Fig. 7. Comparison of virtual H& E staining using intensity, false-colour FLIM with the intensity image as the alpha channel (α-FLIM), and intensity-weighted false-colour FLIM (IW-FLIM).

(a), (h), and (o) are the true H& E staining. (e), (i), and (s) are the virtual H& E staining from intensity (b), (i), and (p), respectively. (f), (m), and (t) are the virtual H& E staining from α-FLIM (c), (j), and (q), respectively. (g), (n), and (u) are the virtual H& E staining from IW-FLIM (d), (k), and (r), respectively. The yellow and blue rectangles highlight variations in the synthetic results. The red arrows point out macrophages that are entirely absent in the intensity-based synthesis (i) but appear in the α-FLIM (j) and IW-FLIM-based (k) outcomes. Synthesis based on intensity can reconstruct macrophages when the signal is strong (bright dots pointed out by the cyan arrows in (p)), but it struggles to identify those with dimmer signals (the grey dot surrounded by the bright dots, as indicated by the cyan arrow in p). The pink arrows also indicate a few missing immune cells in intensity-based synthesis.