Fig. 3: Photothermal imaging of entacapone bioaccumulation by microbiota cells.
From: The Parkinson’s disease drug entacapone disrupts gut microbiome homeostasis via iron sequestration
a, Schematic illustration of the time-dependent signal obtained from a solution of 10 mM entacapone in DMSO, with photothermal (w/ PT) and without photothermal (w/o PT) detection. b, Photothermal signal intensity distribution from entacapone (PT), and SRS C-H signal of live and dead (PFA fixed) microbiota cells incubated in the presence (+) or absence (−) of ENT-Hi for 6 h. Scale bar, 10 μm. c, Single-cell photothermal signal intensity distribution in samples shown in b. P values were determined using unpaired two-sided Wilcoxon test comparing groups ENT-Hi and No drug, or ENT-Hi and Dead+ENT-Hi (Supplementary Table 12). d, Correlation between activity (%CDSRS) and PT signal intensity for microbiota cells exposed to ENT-HI for 6 h. Pearson correlation coefficient (R) and two-sided P value are indicated. Incubations were established using a new mixture of samples from two donors in sM9. e, Representative images of the original mixture of 6 faecal samples incubated with ENT-Hi for 6 h followed by hybridization with FISH probes targeting E. coli (Ecol), Streptococcus and Lactococcus (Strc), Ruminococcus bromii (Rumi), B. uniformis (Buni) and P. dorei (Pdor) (Supplementary Table 10). Top: FISH-probe signals from hybridized cells. Middle: PT signal maps from entacapone (cells targeted by FISH are shown with white contour lines). Bottom: entacapone PT signals overlayed with SRS C-H signals. Scale bar, 8 μm. In b and e, PT channel contrast is min 0 and max 1.8, and C-H signals are represented on a log scale. f, Single-cell photothermal signal distribution in samples shown in c. P values were determined using unpaired two-sided Wilcoxon test comparing ENT-Hi and No drug groups of targeted taxa only. g, Absolute abundance of taxa targeted by FISH in No drug, Ent-Low and ENT-Hi incubations (Supplementary Table 4). Data represent the mean ± s.d. of 3 incubation vials. In c and f, boxes represent the median, first and third quartile. Whiskers extend to the highest and lowest values that are within 1.5× the interquartile range.
