Super Resolution Microscopy represents a group of recently developed light microscopic techniques that are able to exceed diffraction-limited resolution (less than 200nm). This course will focus on three types of Super Resolution Microscopy- Structured Illumination Microscopy (SIM), Stochastic Optical Reconstruction Microscopy (STORM), and Stimulated Emission Depletion (STED). In addition, students will be exposed to cutting-edge super resolution microscopes developed at HHMI Janelia Research Center through the AIC (Advanced Imaging Center). The AIC will showcase several instruments, including iPALM, lattice light sheet and live-cell TIRF-SIM.
The course is designed for cell biologists with prior experience in light microscopy who wish to add super resolution microscopy to their research program. Participants will acquire both theoretical understanding of super resolution microscopy and practical experience using state-of-the-art super resolution microscopes.
Course/Workshop Location
March 15-17, 2016: NIH Main Campus (Bethesda)
March 18, 2016: HHMI Janelia campus (Complimentary transportation will be provided to and from the HHMI Janelia Farm campus.)
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Lecture Topics
Introduction to Super Resolution Microscopy; Advances in Super Resolution Microscopy; Theoretical Background of SIM, PALM/dSTORM, and STED Imaging; Advances in Fluorescent Protein and Organic Dye Technology; Applications of Super Resolution Microscopy using SIM and dSTORM; Challenges Associated with Obtaining Good SIM, STORM and STED Images; Potential Artifacts Common to Super Resolution Imaging.
Laboratory Topics
Working in groups of five, participants will rotate through multiple work stations.
The following work stations are located on NIH campus:
- Leica gSTED, GE- OMX SIM, Nikon N-STORM, Zeiss Elyra PALM/SIM, Zeiss LSM880-AiryScan, Shroff Lab - Instant SIM, Shroff Lab - Dual View Plane Illumination Microscope.
Confirmed Speakers/Instructors
Drs. Hari Shroff, Jennifer Lippincott-Schwartz, Clare Waterman, Jordan Beach, Srich Murugesan, Wesley Legant, George Patterson, Xufeng Wu, Jason Yi, Liang Gao, Kem Sochacki, Chris Combs, Lin Shao, Leong Chew
Another three stations are located on Janelia Farm campus including iPALM, Lattice light sheet microscope, TIRF-SIM for live cell imaging. Complimentary transportation will be provided to and from the HHMI Janelia Farm campus.
During the laboratory sessions, the following topics will be covered:
- Introduction and Feature Highlights of the Instrument
- Imaging Acquisition Procedures
- Sample Preparation Requirements and Recommendations (Dye Choices and Imaging Solutions)
- Imaging Data Analysis and Presentation
- Trouble-shoot Common Problems in SIM Imaging
- Acquisition of 2D and 3D STORM Images