Correlative Super-resolution Optical and Electron Microscopic Imaging of Intracellular Ribosomal RNA by a Terpyridine...

Area of Science:

• Molecular Biology
• Cell Biology
• Bioimaging

Background:

• Ribosomal RNA (rRNA) is crucial for protein synthesis, but visualizing its live-cell dynamics at the nanoscale remains challenging.
• Existing tools lack the capability for multimodal, high-resolution imaging of intracellular rRNA distribution.
• Understanding rRNA's role requires advanced techniques for observing its behavior during active biological processes.

Purpose of the Study:

• To design and synthesize novel iridium(III) complexes for selective live-cell labeling of rRNA.
• To develop a nanoscale imaging tool for visualizing rRNA distribution and dynamics.
• To investigate the correlation between rRNA visualization, protein synthesis, and cellular structures.

Main Methods:

• Rational design and synthesis of terpyridine ammonium iridium(III) complexes.
• Selective labeling of rRNA in living cells using a developed iridium(III) complex.
• Super-resolution STED imaging for high-resolution rRNA visualization.
• Correlative light and electron microscopy (CLEM) for multimodal imaging.
• Assessment of rRNA dynamics and impact on protein synthesis.

Main Results:

• A specific iridium(III) complex selectively labeled rRNA in living cells.
• Achieved nanoscale imaging of rRNA on the rough endoplasmic reticulum with ~40 nm resolution via STED microscopy.
• Demonstrated correlation between optical and electron microscopy findings using CLEM.
• Observed rRNA dynamics in living cells and noted a boost in nanoscale protein synthesis.

Conclusions:

• The developed iridium(III) complex is a versatile tool for multimodal, nanoscale visualization of rRNA in live cells.
• This technique provides new insights into rRNA dynamics and its role in protein synthesis.
• The study advances the understanding of rRNA's function and behavior within living systems.