Fluorescent images were obtained using a Nikon 50i epifluorescence microscope equipped with X-Cite series 120 illuminator (EXFO Photonics Solutions Inc.) and appropriate fluorescence excitation/emission filters. Grayscale images were captured using Nikon NIS-Elements software and an attached Photometrics CoolsnapEZ digital camera, pseudo-colored and merged. Alternatively, fluorescent images were obtained using a TissueFAXS Plus (Tissue Gnostics) automated microscopy workstation with a Pixelfly USB monochrome camera (PCO). Fluorescent background was subtracted using Adobe Photoshop software. Brightfield images were obtained using a Nikon 50i epifluorescence microscope, Nikon NIS-Elements software, and an attached Nikon Digital Sight DS-Fi1 camera.
50i epifluorescence microscope
Manufactured by Nikon
Sourced in Japan
The Nikon 50i epifluorescence microscope is a specialized laboratory equipment designed for conducting fluorescence-based microscopy. It provides a core function of illuminating and detecting fluorescent signals within specimens, allowing researchers to visualize and analyze various biological structures and processes.
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Lab products found in correlation
Digital sight ds fi1 camera, by Nikon (1 mentions)
Nis elements software, by Nikon (1 mentions)
Tissuefaxs plus, by TissueGnostics (1 mentions)
Pluronic acid, by Thermo Fisher Scientific (1 mentions)
Moticam pro 282b camera, by Motic (1 mentions)
Isopore polycarbonate filter, by Merck Group (1 mentions)
3 protocols using 50i epifluorescence microscope
1
Fluorescence Microscopy Imaging Protocol
2
Cytosolic Calcium Imaging in Wound Healing
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Modifications in cytosolic calcium were assessed by Fluo-4 fluorescence (Molecular Probes Inc., Eugene, OR). This probe was kept as a 1.7 mM stock solution in DMSO at -20°C and, for the experiments, mixed with 20% pluronic acid (Molecular Probes Inc.) in DMSO, and diluted to a final concentration of 1.7 μM in the corresponding solution. The cells grown on coverslips were incubated in this solution at room temperature (RT) for 15 min. After washing with CS, the coverslips were mounted on a homemade chamber [53 (link)] containing CS and 1 μg/ml propidium iodide to reveal dead cells. Wounds were produced under the fluorescent microscope and photographs taken every 1 second employing a Moticam Pro 282B camera and the Moticam Image Advanced 3.2 software. For the studies we employed a direct Nikon 50i epifluorescence microscope (Nikon, Tokyo, Japan) with a 10X Plan-Fluor objective and a B-2A Blue Excitation Filter Block (450-490 nm Excitation Filter, 500 nm Dichromatic Mirror Cut and 515 nm Barrier Filter).
3
Quantifying Sediment Bacterial Abundance
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Sediment cores were sectioned from surface to 2 cm. Sub-samples of ≈ 5 g wet weight sediment were extruded using syringe cores. The sub-samples were transferred to 45 mL of filter sterilized full strength seawater (10−1 dilution). Tween 80 (50 μL) was added and the mixture was sonicated at 40 mHz for 15 s. The sub-samples were diluted with filter sterilized seawater and fixed with buffered formalin (2% final concentration).
Enumeration of total bacteria was carried out by epifluorescence microscopy (Hobbie et al., 1977 ). A sub-sample was stained with acridine orange (final concentration 0.01% w/v), incubated for 2 min and then filtered through 0.2 μm black Isopore polycarbonate filters (Millipore). Bacterial cells were counted using a Nikon 50i epifluorescence microscope equipped with a 100X oil immersion objective. Cells were counted from 10–100 microscopic fields. Total bacterial counts were expressed as cells g−1 (dry sediment).
Enumeration of total bacteria was carried out by epifluorescence microscopy (Hobbie et al., 1977 ). A sub-sample was stained with acridine orange (final concentration 0.01% w/v), incubated for 2 min and then filtered through 0.2 μm black Isopore polycarbonate filters (Millipore). Bacterial cells were counted using a Nikon 50i epifluorescence microscope equipped with a 100X oil immersion objective. Cells were counted from 10–100 microscopic fields. Total bacterial counts were expressed as cells g−1 (dry sediment).
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