Eclipse ni microscope

Soil samples were collected randomly from a pristine forest inthe Bat Xat Nature Reserve, Lao Cai Province, Vietnam. Nematodes were extracted from soil samples using modified Baermann funnel technique (Southey, 1986 ).
They were heat killed, fixed in 4% formaldehyde (for morphological observations) or in a DESS mixture (Yoder et al., 2006 (link)) (for molecular analyses), transferred to anhydrous glycerol (Seinhorst, 1962 (link)), and mounted on glass slides for microscopic observation. Measurements were performed with a Nikon digital camera on a Nikon Eclipse Ni microscope at the Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology (VAST), Vietnam. Observations of morphological diagnostic features and light microscope were taken with a Nikon digital camera mounted on a Nikon Eclipse Ni microscope. Illustrations were drawn using a Nikon Eclipse Ni microscope equipped with a Nikon Y-IDT drawing tube. Photographs and illustrations were edited by using Adobe Photoshop CC 2018.

For histology and immunostaining, 8 µm sections were cut using a microtome (SLEE medial, Mainz, Germany). Russell-Movat pentachrome (Abcam, Cambridge, UK) or Alizarin Red staining (Sigma Aldrich, St. Louis, MO, USA) were performed according to the manufacturer’s instructions. For immunohistochemistry analysis, after antigen retrieval with EDTA buffer (Sigma-Aldrich, St. Louis, MO, USA), slices were incubated overnight with an anti-BAFF-R or CD138 antibodies (Abcam, Cambridge, UK) diluted at 1:250 or 1:8,000, respectively. Slides were stained with HRP/DAB detection kit (Abcam, Cambridge, UK) and counterstained with hematoxylin to visualize cell nuclei. Images of entire aortic valve cusps were composed using NIS-Elements Microscope Imaging Software (Nikon, Tokyo, Japan) from a series of adjacent pictures taken with a 4 × objective taken with a Nikon Eclipse Ni microscope (Nikon, Tokyo, Japan). Immunostaining images were taken with a 20 × objective with a Nikon Eclipse Ni microscope (Nikon, Tokyo, Japan).

The hybridized fluorescent slides were viewed under a Nikon Eclipse Ni microscope. Images were acquired under identical conditions at ×60 magnification. Image acquisition was performed by imaging 4’,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) staining at a fixed Z Position while a Z stack of ±5 μm at 1 μm intervals was carried out. The final image was stacked to a single level before further quantification. On each sample, at least 100 cells were counted. Analysis and quantifications of these samples were performed using ImageJ software (free access). PLA dots were quantified on 8-bit images using the “Analyse Particles” command, while cells were counted using the cell counter plugin.
IHC images were also acquired using Nikon Eclipse Ni microscope at ×40 magnification and PLA dots were quantified as described above.