species.
3
Microscopic Examination of Cultured Cells
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After cultured cells were observed using a light microscope, some cells were treated with alkaline detergent, washed with distilled water, embedded in Mount Media (Fujifilm Wako, Osaka) and observed using a light microscope again. Bright-field images were photographed using a CX23 Biological Microscope (Olympus, Tokyo) equipped with digital camera EOS M10 (Canon, Tokyo). For scanning electron microscopy, specimens after cleaning were coated with gold particles in an Ion Coater IB-3 (Eiko, Tokyo), with an ionization current of 5 mA for 3 min. Scanning electron microscopic images were taken using a Tabletop Microscope TM3000 (Hitachi, Tokyo).
4
Isolation and Cultivation of Benthic Diatoms
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Surface muddy sediments with benthic diatoms were sucked up by using a glass tube from an intertidal sand beach in the Huiquan Bay, Qingdao City. The diatom cells were isolated using capillary pipettes and cultivated in 250 mL cell culture flasks with 100 mL F/2 medium. Cultures were maintained at 20–22 °C under a low light intensity (25–30 µmol photo/m2/s) with a light/dark cycle of 12:12 h. Five millilitres of diatom culture were fixed with 2.5% glutaraldehyde and then cleaned with hydrogen peroxide to remove organic components of frustules [33 (link)].
Morphological observation followed the method described in our previous study [34 (link)]: for light microscopy (LM) observation, cleaned samples were pipetted onto the coverslips, air-dried, and mounted on glass slides with Mountmedia (Wako Pure Chemical Industries, Ltd., Osaka, Japan). LM microphotographs of cleaned frustules were taken by using a Zeiss Imager Z2 microscope (Carl Zeiss Microimaging GmbH, Jena, Germany) with differential interference contrast (DIC). For scanning electron microscopy (SEM) observation, cleaned frustules were placed on round coverslips, air-dried, and coated with osmium. SEM observation was performed by using a Hitachi S–4800 (Hitachi, Ltd., Tokyo, Japan).
Morphological observation followed the method described in our previous study [34 (link)]: for light microscopy (LM) observation, cleaned samples were pipetted onto the coverslips, air-dried, and mounted on glass slides with Mountmedia (Wako Pure Chemical Industries, Ltd., Osaka, Japan). LM microphotographs of cleaned frustules were taken by using a Zeiss Imager Z2 microscope (Carl Zeiss Microimaging GmbH, Jena, Germany) with differential interference contrast (DIC). For scanning electron microscopy (SEM) observation, cleaned frustules were placed on round coverslips, air-dried, and coated with osmium. SEM observation was performed by using a Hitachi S–4800 (Hitachi, Ltd., Tokyo, Japan).
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