Advances in Optical Microscopy: From Single Molecules to Four-Dimensional Imaging
BCH 2127H
Contemporary optical microscopy plays a central role in modern cell biology and biophysics. A variety of techniques have been developed to visualize and track organelles and even single molecules in live cells and organisms, and dedicated algorithms enable sophisticated analysis of the localization, shape, motion and other properties of defined structures. Recent developments have improved the resolution of optical microscopy below the diffraction limit that hampered fluorescence microscopy for decades. Moreover, allied techniques now enable the assessment of molecular diffusion, proximity, rotational motion, force etc., as well as cellular and organellar properties such as viscosity, tension and surface charge. The course will describe and discuss topics including single-molecule tracking, several modes of super-resolution microscopy, photobleaching and photoactivation, fluorescence lifetime imaging, energy transfer, optical tweezers, multiphoton intravital microscopy, etc.
Topics BCH2127H (2023)
January 13
-General principles microscopy (SG) plus topic selection
January 20
-Quantum dots and SPT
-Use of fluorescent proteins to measure pH and calcium activity.
-TIRF (total internal reflection microscopy)
February 10
-Hyvolution/Lightning (Leica) and Airyscan (Zeiss) microscopy
-LLSFM (lattice light-sheet fluorescence microscopy)
-Multiphoton intravital microscopy/ the Femtonics system
February 17
-FLIM (fluorescence lifetime imaging: time and frequency domains); Polarization microscopy
-FRAP and FLIP (fluorescence recovery after photobleaching and fluorescence loss in photobleaching)
-FRET and BRET (fluorescence and bioluminescence energy transfer)
February 24
-PALM/STORM (photoactivated localization microscopy /stochastic optical reconstruction microscopy; photoconvertible fluorescent proteins)
-SIM (structured illumination microscopy)
-STED (stimulated emission depletion microscopy)/ -Minflux
March 3
-AFM (atomic force microscopy)
-CLEM (correlative light and electron microscopy)
-FCS (fluorescence correlation spectroscopy)
Non-Biochemistry Students: This courses will be available to all non-Biochemsitry students to request through ACORN as follows:
Winter 2023 modules: December 1, 2022 – January 23, 2023 (last day to add Winter courses)
**Important note for non-Biochemistry students – if you have one unpaired BCH2024 course, then you must contact carrie.harber@utoronto.ca to sign up for the second 1/4 module as per the deadlines listed above. Do not enroll through ACORN as your grades must be recorded under BCH 2024H Y – Focused Topic in Biochemistry.
Course Next Offered
Winter 2023
Course Time and Location
Start Date: January 13, 2023
End Date: March 3, 2023
Time: 3-5 pm
Day(s) of the Week: Friday
Location: Medical Sciences Building, Room 5231
Enrollment Limit
Yes — 15
Method of Student Evaluation
Students will be evaluated based on their assigned presentation and general participation in discussions.
Coordinator
Spencer A. Freeman
SickKids Research Institute
Peter Gilgan Centre for Research and Learning
686 Bay Street
Room 18.9709
416-813-7654, ext. 309267
spencer.freeman@sickkids.ca