Humeral bone tissue of neonates or femur tissue of adult mice were dissected, fixed (4 h at 4°C for neonatal bones, and overnight at 4°C for adult bones) in 4% paraformaldehyde, frozen or embedded in paraffin, and sectioned (5-µm thickness). Adult bones were decalcified in 10% EDTA solution at 4°C for 4 wk. The tissue sections were stained with hematoxylin and eosin by using a standard procedure. Immunohistochemical and immunofluorescent staining of tissue sections was performed as described previously (Wu et al., 2015 (link)) with mouse antibodies recognizing kindlin-2 (clone 3A3.5; Tu et al., 2003 (link); Wu et al., 2015 (link)), YAP1/TAZ (sc-101199; Santa Cruz), or mouse IgG (Santa Cruz) as a control. Images were acquired at room temperature by using a microscope (Eclipse Ni; Nikon) with Plan 10×/0.25 and Plan 20×/0.4 objectives (Nikon) equipped with a digital camera (DS-Fi1c; Nikon) and NIS-Elements F Ver4.30.01 image analysis software (Nikon).
Nis elements f ver4.30.01 image analysis software
Manufactured by Nikon
NIS-Elements F Ver4.30.01 is an image analysis software developed by Nikon. It provides tools for capturing, processing, and analyzing digital images from Nikon microscopes and cameras.
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Lab products found in correlation
4 protocols using nis elements f ver4.30.01 image analysis software
1
Immunohistochemical Analysis of Bone Tissues
2
Histological Analysis of Mouse Mammary Glands
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Mouse mammary fat pads were surgically collected, spread onto a glass slide, and fixed with Carnoy’s fixative (1:3 mixture of glacial acetic acid and 100% ethanol) overnight. After hydration, mammary fat pads were stained with Carmine Alum overnight, then dehydrated and cleared in xylenes and finally mounted with Permount (Thermo Scientific, Cat# SP15-100). Stained slides were scanned with a digital camera (DS-Fi1c; Nikon) and NIS-Elements F Ver4.30.01 image analysis software (Nikon). Hyperplasia lesions in mammary glands were analyzed using Image J (NIH, USA).
3
Quantifying Lung Tumor Areas
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H&E-stained slides were scanned at × 2.5 objective magnification with a digital camera (DS-Fi1c; Nikon) and NIS-Elements F Ver4.30.01 image analysis software (Nikon). Lung tumor areas were quantified using Image J software in manual measurement mode.
4
Transwell Cell Migration Assay
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The cells (as specified in each experiment) were washed with PBS, trypsinized, and then mixed with the complete medium. The cells were pelleted by centrifugation and washed once with the medium without FBS. The cells were seeded into the transwell motility chambers with 300 μL medium without FBS (40000 cells/chamber; the pore size of the membrane inserts = 8 μm, Costar 3422), while 700 μL complete medium was added into each well of the 24-well plate in which the transwell motility chambers were placed. After incubation at 37 °C with 5% CO2 for 24 h, the cells on the upper side of the membrane inserts were removed, and the cells that had migrated to the lower side were fixed with 4% paraformaldehyde and stained with DAPI (4,6-diamidino-2-phenylindole) for 30 min at room temperature. Migrated cells in five random fields were counted with Image J, and data from three independent experiments were analyzed. The images were captured with a digital camera (DS-U3, Nikon) and NIS-Elements F Ver4.30.01 image analysis software (Nikon) with a × 20 objective.
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