Abstract
Optical super-resolution techniques reach unprecedented spatial resolution down to a few nanometers. However, efficient multiplexing strategies for the simultaneous detection of hundreds of molecular species are still elusive. Here, we introduce an entirely new approach to multiplexed super-resolution microscopy by designing the blinking behavior of targets with engineered binding frequency and duration in DNA-PAINT. We assay this kinetic barcoding approach in silico and in vitro using DNA origami structures, show the applicability for multiplexed RNA and protein detection in cells, and finally experimentally demonstrate 124-plex super-resolution imaging within minutes.
🔬 Techniques
✨ Fluorophores
🧪 Sample Preparation
🔬 Cell Lines
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1
Simultaneousnmultiplexed super-resolution imaging by engineeringnblinking kinetics. (a) Engineering blinking kinetics in DNA-PAINTnallows the creation of u201cbarcodesu201d for simultaneous multiplexi...
Figure 2
Engineerednbinding kinetics allow simultaneous multiplexed super-resolutionnimaging of RNA and proteins in cells. (a) Scheme showing the implementationnof frequency barcoding for smRNA-FISH. Two disti...
Figure 3
Frequency-based 124-plexnsuper-resolution imaging. (a) DNA origaminstructures are extended with three unique sequences (red, green, ornblue) with 0, 3, 9, 22, or 44 copies, respectively. Using combina...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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