Course Outline: The Immunohistochemistry and Microscopy (IHCM) course is four full days and evenings (11 hours daily) of lecture and laboratory sessions with experts in the field of immunohistochemistry (IHC) and microscopy. The IHCM course goal is to provide participants in-depth theory of and extensive hands-on experience with immunohistochemistry (IHC) techniques as well as theory and hands-on experience with a broad range of microscopic imaging techniques. The course emphasizes hands-on laboratory time and small breakout discussions with faculty and staff.
The laboratory demonstrations, exercises, discussions, and trouble-shooting sessions focus on: 1) Principles underlying the fixation of proteins in tissues. Because fixation and preparation of tissue prior to IHC constitutes one of the most important processes affecting the success of IHC studies, students will learn about and have multiple hands-on experiences with tissue preparation; they will also learn how to trouble shoot problems; 2) Antigen retrieval. Students will learn about and discuss how the detection of many antigens can be significantly improved by antigen retrieval. 3) Controls. Because IHC experiments must include positive and negative controls to support the validity of staining and identify experimental artifacts, students will learn about, discuss, and use controls that support the specificity of IHC results as well as the variances in antibody specificity and conditions that may generate inconsistent immune-staining and lead to inaccurate conclusions; 4) Strategies for detecting the presence of specific antigens in cells. Students will learn about and use antibodies labeled with different fluorescent probes and analyze the results with epi-fluorescence or confocal microscopies; they will also use antibody dilution studies; 5) Technologies that automate immunohistochemistry. To achieve a better understanding of when and how new technologies will benefit their research, students will be introduced to and use automated immunohistochemistry. 6) Basic elements of light and fluorescence microscopies. To gain insight into how to choose the correct imaging platform for their samples, students will learn to use various imaging platforms, including bright-field, epi-fluorescence, wide-field deconvolution, and confocal microscopies. Using images collected in the course as well as previously acquired images, students will also learn about and discuss acceptable practice for image capture and management and appropriate use of Photoshop to develop figures for publication; and 7) Troubleshooting. How to troubleshoot problems with immunohistochemistry and images will be an integral part of the course; troubleshooting sessions will be held daily.
This course is appropriate for undergraduate and graduate students, laboratory technicians, postdoctoral students, new and established faculty/clinicians seeking to expand their techniques and knowledge of IHC and microscopy. It is appropriate for beginning scientists and those with more advanced skills. Participants will be grouped appropriately. Registration is limited to thirty.
Support is provided by:
Bitplane/Andor TechnologyCarl Zeiss MicroImaging, LLC
Histochemical Society (course founder)
Journal of Histochemistry and Cytochemisty
SPOT Imaging Solutions, a division of Diagnostic Instruments, Inc.
2014 Academic and Industry Course Faculty:
Denis Baskin, University of Washington School of Medicine
Charles Frevert, University of Washington School of Medicine
Matthew Gastinger, Bitplane/Andor Technology
Paul Goodwin, Applied Precision
Stephen M. Hewitt, National Cancer Institute
Rebecca L. Hull, University of Washington
Jim McIlvain, Carl Zeiss MicroImaging, LLC
Philip Merlo, SPOT Imaging Solutions, a division of Diagnostic Instruments, Inc.
Sara Phinney, Leica Biosystems
Eduardo Rosa-Molinar, University of Puerto Rico-Rio Piedras
William L. Stahl, University of Washington School of Medicine
2014 Course Technical Faculty:
Brian Johnson, University of Washington School of Medicine
Rosedelma Diaz-De Leon, University of Puerto Rico-Rio Piedras
Noraida Martinez-Rivera, University of Puerto Rico-Rio Piedras
Irma I. Torres-Vazquez, University of Puerto Rico-Rio Piedras
Jose L. Serrano-Velez, University of Puerto Rico-Rio Piedras