I. Intracellular Fluorescent Imaging: An Introduction
II. Using Photobleaching and Photoactivation to Study the Endomembrane System
III. Super Resolution Imaging
Part II: Using Photobleaching and Photoactivation to Study the Endomembrane System
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Recent breakthroughs in intracellular fluorescent imaging allow the visualization, tracking, and quantification of molecular interactions within living cells and whole organisms. In part 1 of this talk, Lippincott-Schwartz gives an overview of the development of fluorescent protein markers, explains how these proteins are used to label intracellular compartments, and how fluorescent microscopy is used to follow events within cells. In the second lecture, Lippincott-Schwartz describes how two imaging techniques, photoactivation and photobleaching (FRAP), can be used to switch on or off specific subsets of fluorescent molecules. She and others have used these techniques to ask questions about the kinetics of the movement of specific molecules through the secretory pathway. The third lecture focuses on super-resolution imaging, or Photo Activated Localization Microscopy (PALM), a process that allows the behavior of individual fluorescent molecules to be followed.
Dr. Lippincott-Schwartz is Chief of the Section on Organelle Biology in the Cell Biology and Metabolism branch of the National Institutes of Health. Using a variety of fluorescent imaging techniques in live cells, Dr. Lippincott-Schwartz and her lab study dynamic protein interactions within cells, in real time and space. Her studies span a range of topics including organelle assembly and disassembly, the cytoskeleton and protein transport and the establishment of cell polarity.
After receiving her BA from Swarthmore College, Dr. Lippincott-Schwartz taught for several years (including a few years in Kenya) before returning to the lab to receive a Master’s degree from Stanford and her PhD from Johns Hopkins University. She did her post-doctoral work at the NIH with Richard Klausner. Dr. Lippincott-Schwartz is a member of the National Academy of Sciences and is active on numerous editorial boards.
- Jennifer Lippincott-Schwartz iBioMagazine: How Do Lipids and Cholesterol Regulate Trafficking Across the Secretory Pathway
- Martin Chalfie iBioMagazine: Developing GFP as a biological marker
- Roger Tsien iBioEducation Lecture: Fluorescent Proteins
- Xiaowei Zhuang iBioSeminar: Fluorescent imaging at nanoscale
- Eric Betzig and Harald Hess iBioMagazine: Developing PALM microscopy
Patterson, G., Hirschberg, K., Polishchuk, R.S., Gerlich, D., Phair, R.D. and Lippincott-Schwartz, J. (2008) Transport through the Golgi Apparatus by Rapid Partitioning within a Two-Phase Membrane System. Cell 133:1055-1067.
Kim, P.K., Hailey, D.W., Mullen, R.T. and Lippincott-Schwartz, J. (2008) Ubiquitin signals autophagic degradation of cytosolic proteins and peroxisomes. PNAS 105:20567-20574.
Lippincott-Schwartz J. and Manley, S. (2009) Putting super-resolution fluorescence microscopy to work. Nature Methods 6:21-23.
Lippincott-Schwartz, J. and Patterson, G. (2009) Photoactivatable fluorescent proteins for diffraction-limited and super-resolution imaging. Trends Cell Bio. 19:555-564.
Patterson, G., Davidson, M., Manley, S. and Lippincott-Schwartz, J. (2010) Superresolution Imaging using Single-Molecule Localization. Annu. Rev. Phys. Chem. 61:345-67.