Embryos and larvae were imaged using a laser scanning confocal microscope Nikon A1R equipped with Nikon Plan Fluor 10x (0.30 NA) and 20x (0.50 NA) objectives. An argon ion laser (488 nm) and a diode laser (561 nm) provided the excitation light for the fluorophores. Emission light was sequentially acquired for each channel. Confocal z-stacks were processed and analyzed using Fiji software (Schindelin et al. 2012 (link)). Red channel is shown as magenta in the figures. Sections of zebrafish larvae (6 dpf) were imaged using an Epifluorescence microscope (Nikon Eclipse 90i) coupled to an Olympus DP71 digital camera.
Eclipse 90i
Manufactured by Olympus
Sourced in Japan
The Eclipse 90i is a microscope system designed for advanced optical and imaging applications. It features high-resolution optics and a modular design, allowing for customization to meet specific research needs.
Automatically generated - may contain errors
Lab products found in correlation
Plan fluor 10x, by Nikon (1 mentions)
Spe microscope, by Leica camera (1 mentions)
Dmire2 inverted microscope, by Leica camera (1 mentions)
Sp5 confocal microscope, by Leica camera (1 mentions)
Tcs sp5 confocal microscope, by Leica camera (1 mentions)
Dp70 digital camera, by Olympus (1 mentions)
Ix83 inverted microscope, by Olympus (1 mentions)
7 protocols using eclipse 90i
1
Confocal Imaging of Zebrafish Embryos
2
Fungal Morphology and Growth Characterization
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Morphological and cultural features were characterised according to Yan et al. (2015) . Briefly, mycelial discs (5 mm diam) were taken from the growing edge of 5-d-old cultures in triplicate, transferred on PDA, oatmeal agar (OA; Crous et al. 2009 ) and synthetic nutrient-poor agar medium (SNA; Nirenberg 1976 ), and incubated in the dark at 28 °C. Colony diameters were measured daily for 5 d to calculate their mycelial growth rates (mm/d). The shape, colour and density of colonies were recorded after 6 d. Moreover, the shape, colour and size of sporocarps, conidia, conidiophores, asci and ascospores were observed using light microscopy (Nikon Eclipse 90i or Olympus BX63, Japan), and 50 conidia or ascospores were measured to determine their sizes unless no or less spores were produced. Conidial appressoria were induced by dropping a conidial suspension (106 conidia/mL; 50 μL) on a concavity slide, placed inside plates containing moistened filter papers with distilled sterile water, and then incubated at 25 °C in the dark. After incubating for 24 to 48 h, the sizes of 30 conidial appressoria formed at the ends of germ tubes were measured (Yang et al. 2009 ).
3
Microscopy Imaging Techniques Protocol
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Images of transfected cells were acquired with a Zeiss Axiover 200 M inverted microscope. Nikon Eclipse 90i and Olympus BX51 microscopes were used to image histological sections. Confocal images of cells were acquired with a Leica SpE microscope (20x or 40x objective). Confocal images of microinjected or antibody-stained embryos were acquired with a Leica SP5 confocal microscope. Images were acquired with a 63× objective and 2× zoom every 2.5 μm. Images of lacZ-stained blastocysts and outgrowths were obtained with a Leica DMIRE2 inverted microscope. Images were prepared for figures using Adobe Photoshop CS5.
4
Microscopy Imaging and Video Processing
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Nikon Eclipse 90i and Olympus BX51 microscopes were used to image histological sections and a Leica TCS SP-5 confocal microscope was used for immunofluorescence imaging. Videos were exported as uncompressed files and processed in Fiji: videos were cropped to reduce file size and labels added. Processed videos were saved at the same frame rate than acquisition, 40 frames per second (fps), in JPEG compressed format.
5
Conidiophore Morphology Assessment
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Strains Hz36 and Hk37 were grown on PDA plates for two weeks in a growth chamber at 25 °C with a 12 h light/dark cycle. To assess and describe the structure and morphology of conidiophores, mycelia were taken from the edge of conidiogenous pustules or fascicles. It took 14 days for conidial induction to be observed. Microscopic morphologies such as conidia and conidiophore were observed using an optical microscope (Nikon Eclipse 90i or Olympus BX63, Tokyo, Japan).
6
Tracing Habenular Connectivity with DiI
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For each experiment, a tiny crystal of the lipophilic tracer DiI (Invitrogen) was placed on the tip of a sharp tungsten needle and inserted into the adult brain under a stereomicroscope. DiI applications to the right and left habenula (7 cases for right habenula/8 cases for left habenula) were performed approaching the brain from a dorsal aspect after removing the pineal and parapineal. Other brain structures accessed by this procedure were: olfactory bulb (2 cases), posterior area of the pallium (Dp; 2 cases), the vENT (2 cases) and the posterior hypothalamic lobe (1 case). The application area was sealed with melted 3% agarose and brains were incubated in the dark for 2–14 days in PFA at 37°C. After this period, transverse sections (50 μm thick) were cut on a vibratome, mounted on gelatin-coated slides and photographed using an epifluorescence photomicroscope (Nikon Eclipse 90i) equipped with an Olympus DP70 digital camera. Some selected sections were further imaged using a laser scanning confocal microscope (Nikon A1R).
7
Quantitative Image Analysis of TUNEL and EthD-1
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TUNEL and EthD-1 sections were analyzed using a Nikon (Mississauga, ON, Canada) Eclipse 90i digital light microscope and an Olympus (Toronto, ON, Canada) IX83 Inverted Microscope, respectively. Five images were analyzed per section for TUNEL and EthD-1 analysis and total (+) staining per field of view was quantified by ImageJ software v 1.48 (National Institutes of Health, Bethesda, MD). Background was subtracted from images and the color threshold (RGB) was adjusted uniformly to accentuate positively stained areas, which were then measured digitally.
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