What question did this study set out to answer?

The aim is to establish a dual-mode labeling strategy to track oral microbiota during intestinal translocation.

March 30, 2026Open Access

The utilization of a magneto-fluorescence sequential labeling strategy for the visualization and tracking of intestinal translocation of oral microbiota

Puntos clave

The aim is to establish a dual-mode labeling strategy to track oral microbiota during intestinal translocation.
Constructed magnetic supraparticles modified with polyacrylic acid and 3-azido-D-alanine hydrochloride (MSP@PAA-ADA)
Employed a fluorescent metabolic probe Cy5ADA for sequential labeling
Used labelling for efficient visualization of oral bacteria and monitoring their movement through the gut
Performed in vitro studies to confirm the viability and magnetic separation of labeled bacteria.
MSP@PAA-ADA showed good hydrophilicity and high magnetic responsiveness
Salivary microbiota translocated to the gut, detectable up to 36 hours post-gavage
The labeled bacteria maintained viability during tracking
Magneto-fluorescence tracking demonstrated real-time insights into the bioactivity of translocated oral bacteria.

Resumen

The microbiota of the oral-gut axis influences systemic physical health, but the exact mechanisms remain unknown. In this study, magnetic supraparticles modified with polyacrylic acid and 3-azido-D-alanine hydrochloride (MSP@PAA-ADA) were constructed, and a magnetic-fluorescence sequential labeling strategy was established for microbiota viability labeling in combination with a cyanine 5-labeled D-amino acid (Cy5ADA) fluorescent probe. MSP@PAA-ADA, with a particle size of approximately 160 nm, exhibited good hydrophilicity, high magnetic responsiveness, and low bacterial toxicity. MSP@PAA-ADA achieved efficient labeling of oral bacteria through interaction with peptidoglycan in bacterial cell walls. The experimental results indicated that MSP@PAA-ADA can be able to widely bind salivary microbiota, and the labeled salivary bacteria translocated along the oral-gut axis. A small amount of fluorescent signal could still be detected in the mice 36 hours after gavage. MSP@PAA-ADA was employed as a magnetic probe for the in vitro labeling of oral bacteria, and sequential labeling was conducted using the fluorescent metabolic probe Cy5ADA. The oral bacteria labeled with MSP@PAA-ADA can be effectively magnetically separated after intestinal translocation through the gastrointestinal tract. The magnetically responsive and fluorescently imaged oral bacteria demonstrate their activity after intestinal translocation. The magneto-fluorescence dual-mode strategy that enables controllable spatiotemporal tracing and simultaneous viability assessment of intestinal oral bacteria in the gut, providing a new technical approach for dissecting biological mechanisms along the oral–gut microbiota axis. • MSP@PAA-ADA enables efficient labeling of salivary bacteria without affecting viability. • Co-labeling of MSP@PAA-ADA and Cy5ADA reveals salivary microbiota intestinal translocation. • Magneto-fluorescence tracking reveals transient bioactivity of oral bacteria in gut. • The strategy enables real-time longitudinal tracking for oral-gut axis microbiota research.

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