August 22-25, 2017

Tuesday - Friday

Germantown, MD

The Bioscience Education Center

9:00am-5:00pm

1 Hour Lunch Break

This four day program is ideal for those who are looking for an in-depth, hands-on introduction to Flow Cytometry.  The instructors will focus on a broad spectrum of flow cytometric topics, discuss examples from various research applications, provide technical protocols for flow cytometric sample preparation, as well as data analysis, trouble shooting and experimental design.  Attendees will have hands-on experience on three different flow platforms (Becton Dickinson, Miltenyi and Bio-Rad) as well as exposure to four software analysis programs.

Flow Cytometry platforms:   BD Accuri C6   I   Miltenyi Biotec MACSQuant Analyzer   I   Bio-Rad S3e
Flow Cytometry data analysis software:   FlowJo   I   FlowLogic    I    Cyflogic   I  FCS Express 6

NOTE: This course will also be offered November 6-9, 2017. 

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Lecture and Hands-on Interactive Training
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Team taught by active researchers
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Comprehensive binder containing workshop material
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Space limited to 18 participants
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Registration Fee: $995

Course Director

Elke Bergmann-Leitner, Ph.D.
Chief, Flow Cytometry Center
USMMRP/WRAIR

Lectures
Basic Analytical Techniques, Principles, and Trouble Shooting; Principles of Cell Staining, Choosing the Right Experimental Controls, Instrument Setup (baseline and compensation); Phenotyping and Analyzing Cell Populations by Flow Cytometry – How to present and interpret flow cytometric data; Introduction to acquisition and analysis Software; Purification and Labeling of Antibodies for Flow Cytometric and Other Ap­plications; Flow Sorting: Critical Experimental Issues and Instrument Setup; Parameters; Functional Flow Cytometric Assays;

Laboratory:
Preparation of Samples for Multicolor Analysis Flow Sorting; Multicolor Analysis, Compensation and Flow Sorting; Hands-on Flow Cytometry Exercises Using Miltenyi MACSQuant Analyzer; Bio-Rad S3e flow sorter; BD Accuri C6;  Software training: Introduction to FlowJo Cytometry Analysis Software; Introduction to Cyflogic Cytometry Analysis Software; Flow Cytometry - Acquisition/Trouble Shooting/ Software; Introduction to FCS Express Cytometry Analysis Software

Attendees attend all lectures and labs listed. 

Flow Cytometric Applications

  • Lecture: Basic Analytical Techniques, Principles, and Trouble Shooting
  • Lecture: Principles of Cell Staining, Choosing the Right Controls, Instrument Setup
  • Laboratory: Preparation of Samples for Multicolor Analysis Flow Sorting

Full Class Lectures

  • Multicolor Flow Cytometry and Panel Development
  • Phenotyping and Analyzing Cell Populations by Flow Cytometry - How to Read Flow Data

Small Learning Groups

  • Refer to ROTATIONS tab for descriptions

FACS Small Learning Groups

BD Accuri C6


STATION 1
Introductory Flow Cytometry Laboratory
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. Each attendee will setup a flow cytometer and perform manual compensation.

Miltenyi MACSQuant


STATION 2
Advanced Flow Cytometry Laboratory
Staining of Human PBMC with 7-color Immunophenotyping Flow Panel. Flow Cytometer Setup (MACSQuant; three lasers) with Special Emphasis on Compensation of Complex Multi-color Panels and Troubleshooting; Antibody Titrations as Preparative Tool for Panel Development; Utilization of Instrument for Rare Cell Detection. This Rotation Includes Automation Demonstrations Including Automated Compensation, Sample Labeling, Acquisition from a Multi-sampler and Analysis. In Addition to Immunophenotyping, Flow Cytometric Cell Counting and Nanoparticle Analysis will be Demonstrated.

Bio-Rad S3e Flow Sorter


STATION 3
Applied Laboratory
Flow Sorting of Peripheral Blood Mononuclear Cells (human) for Two Populations; Special 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.

Lecture

  • Analysis of cell cycle, cellular proliferation by flow cytometry

Laboratories

  • Data Workshop; Hands-on Data Analysis; Samples From Cell Cycle Analysis ; Apoptosis; Multi-Color Surface Staining; Proliferation using Bio-Tracers
  • Software training: Introduction to FlowJo Cytometry Analysis Software; Introduction to FlowLogic Cytometry Analysis Software; Introduction to Cyflogic Cytometry Analysis Software; Introduction to FCS Express Cytometry Analysis Software

Small Group Sessions

  • Flow Cytometry - Acquisition/Trouble Shooting/Software
  • 20 minute time slot rotations - student choice

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.

WLeitnerWolfgang Leitner, Ph.D.

Dr. Wolfgang Leitner the Chief of the Innate Immunity Section at the National Institute of Allergy and Infectious Diseases (NIH). He has been teaching flow cytometry for 25 years. He previously worked at the National Cancer Institute where he developed various flow cytometric protocols for the evaluation of experimental cancer vaccines.

The Bioscience Education Center

Montgomery College
20200 Observation Drive
Germantown, MD 20876