Extended Data Fig. 8: Photothermal imaging of entacapone accumulation by Phocaeicola dorei and faecal samples.
From: The Parkinson’s disease drug entacapone disrupts gut microbiome homeostasis via iron sequestration
a. All columns show representative images of faecal samples incubated with ENT-Hi for 6 h followed by hybridization with a fluorescently-labelled oligonucleotide probe targeting Phocaeicola dorei (Pdor) (Supplementary Table 10). Top panel: fluorescence signals from hybridized cells (FISH). Middle panel: Photothermal signal intensity from entacapone (PT) in Pdor cells only (PT signals of other cells were removed to enhance clarity). Bottom panel: PT signals overlayed with the SRS C-H signals. PT channel contrast: min 0 max 1.8. C-H channel is represented on a log scale. We reproducibly detected analogous differences when measuring at least one additional batch of samples. b. Representative images of a pure culture of Phocaeicola dorei incubated anaerobically with entacapone for 24 or 48 h. Photothermal signal from entacapone (PT) in P. dorei cells is shown in the top panel, SRS C-H signal in the middle panel, and an overlay of both signals is shown in the bottom panel. c. Single-cell specific photothermal signal intensity detected in samples shown in b. p-values were determined by the unpaired two-sided t-test comparing the groups “Ent-Hi:Fe” and “No drug”. d. Growth (assessed by microbial cell counts) of P. dorei in the presence or absence of entacapone. Data points represent the mean abundance values +/- SEM of three independent growths per condition. e. Accumulation of entacapone pre-complexed with Fe(III) by faecal samples anaerobically incubated for 6 h with ENT-Hi pre-complexed with FeSO4 (ENT-Hi:Fe). Photothermal signal from entacapone (PT) is shown on top and the overlay with the SRS C-H biomass signal is shown in the bottom panel. f. Single-cell specific photothermal signal intensity in samples shown in e. p-value was determined with the unpaired two-sided Wilcoxon test comparing the groups “Ent-Hi:Fe” and “No drug”. In c and f, boxes represent median, first, and third quartile. Whiskers extend to the highest and lowest values that are within one and a half times the interquartile range.
