IMMUNOCYTOCHEMISTRY, IN SITU HYBRIDIZATION, SUPER-RESOLUTION & LIVE CELL IMAGING
October 13 - 26, 2010
Application Deadline: July 15, 2010

Instructors:
Viki Allan, University of Manchester, UK
Ke Hu, Indiana University
John Murray, University of Pennsylvania

a nice collage from a recent course

This course focused on specialized techniques in microscopy, in-situ hybridization, immunocytochemistry, and live cell imaging related to localizing DNA, RNA, and proteins in fixed cells as well as protein and RNA dynamics in living cells. The course emphasized the use of the latest equipment and techniques in fluorescence microscopy, including: confocal laser scanning microscopy; deconvolution methods; several super-resolution methods including structured illumination, STORM, and PALM; digital image processing, and timelapse imaging of living specimens. The course was designed to present students with stateof-the-art technology and scientific expertise in the use of light microscopy to address basic questions in cellular and molecular biology. The course was designed for the molecular biologist who is in need of microscopic approaches and for the cell biologist who is not familiar with the practical application of the advanced techniques presented in the course. Among the methods presented were: the preparation of tagged nucleic acid probes; fixation methods; detection of multiple DNA sequences in single nuclei or chromosome spreads; cellular localization of RNA; localization of nucleic acids and proteins in the same cells; use of a variety of reporter molecules and non-antibody fluorescent tags; indirect antibody labeling; detection of multiple proteins in a single cell; labeling antibodies with two fluorophores for STORM; the use of GFP variants to study protein expression in living cells by conventional microscopy and PALM; use of photo-activatable and photo-switchable fluorescent proteins for studying localization and dynamics. In each method, several experimental protocols were presented allowing the students to assess the relative merits of each and to relate them to their own research. Students were encouraged to bring their own nucleic acid, protein, or antibody probes to the course, which were used in addition to those provided by the instructors. The laboratory exercises were supplemented with lectures given by invited distinguished scientists, who presented up-to-the-minute reports on current methods and research using the techniques being presented.

Speakers in the 2009 course included:
Mark Bates, Harvard University
Richard Day, Indiana University School of Medicine
Alison North, Rockefeller University
Thomas Ried, National Cancer Institute/NIH
Hari Shroff, National Institutes of Health
David Spector, Cold Spring Harbor Laboratory
Xiaowei Zhuang, Harvard University


This course is supported with funds provided by the National Cancer Institute