In vivo enhanced bacterial
retention with bioorthogonal
mediation. (A) Fluorescence imaging of GFP and mCherry in mouse feces
after 20 h culture in LB medium. Mice in different groups were gavaged
with N3-GFP, GFP + DBCO-mCherry, N3-GFP + DBCO-mCherry. Feces of mouse
in different groups were collected 6 h after gavage (n = 3). (B) Quantitative analysis of GFP and mCherry fluorescence
intensity in mice feces of different groups. (C) Representative IVIS
images for evaluating bacterial retention in the GI tract of mice
mediated by biorthogonal reactions (n = 3). (D) Bacterial
colony-forming units of mCherry in LB agar plates for quantifying
the number of mCherry in mice colon tissues. Fluorescence imaging
of mCherry is shown on the plates. Quantitative analysis of fluorescence
intensity is also shown on the right (n = 3). (E)
Flow cytometry analysis for quantification of mCherry in mice colon
tissues. (n = 3). (F) In vitro fluorescence
imaging of mCherry in mice colon tissues after 20 h culture in LB
medium. Quantitative analysis of mCherry fluorescence intensity is
shown on the right (n = 3). (G) Quantitative analysis
of in vivo mCherry fluorescence intensity at different
time points. (H) Representative IVIS images of cultured mixture with
feces for evaluating bacterial colonization over 48 h. Feces were
collected at different time points after the gavage of mCherry (n = 3). (I) Quantification of fluorescence intensities.
Significant differences were assessed in (B), (D), (F), (G), and (I)
using one-way ANOVA (*P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; n.s., not significant). The mean values
and SEM are presented.