Fluorescent carbon dots for labeling of bacteria: mechanism and prospects-a review.

The search for bacteria-labeling agents that are more efficient and less toxic compared to existing staining dyes is ongoing. Fluorescent quantum dots and carbon dots (CDs) have been extensively researched for various bioimaging applications. Priority is given to CDs due to several advantages, including lower toxicity, versatility in tuning their properties, and better photostability compared to metal-based quantum dots. Although significant progress is still needed to replace existing dyes with CDs for bacteria labeling, they offer promising potential for further improvement in efficiency. Surface charges and functional groups have been reported as decisive factors for bacterial discrimination and live/dead assays; however, a complete guideline for preparing CDs with optimum properties for efficient staining and predicting their labeling performance is lacking. In this review, we discuss the application of fluorescent CDs for bacterial labeling and the underlying mechanisms and principles. We primarily focus on the application and mechanism of CDs for Gram differentiation, live imaging, live/dead bacteria differentiation, bacterial viability testing, biofilm imaging, and the challenges associated with application of CDs. Based on proposed mechanisms of bacterial labeling and ambiguous results reported, we provide our view and guidelines for the researchers in this field to overcome the challenges associated with bacteria labeling using fluorescent CDs.