Abstract
We demonstrate high-density labelling of cellular DNA and RNA using click chemistry and perform confocal and super-resolution microscopy. We visualize the crescent and ring-like structure of densely packed RNA in nucleoli. We further demonstrate click chemistry with unnatural amino acids for super-resolution imaging of outer-membrane proteins of E. coli.
Highlights
An additional request for super-resolution microscopy is a high labelling density, which is reasoned by the sampling theorem.[4]
We present a simple strategy to label the outer membrane proteins of E. coli using click chemistry in combination with an unnatural amino acid, and visualize the boundaries of bacterial cells with super-resolution imaging
We explored the speci city as well as the efficiency of the target labelling by varying the reaction times with the uorophore–azide-conjugates (Fig. S3†)
Summary
The chemical nano-environment (and the target-speci c label) of the uorophore in uences the photophysical properties[23] or may induce quenching.[24] So far, azide-substituted and red-emitting carbocyanines (e.g. Alexa Fluor 647)[18] or custom-synthesized rhodamines[19] were used for super-resolution imaging of DNA. We present click chemistry labelling of cellular nucleic acids using a variety of spectrally distinct uorophores and demonstrate confocal and single-molecule localizationbased super-resolution uorescence microscopy,[25,26,27,28] facilitating new dye combinations for multi-colour imaging.
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