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. 2021 Mar 18;1:627626. doi: 10.3389/fbinf.2021.627626

FIGURE 2.

FIGURE 2

Acquiring mitotic cells at high resolution with AutoScanJ (confocal fluorescence microscopy). A to F Image analysis workflow with a cropped region from the primary scan map and a single mitotic cell closer view (inset). (A) Raw image, (B) Median filter, (C) ImageJ Remove outliers, (D) Subtracting image C to image B, (E) Thresholding non-zero pixels, (F) Connected particles of area greater or equal than 12 pixels (in red) overlaid over image A. (G) Mitotic cells detected during primary scan (yellow crosses from ImageJ, enhanced in inset). (H) Zoomed area with four mitotic cells (colour boxes).(I) Interactive montage of all detected mitotic cells, including corresponding colour boxes of H]. (J) Zoomed image of a detected mitotic cell (primary scan) and (K) The same cell acquired during the secondary scan, DNA (green), tubulin (red) and centrosomes (blue). All the images shown are z-stacks maximum intensity projections. Scale bars: A–F, H - 50 µm, G - 500 µm, H - 50 µm, K - 5 µm.