Fig. 7

FGF2-differentiated live organoids with distinct Raman signature. A Average Raman spectra for cell dense regions of control, EGF-treated, and FGF2-treated organoids, with error bars representing the 95% confidence interval. B Representative images of salivary gland organoids that were first subjected to live Raman imaging prior to fixation and immunostaining to confirm the proacinar (AQP5) cells in FGF2-treated organoids, additional stains for epithelial cells (EpCAM), ductal cells (K7), and nuclei (DAPI). Scale bar, 100 µm. Larger images and quantification shown in Additional file 1: Fig. S5. C Classification accuracy of machine learning (RSVM) on live Raman organoid dataset (228–267 spectra per comparison, Standard deviation less than 0.10 for all comparisons). D–L Multivariate SVD analysis of representative experiment with control versus FGF2 (D–F), control versus EGF (G–I), and EGF versus FGF2 (J–L). For each comparison, there is an SVD plot where each dot represents a single spectrum, with clustering of similar spectra (D, G, J). The distribution of the spectra in each region is plotted in E, H, and K, while the SVD components producing the distribution are plotted in F, I, and L. Select regions of interest within the component graphs are highlighted in yellow (n = 40+ separate spectra collected from a minimum of three independent organoids per condition, representative experiment shown of three replicate experiments)