Saturday, March 17: Arrival and registration.
Sunday, March 18: Course begins and continues through Tuesday, March 20th;
The following topics will be covered in course. Topics with a lab component are noted.
AMPLIFICATION METHODS – tyramide signal amplification is a powerful enzymatic technique for enhanced IHC detection. It provides increased sensitivity and requires very low concentrations of antibodies. Lab module included
ANTIGEN RETRIEVAL – method that changed the practice of IHC. Use of this method on deparaffinized, formalin-fixed tissues gives improved signal, reduced noise, use of higher antibody titers and use of antibodies not otherwise applicable on formalin-fixed tissue. The method is especially useful for basic research and for diagnostic pathology. Lab module included.
CONTROLS – several basis controls are essential to help support interpretation of results in IHC. Discussion of what you need to do to help validate your work.
DOUBLE LABELING AND CO-LOCALIZATION – strategies for detecting the presence of specific antigens in cells. This is typically done using antibodies labelled with different fluorescent probes and analyzed either by epifluorescence or confocal microscopy. How do you determine that two different proteins are present in the same compartment or cell? Antibody dilution studies are essential for successful outcomes.
FIXATION – principles underlying the fixation of proteins in tissues. Fixation and preparation of tissue prior to IHC are one of the most important processes that will affect success for IHC studies.
LIGHT AND FLUORESCENT MICROSCOPY – basic concepts essential for carrying out light and fluorescent laboratory work in the course. Tutorial on use of light and fluorescent microscopes.
IMAGE AUTHENTICITY – What is acceptable practice for image capture and management? This includes archiving, use of Photoshop techniques and related topics. Tutorial on how to use Photoshop to analyze image stacks from confocal microscopy images for co-localization of proteins. Images collected in the course as well as previously acquired images will be used.
TROUBLESHOOTING – This is integrated with laboratory sessions which are part of the various modules within the course. What might be wrong when you don’t get an expected result. Additionally, the course will end with a session in which a panel of faculty will discuss troubleshooting.
MICROWAVE PROCESSING – The use of modern microwave systems speeds IHC procedures significantly. For example, a two-day procedure may be reduced to several hours with superior results at significant savings of time and effort. Lab module included.
