Carbon Dot Blinking Enables Accurate Molecular Counting at Nanoscale Resolution
Open Access
- 18 February 2021
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 93 (8), 3968-3975
- https://doi.org/10.1021/acs.analchem.0c04885
Abstract
Accurate counting of single molecules at nanoscale resolution is essential for the study of molecular interactions and distribution in subcellular fractions. By using small-sized carbon dots (CDs), we have now developed a quantitative single-molecule localization microscopy technique (qSMLM) based on spontaneous blinking to count single molecules with a localization precision of 10 nm, which can be accomplished on conventional microscopes without sophisticated laser control. We explore and adapt the blinking of CDs with diverse structures and demonstrate a counting accuracy of >97% at a molecular density of 500 per μm2. When applied to G-protein coupled receptors on a cell membrane, we discriminated receptor oligomerization and clustering and revealed ligand-regulated receptor distribution patterns. This is the first example of adapting nanoparticle self-blinking for molecular counting, and this demonstrates the power of CDs as SMLM probes to reliably decipher sub-diffraction structures that mediate crucial biological functions.Keywords
Funding Information
- Ministry of Education of the People's Republic of China (18CX02126A)
- National Natural Science Foundation of China (21874154)
This publication has 58 references indexed in Scilit:
- Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM)Proceedings of the National Academy of Sciences, 2012
- The lipid raft hypothesis revisited – New insights on raft composition and function from super‐resolution fluorescence microscopyBioEssays, 2012
- Evaluation of fluorophores for optimal performance in localization-based super-resolution imagingNature Methods, 2011
- Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysisNature Methods, 2011
- Quantitative Photo Activated Localization Microscopy: Unraveling the Effects of PhotoblinkingPLOS ONE, 2011
- Identification of clustering artifacts in photoactivated localization microscopyNature Methods, 2011
- Optimized localization analysis for single-molecule tracking and super-resolution microscopyNature Methods, 2010
- Single Nonblinking CdTe Quantum Dots Synthesized in Aqueous Thiopropionic AcidAngewandte Chemie-International Edition, 2006
- Imaging Intracellular Fluorescent Proteins at Nanometer ResolutionScience, 2006
- Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)Nature Methods, 2006