Fluorescence can be enhanced or quenched depending on the distance between the surface of a metal nanoparticle and the fluorophore molecule. Fluorescence enhancement by nearby metal particles is called metal-enhanced fluorescence (MEF). MEF shows promising potential in the field of fluorescence-based biological sensing. MEF-based biosensor systems generally fall into two platform categories: (1) a two/three-dimensional scaffold, or (2) a colloidal suspension. This review briefly summarizes the application studies using wavelength-dependent carbon dots (UV-VIS), noble metals (VIS), and upconversion nanoparticles (NIR to VIS), representative nanomaterials that contribute to the enhancement of fluorescence through the resonance energy transfer modulation and then presents a perspective on this topic.

The integration of deep learning (DL) and nanozyme-based biosensing has emerged as a transformative strategy for next-generation medical diagnostics. This review explores how DL architectures enhance nanozyme design, functional optimization, and predictive modeling by elucidating catalytic mechanisms such as dual-atom active sites and substrate-surface interactions. Key applications include disease biomarker detection, medical imaging enhancement, and point-of-care diagnostics aligned with the ASSURED criteria. In clinical contexts, advances such as wearable biosensors and smart diagnostic platforms leverage DL for real-time signal processing, pattern recognition, and adaptive decision-making. Despite significant progress, challenges remain—particularly the need for standardized biomedical datasets, improved model robustness across diverse populations, and the clinical translation of artificial intelligence (AI)-enhanced nanozyme systems. Future directions include integration with the Internet of Medical Things, personalized medicine frameworks, and sustainable sensor development. The convergence of nanozymes and DL offers unprecedented opportunities to advance intelligent biosensing and reshape precision diagnostics in healthcare.

Recent advances in nanotechnology and optical imaging have transformed molecular diagnostics, enabling the detection and analysis of individual biomolecules with unprecedented precision. Nanobiosensors provide ultrasensitive molecular detection, and super-resolution microscopy (SRM) exceeds the diffraction limit of conventional optics to achieve nanometer-scale resolution. Although their integration remains in its infancy, with only a handful of proof-of-concept studies reported, the convergence of nanobiosensors and SRM holds significant promise for next-generation diagnostics. In this review, we first outline nanobiosensor-based single-molecule detection strategies and highlight representative implementations. These include plasmonic–SRM hybrids, electrochemical–optical correlatives, and SRM-enabled immunoassays, with a focus on their applications in oncology, infectious diseases, and neurodegenerative disorders. Then, we discuss emerging studies at the interface of nanobiosensors and SRM, including nanostructure-assisted SRM. Despite not being true biosensing approaches, these studies provide valuable insights into how engineered nanomaterials can improve imaging performance. Finally, we evaluate current challenges, including reproducibility, multiplexing, and clinical translation, and outline future opportunities, such as the development of photostable probes, artificial intelligence-assisted image reconstruction, microfluidic integration, and regulatory strategies. This review highlights the synergistic potential of nanobiosensors and SRM, outlining a roadmap toward clinically translatable next-generation single-molecule diagnostic platforms.

Exosomes are nanoscale extracellular vesicles (EVs) that carry biomolecular signatures reflective of their parent cells, making them powerful tools for non-invasive diagnostics and therapeutic monitoring. Despite their potential, clinical application is hindered by challenges such as low abundance, heterogeneity, and the complexity of biological samples. To address these limitations, plasmonic biosensing technologies—particularly propagating surface plasmon resonance (PSPR), localized surface plasmon resonance (LSPR), and surface-enhanced Raman scattering (SERS)—have been developed to enable label-free, highly sensitive, and multiplexed detection at the single-vesicle level. This review outlines recent advancements in nanoplasmonic platforms for exosome detection and profiling, emphasizing innovations in nanostructure engineering, microfluidic integration, and signal enhancement. Representative applications in oncology, neurology, and immunology are discussed, along with the increasingly critical role of artificial intelligence (AI) in spectral interpretation and diagnostic classification. Key technical and translational challenges—such as assay standardization, substrate reproducibility, and clinical validation—are also addressed. Overall, this review highlights the synergy between exosome biology and plasmonic nanotechnology, offering a path toward real-time, precision diagnostics via sub-femtomolar detection of exosomal miRNAs through next-generation biosensing strategies.