Single Molecules Are Your Quanta: A Bottom-Up Approach toward Multidimensional Super-resolution Microscopy

Area of Science:

• Biophysics
• Nanotechnology
• Microscopy

Background:

• Single-molecule localization microscopy (SMLM) and super-resolution techniques have advanced biological and material science imaging.
• The accumulation of single-molecule measurements allows for the creation of diffraction-unlimited images.

Purpose of the Study:

• To reflect on the philosophy and opportunities of building nanoscale images molecule by molecule.
• To highlight emerging extensions of SMLM into new dimensions and functionalities.

Main Methods:

• Analysis of the bottom-up accumulation approach in SMLM.
• Introduction to established multicolor and 3D SMLM.
• Highlighting emerging SMLM functionalities like fluorescence polarization, emission spectra, and molecular motion tracking.

Main Results:

• SMLM allows for the gradual emergence of detailed spatial and functional information.
• Established multicolor and 3D SMLM provide enhanced imaging capabilities.
• Emerging functionalities promise further expansion of SMLM's potential.

Conclusions:

• The bottom-up accumulation of single molecules as quanta is a powerful method for multidimensional nanoscale microscopy.
• SMLM offers unique opportunities for detailed investigation of complex systems.
• Future directions point towards integrating more dimensions and functionalities into SMLM.