Multiscale spectral nanoscopy

A method for non-invasively tracking a particle in a sample, comprising:

• providing a 2-photon or confocal laser scanning microscope (LSM) with a plurality of scanning mirrors and a plurality of detectors configured to scan the sample;
• providing a particle-holding device with the particle coupled to the particle-holding device, the particle-holding device being coupled to a stage with X-Y and Z position control;
• providing a tracking module configured to:
• generate a tracking excitation laser beam;
• detect deviations of the particle in an X-Y and Z direction;
• and
• generate control signals to drive the stage X-Y and Z position controls to track movement of the particle;
• and synchronizing the LSM and tracking module by:
• receiving current scanning mirror positions and detector readouts from the LSM;
• receiving a current stage position from the tracking module;
• and
• generating an LSM pixel stamped with a stage position.

A system for non-invasively tracking a particle in a sample, comprising:

• an imaging module configured to scan the sample comprising:
• a laser scanning microscope (LSM);
• a plurality of scanning mirrors;
• and
• a plurality of single point detectors;
• a microscope stand comprising:
• a sample holder containing the sample;
• and
• a stage configured to provide X-Y and Z position control;
• a tracking module comprising:
• a tracking excitation laser;
• X-Y radiation-gathering components configured to detect deviations of the particle in an X-Y direction;
• Z radiation-gathering components configured to detect deviations of the particle in a Z direction;
• and
• a processor configured to generate control signals to drive the stage X-Y and Z position controls to track movement of the particle;
• and
• a synchronization module configured to synchronize the imaging module and tracking module by:
• receiving current scanning mirror positions and detector readouts from the imaging module;
• receiving a current stage position from the tracking module;
• and
• generating an LSM pixel stamped with a stage position.