Thermal-plex: fluidic-free, rapid sequential multiplexed imaging with DNA-encoded thermal channels

Area of Science:

• Biotechnology
• Molecular Biology
• Microscopy

Background:

• Multiplexed fluorescence imaging is crucial for biological research but often limited by the number of detectable targets.
• Existing sequential imaging methods require complex fluidics and lengthy buffer exchange steps, hindering speed and accessibility.

Purpose of the Study:

• To introduce Thermal-plex, a novel fluidic-free method for rapid, high-plex sequential fluorescence imaging.
• To overcome the limitations of conventional sequential imaging techniques.

Main Methods:

• Developed simple DNA probes activated by specific temperature spikes (thermal channels) for sequential fluorescence activation.
• Utilized a commercially available heating device for rapid channel switching (<30 seconds).
• Performed imaging without buffer exchange or complex fluidic systems.

Main Results:

• Demonstrated 15-plex RNA imaging (5 thermal channels × 3 fluorescence channels) in fixed cells and retina tissues.
• Achieved complete 15-plex imaging in under 4 minutes.
• Showcased the speed and efficiency of the fluidic-free Thermal-plex approach.

Conclusions:

• Thermal-plex offers a significantly faster and simpler alternative for high-multiplexed sequential imaging.
• This method expands the capabilities of fluorescence microscopy for biological research.
• Introduced a new labeling strategy for efficient sequential multiplexed imaging.