Course Outline

Topics Covered in Hands On Immunohistochemistry Forum
9:00am - 4:30pm

Pretreatment 
•Describe what pretreatment is and why it is performed 
•Explain the different pretreatment methodologies, their uses and limitations
   o Enzyme unmasking
   o Heat unmasking
   o Chemical unmasking

Antibodies
•Summarize how immune reactions are used to produce antibodies (anti-sera) used in Immunohistochemistry (IHC) techniques
•Give a basic explanation of the various production methods for antibodies
   o Monoclonal
   o Polyclonal
   o Rabbit Monoclonal
•Classify the various antibody formats and make use of a specification sheet
•Explain how titrations are used in immunohistochemistry

Secondary Antibodies
•Explain the difference in a secondary antibody and a primary antibody
•Discuss how secondary antibodies are created for immunohistochemistry assays
•Identify the approach you would use to select the best secondary antibody for an immunohistochemistry assay/stain

Detection
•Discuss the concept of detection chemistry
•Identify the types of "labels" used for detection:
   o Fluorescent & Chromogenic labels
•List the differences between the types of detection technologies, their uses, and limitations

Counter Staining
•Explain who counterstaining is important to an immunohistochemistry stain
•Summarize in your own words some of the ways mounting media can impact immunohistochemistry staining

Double Staining
•Differences between Single/Double Staining

Multiplexing
•Understand why multiplexing is important
•Explain key components of running a multiplex immunohistochemistry stain
•Summarize the importance of running proper drop-out controls during denaturation

Background Elimination-  Blockers
•Articulate what blockers are and what they are used for
•Discuss potential sources of non-specific staining
•Differentiate between the types of blockers, their uses, and limitations to include:
   o Protein blockers
   o Enzyme blockers
   o Biotin blockers
   o Fc blocker

Troubleshooting
•Artifacts
•Automation  

Meet the Faculty

Theresa Burchette, MTL, HT(ASCP), Technical Business Manager, Dako, an Agilent Technologies Company

Theresa received her Bachelor of Science degree in Medical Technology from Upstate Medical Center in Syracuse, New York. She is an ASCP registered Medical Technologist & Histologist. She has worked for Dako, an Agilent Technologies Company for over 9 years currently serving as a Technical Business Manager, covering the East Region of North America. She has covered Northern Florida as an account manager and has worked with other sales managers within Dako as a business manager as well as a global trainer. Prior to Dako, Theresa developed the RNA extraction and amplification reagent product line for the Laser Capture Microdissection company, Arcturus and assisted customers with their specific applications. She also worked for Glaxo SmithKline in Genomic Histology, where she provided histology support for the entire company with a focus on gene target localization and expression profiling through IHC and ISH. She worked at Applied Biosystems in the Science and Technology group, developing future technologies for the company. She worked as a Histologist as well as started a molecular laboratory in a reference hospital, setting up over 30 molecular assays. Theresa has more than 25 years of technical expertise and is the author of several publications and patents.

Richard Cartun, MS, PhD, Director, Histology/Morphologic Proteomics, Hartford Hospital

Dr. Cartun currently serve as the Director of Histology, Immunopathology, and the Martin M. Berman, MD Morphologic Proteomics Laboratory at Hartford Hospital, a large tertiary care hospital in Hartford, CT. He has been doing immunohistochemical testing for more than 35 years now and, as a result, has extensive knowledge in the technical aspects of this critically important laboratory test.

Traci DeGeer, HT/HTL(ASCP)QIHC , Discovery Training Manager, Roche

Traci DeGeer is the Discovery Training Manager for Roche Molecular Systems where she develops training programs researchers and Roche training sites world wide.. received her bachelors of science in Biology from Radford University in Virginia. She is certified both as a HT and a HTL and has her QIHC. At the start of her career she worked in the clinical setting as a Histotechnologist, Lead Histotechnologist. She then moved to Ventana Medical Systems in 2004 as a technical customer care executive and 8 months later into training. has performed numerous workshops at both the state and national level on a variety of different topics. is an active member of the NSH where she serves as chair of the Education and CEU committees.

Brent Heller, Technical Consultant, Millipore Sigma

Brent received his bachelors degree in Biology from California State University -Long Beach. He has 6 years working in IHC and histology and through his work as a Technical Consultant with Millipore Sigma he has been fortunate enough to work with and visit several labs around the United States. Brent is a frequent and highly rated presenter at state and national conventions as well as webinars. He is currently involved with IHC in the southern part of the United States and believes strongly in always continuing his own education in histology.

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