Fluorescent images of stained cells were acquired using a Nikon TE300 epifluorescence microscope (Nikon, Melville, NY). A 9-by-9 scanning grid comprising of 81 image fields (with a ∼ 10% overlap with adjacent fields to reduce artificial fragmentation of cell and nucleus) were generated using the NIS Elements software. The size of each image field (based on the Nikon QMiMc camera sensor) was 1280 × 1024 pixels, and the pixel size using the 10× objective was 0.57 μm, respectively. Two channels (UV, RFP) were acquired for each image field, followed by a calibration image of both channels respectively for use in computationally reducing the non-uniformity of the illumination field (Chen et al., 2013 (link)).
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Fluorescent Microscopy Imaging of Stained Cells
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Corresponding Organization : Johns Hopkins University
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Variable analysis
independent variables
- Fluorescent images of stained cells
- Measurements of cell and nucleus
- Nikon TE300 epifluorescence microscope (Nikon, Melville, NY)
- 9-by-9 scanning grid comprising of 81 image fields
- Size of each image field (1280 × 1024 pixels)
- Pixel size using the 10× objective (0.57 μm)
- Two channels (UV, RFP) acquired for each image field
- Calibration image of both channels for computational reduction of non-uniformity in the illumination field
- Not explicitly mentioned
- Not explicitly mentioned
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