Revolutionizing cellular imaging with FLASH-PAINT

12-target FLASH-PAINT imaging of Golgi complexes in untreated and nocodazole-treated HeLa cells. Images generated (Schueder F., et al., 2024).

From DNA-PAINT to FLASH-PAINT

FLASH-PAINT (Fluorogenic Anchored Probes with DNA Hybridization-PAINT) builds on the DNA-PAINT technique (Jungmann, R., et al., 2010; Schoen, I., et al., 2012). DNA-PAINT utilizes transient binding of fluorescent DNA probes to complementary DNA strands anchored to target molecules, achieving high-resolution imaging. However, its throughput is limited due to the need for separate probes for each target. FLASH-PAINT, developed at Yale by the laboratory of Joerg Bewersdorf, PhD., (Schueder F., et al., 2024) overcomes this by using flexible adapters that transiently bind to targets, allowing for the rapid and multiplexed imaging of numerous molecular features simultaneously.

Key Advantages

• Unlimited multiplexing: FLASH-PAINT can image countless proteins and other molecular features simultaneously, providing a comprehensive view of cellular processes.

• Speed and efficiency: The technique is 100 times faster than current super-resolution methods, reducing the time and cost involved in cellular imaging.

• High resolution: It achieves nanometer-scale resolution, essential for detailed mapping of cellular structures and interactions.

Applications in Spatial Biology

Spatial biology seeks to understand the organization and interactions of molecules within their spatial context in cells and tissues. FLASH-PAINT’s ability to visualize multiple targets makes it a powerful tool for:

• Mapping protein localization: Detailed spatial maps of protein distribution can elucidate cellular architecture and function.

• Studying dynamic processes: Real-time tracking of molecular interactions provides insights into cellular mechanisms.

• Investigating diseasemechanisms: Visualizing the spatial arrangement of biomolecules can reveal the underlying causes of diseases, aiding in the development of targeted therapies.

Real-World Impact

In their study, Yale researchers used FLASH-PAINT to map nine proteins in a single mammalian cell, analyze the organization of primary cilia, and investigate inter-organelle contacts in 3D. This level of detail was previously unattainable, opening new avenues for understanding cellular functions and disease processes.

As Joerg Bewersdorf, PhD, one of the lead researchers, explains, “Instead of doing a hundred experiments looking at individual interactions of one or two proteins, we can now do a single experiment where we can see all possible interactions” (Yale Medicine article).

The Future of Spatial Biology

The potential applications of FLASH-PAINT in spatial biology are vast. Future research may extend its use to tissue imaging and diagnostics, providing clinicians with new tools to visualize complex cellular environments and improve disease treatments. With its high-throughput, cost-effective, and detailed imaging capabilities, FLASH-PAINT stands to revolutionize our approach to studying the intricate world within cells.

For further details, check out the original Yale Medicine article and the comprehensive study published in Cell (Schueder F., et al., 2024).

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#Microscopy #SuperResolution #SpatialBiology