Workshop on Flow Cytometry Experimental Designing and Data Analysis
11-12 March, 2015
Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), Bangalore
Fluorescent Activated Cell Sorting (FACS) or Flow Cytometry is widely used in basic and clinical research. To harness the real power of this state of art technique, appropriate experimental designing and accurate analysis of FACS data are crucial. Correct data analysis is not only important but also critical to adequately identify or functionally characterize the multiple populations of interest within the system. In the last few years, the number of parameters (colors) simultaneously used in flow cytometry experiments has increased. This is enabled by the availability of high-end instruments and powerful data analysis tools.
In this workshop, we will cover analysis of single & multicolor flow cytometry data and guidelines for the correct data analysis. Participants will be able to open their FCS files with the offline data analysis software, explore their data using various display options, perform gating, and quickly generate graphical and statistical reports to identify various populations of interest. Potential technical issues with the data will also be discussed. Study material will be provided that will include lectures, published articles and Purdue cytometry list discussions.
Date: 11-12 March, 2015
Venue: JNCSAR, Bangalore
Eligibility: Graduate students, postdoctoral researchers and other researchers from academic and non-academic institutions with prior experience in flow cytometry are invited for this course.
Registration: The tuition fee for the course is Rs. 3,500 for academic and Rs. 6500 for nonacademic participants. Pay in cheque/ DD/online transfer
How to apply: Those who are interested in attending the course may send in the following documents to; swaroopayalla@gmail.com / flowsols@gmail.com
1. Resume
2. Previous experience with flow cytometry
3. Reason for taking this course
4. Contact details
2. Previous experience with flow cytometry
3. Reason for taking this course
4. Contact details
Last date of application: 02 March, 2015
Participants will be notified of their selection by 6 th March, 2015.
Travel/ accommodation: Guest house is available for outstation candidates on paid basis.
Resource persons:
Dr. Hemant Agrawal, Instructor, Director,
Flowcytometry Solutions Pvt. Ltd.
Flowcytometry Solutions Pvt. Ltd.
Prof. Uday Kumar Ranga, HIV-AIDS Lab,
JNCASR, Bangalore
JNCASR, Bangalore
Contact:
Ms. Swaroopa Yalla
Flow Facility Incharge, JNCASR, Bangalore
Ph. No: +91-8494806079, +91-9176249644
E-mail ID: swaroopayalla@gmail.com
PROGRAMME OUTLINE
What is flow cytometry and its application?
Introduction to flow cytometry and some of the many applications of flow cytometry
How to design multicolour flow cytometry experiment?
How to design multicolour flow cytometry experiment?
An overview of experimental planning and optimization How to perform right compensation and its importance in data analysis?
Detailed overview of compensation and its importance in data analysis. How to make sense of flow cytometry data?
An overview of correct data analysis
How to report and present flow cytometry data?
Discussion about fundamental statistics and introduction to MiFlowCyte: Data presentation guidelines
How to analyze various data sets?
Demonstration and practice of data analysis for Immunophenotyping, Cell Cycle, Proliferation, Apoptosis, Viability Assay, Population comparison etc
How to troubleshoot flow cytometry data?
Guidelines to recognize and overcome bad data generation www.jncasr.ac.in www.flowsols.com
PROGRAMME DAY 1
9:30 - 10:30: Introduction to Flow cytometry and its applications
10:30 - 11:30: Designing of a Multicolor Flow Cytometry Experiment
- How to avoid Garbage In/Garbage Out Phenomenon
- Know your instrument.
- Selection of colors (Panel design)/selection of right antibodies
- Importance of dead cell marker.
- Experimental controls, Isotype controls, FMO, Biological controls, Internal controls
- Introduction to OMIPs—Orchestrating Multiplexity in Polychromatic Sciences.
11:30 - 11:45: Coffee/Tea Break
11:45 - 13:15: Compensation and Data Visualization
- Importance of right Compensation and how to perform it.
- Visualizing Data and Identifying Populations
13:15 - 14:00: Lunch
14:00 – 15:30: Basics of Flow Data Analysis and Presentation—What to do and what not to do?
15:30 – 15:45: Coffee/Tea break
15:45 - 17:30: Analysis of Flow Data and Reporting Results
10:30 - 11:30: Designing of a Multicolor Flow Cytometry Experiment
- How to avoid Garbage In/Garbage Out Phenomenon
- Know your instrument.
- Selection of colors (Panel design)/selection of right antibodies
- Importance of dead cell marker.
- Experimental controls, Isotype controls, FMO, Biological controls, Internal controls
- Introduction to OMIPs—Orchestrating Multiplexity in Polychromatic Sciences.
11:30 - 11:45: Coffee/Tea Break
11:45 - 13:15: Compensation and Data Visualization
- Importance of right Compensation and how to perform it.
- Visualizing Data and Identifying Populations
13:15 - 14:00: Lunch
14:00 – 15:30: Basics of Flow Data Analysis and Presentation—What to do and what not to do?
15:30 – 15:45: Coffee/Tea break
15:45 - 17:30: Analysis of Flow Data and Reporting Results
- Discussion about Fundamental Statistics
- Introduction to MiFlowCyte: Data Presentation Guidelines
- Introduction to MiFlowCyte: Data Presentation Guidelines
DAY 2
9:30 - 10:00: Review of the Day 1
10:00 - 11:30: Demonstration of flow data analysis—Cell Cycle, Proliferation, Apoptosis, Population comparison and multicolor assay.
11:30 - 11:45: Coffee/Tea Break
11:45 - 13:00: Troubleshooting flow cytometry data
13:00 - 14:00: Lunch
14:00 - 15:30: Hands on Data Analysis and Discussion
15:30 – 15:45: Coffee/Tea break
15:45 – 16:45: Data analysis and Discussion (continued)
16:45 - 17:30: Test and Feedback and Valedictory
9:30 - 10:00: Review of the Day 1
10:00 - 11:30: Demonstration of flow data analysis—Cell Cycle, Proliferation, Apoptosis, Population comparison and multicolor assay.
11:30 - 11:45: Coffee/Tea Break
11:45 - 13:00: Troubleshooting flow cytometry data
13:00 - 14:00: Lunch
14:00 - 15:30: Hands on Data Analysis and Discussion
15:30 – 15:45: Coffee/Tea break
15:45 – 16:45: Data analysis and Discussion (continued)
16:45 - 17:30: Test and Feedback and Valedictory
Participants should bring their own laptop and data files (.fcs files) for analysis and discussion. Data files will also be provided by us.
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