Methods: Fundamentals of fluorescence microscopy and benefits of probing the sub-ensemble; Requirements and evidences for single-molecule/nanocrystal fluorescence detection; Advantages/drawbacks of various single-molecule (SM) imaging techniques; SM image processing, data analyses procedures and interpretation. Dynamics and Spectroscopy: Single-emitter fluorescence intermittency (blinking); SM diffusion (translational and rotational) dynamics; Spectroscopy of single emitters; SM fluorescence lifetime; single-emitter energy and electron transfer. Applications: Select applications of in chemistry, soft matter systems, biology/biophysics and nanomaterials science through discussion of seminal publications.

1. J. R. Lakowicz, Principles of Fluorescence Spectroscopy, 3rd Ed, Springer, 2007.

2. D. B. Murphy, Fundamental of Light Microscopy and Electronic Imaging, Wiley-LISS, 2001.

3. C. Gell, D. Brockwell and A. Smith, Eds. Handbook of Single Molecule Fluorescence Spectroscopy, Oxford, 2013.

4. M. Sauer, J. Hofkens and J. Enderlein, Eds. Handbook of Fluorescence Spectroscopy and Imaging: From Ensemble to Single Molecules, Wiley-VCH, 2010.

5. C. Brauchle, D.C. Lamb and J. Michaelis Eds. Single Particle Tracking and Single Molecule Energy Transfer, Wiley-VCH, 2010.