Journal: Nature Communications

Article Title: FRET imaging of glycoRNA on small extracellular vesicles enabling sensitive cancer diagnostics

doi: 10.1038/s41467-025-58490-2

Figure Lengend Snippet: a Schematic illustration of in situ FRET imaging of glycoRNAs. ISHPs were used for in situ hybridization of RNAs, whereas GRPs for specific targeting Neu5Ac. FRET occurs only when dual recognition of Neu5Ac and RNAs within a proximity of 10 nm. b Schematic of the confocal FRET imaging of sEV glycoRNAs. The bead-bound sEVs were uniformly deposited onto the focal plane of the microscope, and the FRET signal was automatically corrected using the sensitized emission method. The FRET efficiency and distance between GRPs (donor) and ISHPs (acceptor) were calculated. c Fluorophore photophysical properties, and our selection of wavelength for excitation (559 nm, green), optical filter (585 nm), and FRET collection band (650–700 nm, cyan). Excitation/readout optics and calculated inter-dye Förster radii ( R da ). The ultraviolet-visible diffuse reflectance spectrum of the optically inert carrier fcPS is represented by the gray dashed line, which exhibits no absorption within the drFRET optical analysis bands.

Article Snippet: The NC membrane was subsequently placed in a hybridization oven (LF-III, Ningbo Scientz Biotechnology Co., Ltd., China) and washed twice with 2 × SSC (0.1% SDS, 5 min, 30 °C, 414 × g ) and twice more with 0.5 × SSC (0.1% SDS, 15 min, 30 °C, 414 × g ).

Techniques: In Situ, Imaging, In Situ Hybridization, Microscopy, Selection