Abstract
First reported in 1994, stimulated emission depletion (STED) microscopy has long been regarded as a powerful tool for real-time superresolved bioimaging . However, high STED light power (101∼3 MW/cm2) is often required to achieve significant resolution improvement, which inevitably introduces phototoxicity and severe photobleaching, damaging the imaging quality, especially for long-term cases. Recently, the employment of nanoprobes (quantum dots, upconversion nanoparticles, carbon dots, polymer dots, AIE dots, etc.) in STED imaging has brought opportunities to overcoming such long-existing issues. These nanomaterials designed for STED imaging show not only lower STED power requirements but also more efficient photoluminescence (PL) and enhanced photostability than organic molecular probes. Herein, we review the recent progress in the development of nanoprobes for STED imaging, to highlight their potential in improving the long-term imaging quality of STED microscopy and broadening its application scope. We also discuss the pros and cons for specific classes of nanoprobes for STED bioimaging in detail to provide practical references for biological researchers seeking suitable imaging kits, promoting the development of relative research field.
Highlights
Photoluminescence (PL) microscopy imaging has long been a powerful tool in biological research
Developing nanoprobes for stimulated emission depletion (STED) imaging provides a valuable view on improving the STED imaging quality from a material perspective
A variety of nanoprobes have demonstrated their applicability in realistic bioimaging of subdiffraction biostructures, both in fixed and living cells
Summary
Photoluminescence (PL) microscopy imaging has long been a powerful tool in biological research. QDots feature higher brightness and photostability, better monochromaticity, and continuously tunable emission (determined by particle size), which altogether facilitate their applications as imaging probes Despite their composition with heavy metal elements (Cd, Pb, etc.), the toxicity of QDots is still proved acceptable for in vitro research (Gao et al, 2005). The STED resolution of nucleus structure and tunneling nanotubes of CDot-stained 4T1 cells was 19.7 and 75 nm, respectively (Figure 8), and considerably improved compared with confocal imaging results under low STED power (39.6 mW). Further improvement in resolution is required to promote realistic applications of NP-STED in bioimaging To this end, Hell and Sivan et al further developed 50 nm LPR hybrid particles with gold core and a silica shell doped with molecular dye Atto488.
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