Simultaneous co-localized super-resolution fluorescence microscopy and atomic force microscopy: combined SIM and AFM...

Area of Science:

• Cellular Biology
• Microscopy
• Biophysics

Background:

• Correlating data from diverse microscopy techniques can reveal novel cellular signaling events.
• Previous simultaneous imaging approaches were limited by the Abbe diffraction limit in fluorescence microscopy.

Purpose of the Study:

• To develop and demonstrate a hardware setup for simultaneous, co-localized imaging using super-resolution fluorescence microscopy and atomic force microscopy.
• To overcome the spatial resolution limitations of traditional fluorescence microscopy.

Main Methods:

• Integration of a hardware setup for simultaneous far-field super-resolution fluorescence microscopy and atomic force microscopy.
• System performance validation using sub-resolution fluorescent beads.
• Application to human bone osteosarcoma epithelial cells expressing N-terminally EGFP-tagged MCT1.

Main Results:

• Achieved simultaneous co-localized imaging with spatial correlation between super-resolution fluorescence and atomic force microscopy.
• Demonstrated system capability beyond the Abbe diffraction limit.
• Successfully imaged plasma membrane transporter 1 (MCT1) in human cells.

Conclusions:

• The developed hardware enables unprecedented correlated imaging, advancing cellular biology research.
• This technique overcomes previous resolution limitations, opening new avenues for studying cellular signaling.
• The system provides a powerful tool for high-resolution, multi-modal cellular imaging.