Figures and data
FLIM of NAD(P)H in monolayer cell cultures.
(A) Representative FLIM images of colorectal cancer cell lines. Scale bar = 50 μm. For FLIM: ex. 750 nm, reg. 450–490 nm. (B) The relative contribution of free NAD(P)H (a1, %) in cell cultures. Box shows the median and the quartiles Q1 and Q3, whiskers show minimum and maximum. Dots indicate individual cells (n=280 for HT29 cells, n=185 for HCT116 cells, n=30 for CaCo2 cells, n=138 for CT26 cells). (C) The distribution of the NAD(P)H-a1 for the cell lines. The bimodality index (BI-a1) is shown on each diagram.
The Bimodality Index BI-a1 and Dispersity D-a1 of NAD(P)H in cultured cells, mouse tumors and patients’ tumor samples.
FLIM of NAD(P)H in mouse tumors in vivo.
(A) FLIM images of NAD(P)H of tumor cells in mouse models in vivo. Scale bar = 50 μm. For FLIM: ex. 750 nm, reg. 450–490 nm. (B) Representative histological slices of tumors, hematoxylin/eosin (HE) staining, initial magnification 20×. Scale bar = 50 μm. (C) The relative contribution of free NAD(P)H (a1, %) in tumors (t1, t2, t3) obtained from different cell lines. Box shows the median and the quartiles Q1 and Q3, whiskers show minimum and maximum. Dots are the measurements from the individual cells. The bimodality index (BI-a1) is shown above each box. (D) Representative distributions of the NAD(P)H-a1 for each type of tumor. The bimodality index (BI-a1) is shown on the diagrams. Dots indicate individual cells (n=280 for HT29, n=340 for HCT116, n=160 for CaCo2, n=350 for CT26).
FLIM of NAD(P)H in patients’ tumor samples ex vivo.
(A) Representative FLIM images of tumors. Scale bar = 50 μm. For FLIM: ex. 750 nm, reg. 450–490 nm. (B) Histopathology of tumors, hematoxylin/eosin (HE) staining, initial magnification 20×. Scale bar = 50 μm. (C) The relative contribution of free NAD(P)H (a1, %) in patients’ tumors (p1–p21). Box shows the median and the quartiles Q1 and Q3, whiskers show minimum and maximum. Dots are the measurements from the individual cells. The bimodality index (BI-a1) is shown above each box. (D) Representative distributions of the NAD(P)H-a1 for patients’ tumors.
The relationships between fluorescence decay parameters of NAD(P)H and clinicopathological characteristics of patients’ tumors.
(A) Plots of SHAP analysis for the built decision tree models to determine the importance of the parameters. The higher the parameter value, the more red the dot is. (B) Box-plots of the selected parameters with highest significance, * p-val < 0.05.
Information about patients and their colorectal tumors.
FLIM of NAD(P)H in monolayer cell cultures.
The distribution of the NAD(P)H-τm for the cell lines. The bimodality index (BI-τm) is shown on each diagram.
Immunohistochemical analysis of НСT116 and HT29 tumors using primary antibodies to EpCAM (green, epithelial cells marker) and vimentin (red, mesenchymal cells marker).
(A) Immunofluorescence images of HT29 and HCT116 tumors. Scale bar = 100 μm. (B) The ratio of positively stained EpCAM+ and vimentin+ cells in tumor sections.
The relationships between parameter BI τm and clinicopathological characteristics of patients’ tumors.
Box-plots of the clinicopathological characteristics and parameter BI-τm. No significant differences were found between the groups.
NAD(P)H fluorescence decay parameters of colorectal cancer cells in monolayer cultures in vitro and in mouse tumors in vivo.
τm – mean lifetime, τ1 – short lifetime component, τ2 – long lifetime component, a1 – relative contribution of the short lifetime component, BI-τm – bimodality index of the mean lifetime, BI-a1 – bimodality index of relative contribution of the short lifetime component, D-a1 – dispersion of relative contribution of the short lifetime component.
Clinicopathological characteristics of patients’ tumors
