 | Real-Time Interactive Lecture, Laboratory Protocol and Discussion |
 | Team taught by active researchers |
 | Lectures, Lab Protocols, Reference Materials included |
 | Space limited to 24 participants |
 | No Travel, Convenient and Cost Effective |
Course Director
Laboratory Demonstration, Protocol Review:
Preparation of Samples for Multicolor Analysis Flow Sorting; Multicolor Analysis, Compensation and Flow Sorting
Laboratory Demonstration:
a. BD Accuri C6:
Staining of Human Peripheral Blood Mononuclear Cells (PBMC) (hands on for each participant) with Basic Flow Panel (4 colors). Flow Cytometer Setup (BD Accuri; two lasers) with Special Emphasis on Compensation. Basics on Flow Cytometer Settings - Voltages, Gating, Dump Channels, Exclusion, and Compensation.
b. Miltenyi MACSQuant Analyzer 10
Advanced Flow Cytometry Laboratory
Analyzing human whole blood and human serum. This rotation includes: Automatic instrument calibration, automatic 8 color compensation, acquisition of whole blood with no RBC lysis, automatic gating/analysis of 8 color immunophenotyping panel, introduction to cytokine multiplex assay, acquisition of cytokine multiplex assay, automatic gating/analysis of cytokine multiplex assay, and automatic shutdown of instrument. In addition to immunophenotyping and multiplex assay, rare cell enrichment and cell counting will be discussed.
c. Bio-Rad S3e Flow Sorter
Emphasis on Sorting Conditions (parameters, factors, strategies) as well as the Specific Challenges Related to Cell Preparation, Efficiency, Recovery, and Alternate Techniques for Cell Population Identification.
d. Sony SH800 Flow Sorter
Single Cell Deposition
In Silico Software Training
a. Introduction to FlowJo Cytometry Analysis Software
b. Introduction to Cyflogic Cytometry Analysis Software; Flow Cytometry - Acquisition/Trouble Shooting/ Software
c. Introduction to FlowLogic Cytometry Analysis Software
d. Introduction to FCS Express Cytometry Analysis Software
Elke Bergmann-Leitner, Ph.D.
Dr. Elke Bergmann-Leitner is the Chief of the Flow Cytometry Center (Malaria Vaccine Branch) at the Walter Reed Army Institute of Research in Silver Spring, MD. She is an adjunct Research Associate Professor of Medicine at the Uniformed Services University of Health Sciences and has been teaching flow cytometry for 25 years. Currently, her main research focus is the development of readout methods capable of identifying immune correlates of protection against infectious diseases.
Wolfgang Leitner, Ph.D.
Dr. Leitner is the Chief of the Innate Immunity Section at the National Institute of Allergy and Infectious Diseases (NIH/NIAID). He received his flow cytometry training at the University of Salzburg where he obtained a MSc in biochemistry and a PhD in Immunology. He has been teaching flow cytometry for 25 years. Before joining NIAID, he worked on melanoma and lymphoma vaccines at NCI/NIH, and at WRAIR as a vaccine consultant.
