Three-dimensional super-resolution imaging of live whole cells using galvanometer-based structured illumination...

Area of Science:

• Cellular and Molecular Imaging
• Biophysics
• Microscopy Techniques

Background:

• Live-cell imaging of 3D nanoscale organization is crucial but challenging.
• 3D structured illumination microscopy (SIM) offers advantages for live cells due to wide-field imaging and no need for high light intensity or special dyes.
• Development of 3D SIM systems, particularly for live imaging, has been slow.

Purpose of the Study:

• To report a flexible and high-speed 3D SIM system for live-cell dynamics.
• To overcome limitations in current 3D SIM systems for live imaging.
• To enable broader applications of SIM in life sciences.

Main Methods:

• Development of a 3D SIM system utilizing two galvanometer sets.
• Control of structured illumination pattern's period and orientation using galvanometers.
• Demonstration of capabilities through imaging nanoparticles, microtubule organizations, and mitochondria evolution.

Main Results:

• Achieved strong optical sectioning.
• Obtained lateral resolution of 104 nm, axial resolution of 320 nm.
• Reached volume temporal resolution of 4 seconds for live-cell dynamics.
• Demonstrated imaging of nanoparticle and microtubule organizations and mitochondria evolution.

Conclusions:

• The galvanometer-based 3D SIM system offers enhanced flexibility and speed for live-cell imaging.
• This system broadens the imaging capabilities of SIM microscopes.
• Facilitates advanced applications in life sciences research by enabling real-time study of cellular dynamics.