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Unraveling cellular complexity with transient adapters in highly multiplexed super-resolution imaging
Florian Schueder1, Felix Rivera-Molina2, Maohan Su2
1Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA; Department of Microbial Pathogenesis, Yale School of Medicine, New Haven, CT, USA.
Cell
|March 29, 2024
Summary
Researchers developed FLASH-PAINT, a rapid multiplexed super-resolution microscopy technique. This method enables imaging of numerous molecular targets, advancing cellular and molecular biology research.
Area of Science:
- Cellular Biology
- Microscopy
- Molecular Biology
Background:
- Understanding cellular functions requires mapping molecular spatial relationships.
- Super-resolution microscopy provides high resolution but is limited to few simultaneous targets.
- Multiplexed imaging with sequential labeling is low-throughput.
Purpose of the Study:
- To develop a rapid, high-throughput multiplexed super-resolution microscopy method.
- To enable imaging of a nearly unlimited number of molecular targets.
- To overcome limitations of current super-resolution techniques for complex cellular mapping.
Main Methods:
- Developed FLASH-PAINT (Fluorogenic Labeling and Transient Adapter-Mediated Switching for High-throughput DNA-PAINT).
- Utilized fluorogenic labeling and transient adapter-mediated switching.
- Applied DNA-PAINT for super-resolution imaging.
Main Results:
- Demonstrated FLASH-PAINT versatility across four applications.
- Mapped nine proteins in a single mammalian cell.
- Elucidated primary cilia organization, Golgi stack dynamics (13 targets), and inter-organelle contacts in 3D super-resolution.
Conclusions:
- FLASH-PAINT significantly enhances throughput for multiplexed super-resolution microscopy.
- The method allows for detailed investigation of complex molecular interactions and cellular structures.
- FLASH-PAINT offers a powerful tool for advancing cellular and molecular biology research.
