Mice were perfused by cardiac puncture with PBS containing 10U ml−1 of heparin and then 4% PFA, followed by overnight post-fix with 10% formalin. For en face staining, the adventitia was thoroughly cleaned under dissecting microscope, and the aorta was cut open longitudinally and pinned on parafilm. Plaques were stained with Oil red O and photographed with a digital camera. For cross-sectional quantification of plaque progression, 5μm serial paraffin sections from the indicated sites of the aorta were stained with hematoxylin and eosin. Plaques were imaged with a Nikon Eclipse 80i microscope connected to a digital camera. Plaque area was then quantified with NIS.Elements D3.2 imaging software (Nikon Inc.) to obtain the maximum cross-sectional plaque area for each site. Fibrous cap thickness was quantified by choosing the largest necrotic core in a section and measuring the thinnest part of the fibrous cap. Representative images were finally processed with Photoshop (Adobe).
Nis elements d3.2 imaging software
Manufactured by Nikon
NIS-Elements D3.2 is an imaging software designed for microscopy applications. It provides a comprehensive platform for image acquisition, processing, and analysis. The software supports a wide range of imaging techniques, including fluorescence, brightfield, and confocal microscopy. NIS-Elements D3.2 offers tools for image stitching, deconvolution, and advanced image analysis, catering to the needs of researchers and scientists in various fields.
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3 protocols using nis elements d3.2 imaging software
1
Quantitative Analysis of Atherosclerotic Plaques
2
Quantitative Analysis of Atherosclerotic Plaques
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Mice were perfused by cardiac puncture with PBS containing 10U ml−1 of heparin and then 4% PFA, followed by overnight post-fix with 10% formalin. For en face staining, the adventitia was thoroughly cleaned under dissecting microscope, and the aorta was cut open longitudinally and pinned on parafilm. Plaques were stained with Oil red O and photographed with a digital camera. For cross-sectional quantification of plaque progression, 5μm serial paraffin sections from the indicated sites of the aorta were stained with hematoxylin and eosin. Plaques were imaged with a Nikon Eclipse 80i microscope connected to a digital camera. Plaque area was then quantified with NIS.Elements D3.2 imaging software (Nikon Inc.) to obtain the maximum cross-sectional plaque area for each site. Fibrous cap thickness was quantified by choosing the largest necrotic core in a section and measuring the thinnest part of the fibrous cap. Representative images were finally processed with Photoshop (Adobe).
3
Microscopic Imaging of Lipid Staining
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A Nikon Microphot FXA microscope equipped with a DS-Fi1 camera and the NIS Elements D.32 imaging software (Nikon instruments Europe B.V., Badhoevedorp, The Netherlands) and a Leica multispectral confocal laser microscope (Leica TCS NT, Heidelberg, Germany) were used to examine the cryosections. Oil red O-stained lipids were detected under the confocal microscope using an excitation laser line at 543 nm (helium-neon). Optical sections (1.0 µm) were acquired, and images were produced using an 8-fold frame averaging a 1024 × 1024 pixel resolution. Representative images were presented using Adobe Creative Cloud (Adobe Inc., San Jose, CA, USA).
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