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Intracellular and extracellular sensing of physical and chemical variables is important for disease diagnosis and the understanding of cellular biology. Optical sensing utilizing fluorescent nanodiamonds (FNDs) is promising for probing intracellular and extracellular variables owing to their biocompatibility, photostability, and sensitivity to physicochemical quantities. Based on the potential of FNDs, we outlined the optical properties, biocompatibility, surface chemistry of FNDs and their applications in intracellular biosensing. This review also introduces biosensing platforms that combine FNDs and lab-on-a-chip approaches to control the extracellular environment and improve sample/reagent handling and sensing performance. [ABSTRACT FROM AUTHOR]

Artificial nanorobots have emerged as promising tools for a wide range of biomedical applications, including biosensing, detoxification, and drug delivery. Their unique ability to navigate confined spaces with precise control extends their operational scope to the cellular or subcellular level. By combining tailored surface functionality and propulsion mechanisms, nanorobots demonstrate rapid penetration of cell membranes and efficient internalization, enhancing intracellular delivery capabilities. Moreover, their robust motion within cells enables targeted interactions with intracellular components, such as proteins, molecules, and organelles, leading to superior performance in intracellular biosensing and organelle-targeted cargo delivery. Consequently, nanorobots hold significant potential as miniaturized surgeons capable of directly modulating cellular dynamics and combating metastasis, thereby maximizing therapeutic outcomes for precision therapy. In this review, we provide an overview of the propulsion modes of nanorobots and discuss essential factors to harness propulsive energy from the local environment or external power sources, including structure, material, and engine selection. We then discuss key advancements in nanorobot technology for various intracellular applications. Finally, we address important considerations for future nanorobot design to facilitate their translation into clinical practice and unlock their full potential in biomedical research and healthcare. [ABSTRACT FROM AUTHOR]

Intracellular and extracellular sensing of physical and chemical variables is important for disease diagnosis and the understanding of cellular biology. Optical sensing utilizing fluorescent nanodiamonds (FNDs) is promising for probing intracellular and extracellular variables owing to their biocompatibility, photostability, and sensitivity to physicochemical quantities. Based on the potential of FNDs, we outlined the optical properties, biocompatibility, surface chemistry of FNDs and their applications in intracellular biosensing. This review also introduces biosensing platforms that combine FNDs and lab-on-a-chip approaches to control the extracellular environment and improve sample/reagent handling and sensing performance.

Artificial nanorobots have emerged as promising tools for a wide range of biomedical applications, including biosensing, detoxification, and drug delivery. Their unique ability to navigate confined spaces with precise control extends their operational scope to the cellular or subcellular level. By combining tailored surface functionality and propulsion mechanisms, nanorobots demonstrate rapid penetration of cell membranes and efficient internalization, enhancing intracellular delivery capabilities. Moreover, their robust motion within cells enables targeted interactions with intracellular components, such as proteins, molecules, and organelles, leading to superior performance in intracellular biosensing and organelle-targeted cargo delivery. Consequently, nanorobots hold significant potential as miniaturized surgeons capable of directly modulating cellular dynamics and combating metastasis, thereby maximizing therapeutic outcomes for precision therapy. In this review, we provide an overview of the propulsion modes of nanorobots and discuss essential factors to harness propulsive energy from the local environment or external power sources, including structure, material, and engine selection. We then discuss key advancements in nanorobot technology for various intracellular applications. Finally, we address important considerations for future nanorobot design to facilitate their translation into clinical practice and unlock their full potential in biomedical research and healthcare.

While CRISPR/Cas12a system is a potent in vitro fluorescence biosensing toolbox, several difficulties including collective biological delivery, potential pre-activation and inadequate sensitivity make it challenging to further perform intracellular application. Herein, we raise a series of solving strategies. First, MnO 2 nanoflower is used to conjointly carry the biomolecular modules comprising CRISPR/Cas12a system through a simple physisorption to actualize an efficient endocytosis, after which the MnO 2 matrix will be reduced by the widespread intracellular biothiols to not only release the surface attached biomolecules but also produce abundant Mn2+ to enhance trans -cleavage activity. Besides, a light activation conception realized by inserting the biosensing frame with a facile photocleave group is applied to achieve a controllable spatiotemporal behavior, for which the target recognition process will be manually initiated by illuminating with an ultraviolet upconversion luminescence converted from 808 nm near-infrared photons. By making usage of hybridization chain reaction to construct a final cascaded CRISPR/Cas12a system, the additional signal amplification design renders this newly-raised CRISPR/Cas12a system-mediated fluorescent biosensor has an ultra-sensitive detection ability for microRNA-21. Moreover, this special CRISPR/Cas12a system is also equipped with a high-efficiency imaging performance in live cells and even bodies, impelling the development of CRISPR system based biosensing techniques. [Display omitted] • Near-infrared light activation conception is proposed in CRISPR/Cas12a system. • Hybridization chain reaction is used to cascade CRISPR/Cas12a system. • Mn2+ is employed to enhance the trans -cleavage activity of CRISPR/Cas12a system. • Our CRISPR/Cas12a system shows a robust intracellular biosensing ability. [ABSTRACT FROM AUTHOR]