Figure 4: GFP- c-fms monocytic lineage cells have shorter NADH lifetimes than mammary tumor cells.
From: In Vivo Visualization of Stromal Macrophages via label-free FLIM-based metabolite imaging
(A) Endogenous Fluorescence of NADHHI showing tumor cells in the breast microenvironment. (B) GFP c-fms cells in the breast tumor microenvironment. (C) SHG demonstrating collagen in the same field-of-view. Scale bar A-C = 100 μm. (D) Composite of the 3 channels shown in A-C: red = NADHHI, green = GFP-cfms, grayscale = SHG. Scale bar = 100 μm. (E) NADH-FLIM image of same field-of-view as in A-D. (F) NADH-FLIM image masked to show GFP-cfms positive cells overlayed on the NADH intensity image to provide comparison of stromal macrophages (heat map of NADH-FLIM) with NADHHI tumor cells (grayscale). (G) NADH-FLIM image masked to show NADHHI tumor cells overlayed on the SHG image to provide comparison of the tumor NADH-FLIM signature (heatmap) in the context of collagen structure (greyscale). (H) Graph showing average lifetime comparing GFP-cfms cells and tumor cells. Data points represent 4 regions of interest from 2 animals. Wilcoxon Rank sum was performed showing statistical significance with p = 0.03. (I) Graph showing the fractional contribution of the short lifetime component, free NADH, comparing GFP-cfms cells and tumor cells. Wilcoxon Rank sum was performed showing statistical significance with p = 0.03.
