Course at a Glance
Far field fluorescence super-resolution microscopy, from theory to applications. The course will be focused on the concept behind fluorescence super-resolution microscopy with particular attention to single molecule localization techniques (SML). Furthermore basics principles of structured illumination SIM, RESOLFT, STED and Expansion microscopy will be also explained.
Instructor
Francesca Cella Zanacchi francesca.cella@iit.it
Tel: +39 010 71 781 320
Nanophysics, Istituto Italiano di Tecnologia
Credits: 3
Synopsis
Breakthrough in optical microscopy and labeling techniques opened new possibilities in the investigation of protein organization and dynamics in biological systems. The recent development of super-resolution microscopy played a relevant role in the groundbreaking progress in life science and neuroscience. In particular, state-of-the-art far field super-resolution microscopy techniques based on single molecule localization not only provide
imaging capabilities with unprecedented resolution, but they also represent a powerful tool to quantify protein distribution and “count” molecules in biological system. This course will explain the basics of super-resolution techniques. Particular attention will be addressed to Single Molecule Localization Microscopy Techniques (SML) and Light sheet fluorescence microscopy. Basics principles of super-resolution techniques based (such as Structured illumination SIM, RESOLFT, STED and Expansion microscopy) will be also
explained.
Syllabus
The course develops in about 7 hours in the classroom and 2 hours of laboratory activity.
- Individual Molecule localization techniques: F-PALM, PALM, STORM.
- Single molecule Tracking
- Light sheet microscopy and super-resolution: SPIM, DSLM, Inclined Illumination and IML
-SPIM
- Structured Illumination microscopy (SIM) and saturated SIM (SSIM)
- Reversible Saturable Optical Fluorescence Transitions (RESOLFT) and stimulated emission microscopy STED
The examination consists in a journal club or a brief research project proposal.
Reading list
- Hell S.W., “Microscopy and its focal switch”, Nat. Methods, 6, 24-32 (2009)
- E. Betzig et al. (2006). "Imaging Intracellular Fluorescent Proteins at Nanometer Resolution". Science 313 (5793)
- Deschout, H., et al., Precisely and accurately localizing single emitters in fluorescence microscopy. Nat Methods. 11(3): p. 253-66 (2014).
- Francesca Cella Zanacchi et al. “Live-cell 3D super-resolution imaging in thick biological samples” Nature Methods 12/2011
- Vicidomini G. et al., “Sharper low-power STED nanoscopy by time gating”, Nat. Methods, 8, 571-573 (2011)
Venue
IIT - Via Morego, 30 16163 Genova
Course date
April 2017
Course date
June 2016
- Docente: FRANCESCA CELLA ZANACCHI