Practical fluorescence reconstruction microscopy for large samples and low-magnification imaging
Fig 2
Low-magnification nuclei reconstruction.
(A) Representative transmitted-light image of MDCK cells at 5x magnification, with corresponding: (B) ground-truth nuclei, stained with Hoecscht 33342 and imaged with blue fluorescent light; (C) nuclear prediction produced by the network; and (D) the overlay of (B) and (C) displayed in red and green, respectively. The raw accuracy score between (B) and (C) is given at right. The scale bar is 100 μm. (E) Representative transmitted-light image of keratinocyte cells at 10x magnification, with corresponding (F, G, H) ground truth nuclei image, predicted nuclei, and overlay, respectively. Scale bar is 50 μm. (I) Comparison of the accuracy score distributions across the 5X MDCK and 10X Keratinocyte datasets, N > 4400 test images for each dataset (see S1 Table). (J,K) A comparison of nuclear area estimations and centroid-centroid displacement estimations, respectively, for the two low-magnification datasets considered here. See Methods. (L) Pairwise comparisons of predicted cell counts for MDCK (left) and Keratinocytes (right). Summary statistics and N shown below plots. (M-N) Sequence of phase images (M-M”) from a time-lapse at 0, 12, and 24 hours of growth, with corresponding nuclear predictions (N-N”) respectively. Input data consists of MDCK WT cells imaged at 5x magnification and montaged; the U-Net was applied in a sliding-window fashion to predict small patches of the image in parallel. Scale bar is 1 mm. Higher resolution movies showing the migration dynamics can be seen in S1 Movie.