ArticleProceedings of the National Academy of Sciences of the United States of America2018
Excitation-multiplexed multicolor superresolution imaging with fm-STORM and fm-DNA-PAINT.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- Advancement of Peptide-PAINT: Applications and Current Challenges in Super-Resolution Imaging.Small science · 2026Review
- Brightness demixing for simultaneous multi-target imaging in 3D single-molecule localization microscopy.Nature methods · 2026Article
- Chromenylium and Flavylium Polymethine Fluorophores Light Up the Shortwave Infrared Region.Accounts of chemical research · 2026Article
- Machine Learning in Single-Molecule Tracking Analysis of Superresolution Optical Microscopy Data.Cells · 2026Review
- Discriminating single-molecule binding events from diffraction-limited fluorescence.Nature communications · 2025Article
- DNA Origami Multicolor Quantum Dot Platforms for Sub-diffraction Spectral Separation Imaging.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Simultaneous multicolor fluorescence imaging using PSF splitting.Nature methods · 2024Article
- Sorting of secretory proteins at theeLife · 2024Article
- Blinking characteristics of organic fluorophores for blink-based multiplexing.Communications chemistry · 2024Article
- Imaging Protein Aggregates in Parkinson's Disease Serum Using Aptamer-Assisted Single-Molecule Pull-Down.Analytical chemistry · 2023Article
- Tetra-color superresolution microscopy based on excitation spectral demixing.Light, science & applications · 2023Article
- Multiplexed Detection of Molecular Interactions with DNA Origami Engineered Cells in 3D Collagen Matrices.ACS applied materials & interfaces · 2022Article
- MiOS, an integrated imaging and computational strategy to model gene folding with nucleosome resolution.Nature structural & molecular biology · 2022Article
- Engineering a DNA origami mediated multicolour quantum dot platform for sub-diffraction spectral separation imaging.RSC advances · 2022Article
- Simultaneous Multicolor DNA-PAINT without Sequential Fluid Exchange Using Spectral Demixing.Nano letters · 2022Article
- Live-cell fluorescence spectral imaging as a data science challenge.Biophysical reviews · 2022Review
- Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells.Communications biology · 2022Article
- Multiplexed structured illumination super-resolution imaging with lifetime-engineered upconversion nanoparticles.Nanoscale advances · 2021Article
- Article
- Completing the canvas: advances and challenges for DNA-PAINT super-resolution imaging.Trends in biochemical sciences · 2021Review
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recent advancements in single-molecule-based superresolution microscopy have made it possible to visualize biological structures with unprecedented spatial resolution. Determining the spatial coorganization of these structures within cells under physiological and pathological conditions is an important biological goal. This goal has been stymied by the current limitations of carrying out superresolution microscopy in multiple colors. Here, we develop an approach for simultaneous multicolor superresolution imaging which relies solely on fluorophore excitation, rather than fluorescence emission properties. By modulating the intensity of the excitation lasers at different frequencies, we show that the color channel can be determined based on the fluorophore's response to the modulated excitation. We use this frequency multiplexing to reduce the image acquisition time of multicolor superresolution DNA-PAINT while maintaining all its advantages: minimal color cross-talk, minimal photobleaching, maximal signal throughput, ability to maintain the fluorophore density per imaged color, and ability to use the full camera field of view. We refer to this imaging modality as "frequency multiplexed DNA-PAINT," or fm-DNA-PAINT for short. We also show that frequency multiplexing is fully compatible with STORM superresolution imaging, which we term fm-STORM. Unlike fm-DNA-PAINT, fm-STORM is prone to color cross-talk. To overcome this caveat, we further develop a machine-learning algorithm to correct for color cross-talk with more than 95% accuracy, without the need for prior information about the imaged structure.
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