Rhodamine spiroamides for multicolor single-molecule switching fluorescent nanoscopy

Area of Science:

• * Chemistry
• * Biophysics
• * Nanotechnology

Background:

• * Rhodamine spiroamides are fluorescent molecules with potential applications in advanced microscopy.
• * Single-molecule localization microscopy (SMLM) requires probes with specific photoactivation and switching properties.
• * Precise determination of molecular positions and shapes is crucial in biological studies.

Purpose of the Study:

• * To design, synthesize, and evaluate novel rhodamine spiroamides for SMLM.
• * To develop fluorescent probes with enhanced photoactivation, multicolor capabilities, and selective binding.
• * To demonstrate the application of these probes in nanoscale imaging and colocalization studies.

Main Methods:

• * Chemical synthesis of novel rhodamine spiroamide derivatives.
• * Characterization of photophysical properties, including photoactivation and fluorescence.
• * Evaluation of probe performance in single-molecule switching and localization microscopy experiments.
• * Application in colocalization studies of biological objects using two-photon activation.

Main Results:

• * Successful design and synthesis of new rhodamine spiroamides with tunable properties.
• * Achieved multicolor staining, good photoactivation, and high photon emission.
• * Demonstrated selective chemical binding to amino or thiol groups.
• * Achieved nanoscale resolution (tens of nanometers) in optical imaging and colocalization studies.

Conclusions:

• * The developed rhodamine spiroamides are effective fluorescent probes for SMLM.
• * These probes facilitate precise nanoscale imaging and colocalization of biological structures.
• * The combined switching and fluorescent groups in a single molecule offer a promising platform for future SMLM development.