Endogenous NO levels were estimated by using the NO-sensitive dye DAF-FM DA (Kojima et al., 1999 (link); Moreau et al., 2008 (link)). Roots were observed by fluorescence microscopy and bright-field microscopy using an Eclipse E200 microscopy (Nikon). After Kan selection, seedlings were transferred for 7 days to plates containing agar and modified Hoagland solution as described above. For nitrate and protein determination, 100 mg of seedlings were ground in liquid N2 and resuspended in 100 mM sodium phosphate pH 7.4. After centrifugation at 10,000 g for 15 min at 4°C, supernatants were used for nitrate determination as described by Cataldo et al. (1975) (link). Samples were incubated with Salicylic acid (50 mg/ml) and reaction was stopped with 2N NaOH. Absorbance was measured at 410 nm. Protein content were determined by the Bradford method (Bradford, 1976 (link)).
Eclipse e200 microscopy
Manufactured by Nikon
The Eclipse E200 is a laboratory microscope designed for various applications. It features a sturdy and stable construction with a built-in illumination system. The microscope provides clear and detailed images through its optical components.
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Lab products found in correlation
4 protocols using eclipse e200 microscopy
1
Quantifying Endogenous Nitric Oxide Levels
2
Histological Evaluation of EAE-Induced Mice
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Mice induced with EAE at 18th d to 21st d were sacrificed. Spinal cords were harvested and fixed in 10% neutral-buffered formalin and processed routinely for paraffin embedment. Slides were stained with hematoxylin-and-eosin and the images were captured by Nikon Eclipse E200 microscopy (Melville, NY). Histological lesion was evaluated by counting the inflammatory foci (>10 mononuclear cells) in the meninges and parenchyma in a blinded fashion in that the pathologist was unaware of the clinical status.
3
Boyden Chamber Assay for Cell Migration
4
Boyden Chamber Assay for Cell Migration
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A Boyden chamber assay was used, as described previously (17, 29 (link)). Briefly, cells were trypsinized, suspended in 0.1% BSA/RPMI, and seeded (2.5 × 104 cells/well) in the upper compartment of a Boyden chamber (NeuroProbe). A 12-μm-pore Matrigel-coated polycarbonate membrane was used to separate the upper and lower compartments. In the lower chamber, RPMI medium containing 10% FBS was used. After an incubation period of 16 hours at 37°C, membranes were recovered and cells on the upper side of the membrane (nonmigrating) were wiped off the surface. Migrating cells on the lower side of the membrane were fixed and stained with the Hema 3 Staining kit (Thermo Fisher Scientific). Migrating cells in each well were counted in five random fields by contrast microscopy using an Eclipse E200 Nikon microscopy (4X magnification) and the ImageJ/Fiji software.
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