The samples were illuminated with a 100 W mercury lamp and visualized with an epifluorescence microscope (Eclipse Ti; Nikon) using an oil coupled Plan Apo 60 × 1.40 objective (Nikon). Filter blocks with UV-cut specifications (TRITC: EX540/25, DM565, BA606/55; GFP-HQ: EX455–485, DM495, BA500–545; Nikon) were used in the optical path of the microscope to allow the visualization of samples while eliminating the ultraviolet portion of the radiation and minimizing the harmful effects of ultraviolet radiation on the samples. Images and movies were captured using a cooled CMOS camera (Neo sCMOS; Andor) connected to a PC. To monitor a field of view for more than several minutes, ND filters (ND4, 25% transmittance) were inserted into the illuminating light path of the fluorescence microscope to avoid photobleaching.
Dm495
Manufactured by Nikon
The DM495 is a digital microscope camera designed for laboratory use. It features a high-resolution CMOS sensor that captures images and videos with excellent clarity and detail. The DM495 provides a convenient interface for capturing, storing, and sharing microscopic observations, enabling efficient documentation and analysis of samples.
Automatically generated - may contain errors
3 protocols using dm495
1
Fluorescence Microscopy Imaging Optimization
2
Confocal Imaging of Osteoclast Morphology
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Images of osteoclasts were acquired by confocal microscopy (A1; Nikon). The excitation laser wavelength, dichroic mirrors, and emission filters to detect EGFP were 488 nm, DM560, and BP525/50, respectively. Those for tdTomato were 561 nm, DM640, and BP595/50, respectively, and those for DAPI were 405 nm, DM495, and BP450/50, respectively (Nikon). Images of tdTomato were binarized and measured to determine the osteoclast area using NIS Elements integrated software.
3
Visualizing Microtubule Motility Dynamics
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To observe the motility of MTs, the samples were illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using an oil-coupled Plan Apo 60×1.40 objective (Nikon). UV cut-off filter blocks (GFP-HQ: EX455-485, DM495, BA500-545; Nikon) were used in the optical path of the microscope. These blocks allowed visualization of samples but eliminated the UV part of the radiation, and minimized the harmful effect of UV radiation on the samples. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. To capture a field of view for more than several minutes, an ND filter (ND4, 25% transmittance) was inserted into the illumination light path of the fluorescence microscope to avoid photobleaching of samples.
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