Introductory Course in Flow Cytometry Bio 632 March 6th – 8th 2018 – Cytometry Facility UZH

March 6th

9:15 – 13:00 Theory: Introduction to Flow Cytometry Y23G4

- 9:15 – 9:30 Welcome and course schedule overview - 9:30 – 10:15 Basics of Flow Cytometry - 10:30 – 11:15 Fluorochromes - 11:30 – 12:15 Applications for Flow Cytometry - 12: 30 – 13:00 Introduction to Diva Software incl CST

13:00 – 14:00 Lunch break

14:00 – 17:00 Practical Part Y44G03 => Tutors

- 14:00 – 14:45 Instrument start-up and quality control with CST beads - 14:45 – 15:30 Stain antibody titration with PBMCs and L/D control for PBMCs - 15:45 – 16:30 Acquisition of L/D PBMCs, Antibody titration, 8-Peak beads - 16:30 – 17:00 Data export

17:00 – 17:15 Wrap up and Questions with the tutors in front of the instruments

March 7th

9:15 – 12:00 Theory: Multicolor Flow Cytometry Y 23 G4

- 9:15 – 10:00 Controls in flow cytometry - 10:15 – 10:45 Spectral overlap and Compensation - 11:00 – 11:45 Experiment and panel design - 12:00 – 13:00 Workshop on panel design

o Guided lecture style, tutors to help with spectral viewer and instrument layout tables: Claudia, Andrea, Stephan

13:00 – 14:00 Lunch

14:00 – 17:00 Practical Part Y44G03 => Tutors

- 14:00 – 15:00 Stain 6 color T-reg Panel, FMO for CD25 and comp beads - 15:00 – 15:15 Instrument startup - 15:30 – 16:45 Acquisition Comp beads and 6 colour Treg Panel

incl. Compensation with Diva Software - 16:45 – 17:00 Data export

17:00 – 17:15 Wrap up and Questions with the tutors in front of the instruments

March 8th

9:00 – 12:30 Theory: Data Analysis either Y 21 D 68a

9:00 – 10:15 Manual Data analysis and visualization 10:20 – 10:40 Troubleshooting data before analysis 10:45 – 12:30 Data analysis with FlowJo

12:30-13:15 Lunch break 13:15– 16:15 Practical Part Y 11 J 05 - Hands on data analysis with FlowJo (SI calculation / Test Data sets / Treg Panel) 16:15 – 16:45 Exam

16:45 – 17:00 Final wrap up and Questions and Evaluation