UCD Flow Cytometry Summer School 2015

Transcription

1 UCD Flow Cytometry Summer School 2015 Flow Cytometry: Principles & Practice Monday May 25 th Day 1. Introduction to flow cytometry: principles and data analysis The main objective for day one is to be comfortable with the principles and basic concepts of flow cytometry and fluorescence; how to display and analyse the data and become familiar with standard operation procedures such as instrumentation basics, cleaning and maintenance, routine QC, and health and safety. During practical sessions, instruments and software from most manufacturers will be at hand, and basic operation procedures will be performed. 9:30-9:40 9:40-10:20 10:20-11:10 11:10-11:30 11:30-12:30 12:30-13:30 13:30-14:00 14: :45-15:00 15:00-15:45 15:45-16:15 16:15-16:45 Introduction to the Course. Alfonso Blanco Fluorochromes and Antibodies. Christopher Bullion Hardware. Alfonso Blanco Coffee (Sponsored by Jackson ImmunoResearch Europe) Data analysis & Statistics. Alfonso Blanco Lunch Practical session 1: Let s create an experiment. Optimising your Experiment I: Controls. Mark Cheetham Optimising your Experiment II: Voltages and Compensation. Barry Moran Practical session 2: Setting up an experiment. Practical session 3: 3 color experiment & compensation.

2 Tuesday May 26 th Day 2. FC instrumentation Day two entails a comprehensive approach to how flow cytometers and cell sorters work.the aim for today is to allow students to become familiar and more confident with how to run a flow cytometer (set voltages, gains, thresholds, etc.) based on the controls. We will explain which controls are needed, how to compensate correctly, selection of adequate reagents etc. In summary, to correctly set up a flow cytometry experiment. The hands on experience is to make participants comfortable with different instruments, showing them that once they know how to use one instrument they can conceivably run any flow cytometer. 9:30-10:15 10:15-11:00 11:00-11: :00-12:30 12:30-13:30 13:30-15:00 Cell Sorting. Mark Cheetham Tips and tricks for perfect labelling. Norman Maidment Coffee (Sponsored by Sony) Multi- colour Flow. Pannel Design. Norman Maidment Imaging Cytometry. Barry Lewis Lunch (Sponsored by Sony) Instrument Overview: Features and Benefits of the Latest Instrumentations. (Tour with Application Specialists) 13:30-13:40 Merck Millipore Guava 12HT Barry Lewis 13:40-13:50 Beckman Coulter CytoFlex Norman Maidment 13:50-14:00 Sony SH800Z Cell Sorter Roy Bongaerts 14:00-14:10 BioRad S3 cell sorter Mark Cheetham 14:10-15:00 How do they look? BL, NM, MC, BM, EB, AB 15:15-16:45 Practical Session. Instruments and remote sites. My first steps in flow. Where are my events?

3 Wednesday May 27 th Day 3. FC applications I: Apoptosis, cell proliferation and nanoparticle uptake For the first part of the application section, we will cover topics related to the biology of the cells; viability and cell proliferation. We will also introduce the participants to the nano world and how uptake may lead to proliferation changes and/or cytotoxicity. 9:30-10:30 10:30-11:00 11:00-11:20 11:20-12:10 12:10-12:40 12:40-13:30 13:30-15:00 15:45-16:45 Overview of Proliferation and Viability. Alfonso Blanco Nanoparticle Uptake and Cytotoxicity. Filippo Bertoli Coffee (Sponsored by BioStatus) From Drugs to DNA and Viability Markers. Roy Edward Molecular Characterisation and Regulation of Mitotic Catastrophe. Paul Lavin Lunch Practical Session I: Instruments and Remote Sites. Viability, cell cycle and nanoparticle uptake Practical Session II: Instruments and Remote Sites. Viability, cell cycle and nanoparticle uptake

4 Thursday May 28 th Day 4. FC applications II: Immunology and physiology of the cells During the second day of the application section participants will be introduced to immunophenotyping, intracellular signalling, cytokine staining, RNA indifferent cell types. We will also show the importance of considering time as one of the key parameters in this technology and their projects, leading to the introduction of Real Time Flow Cytometry. 9:30-10:20 10:20-11:00 11:00-11:15 11:15-12:00 12:00-12:40 12:40-13:30 13:30-15:00 15:15-16:45 Immunophenotyping. Achilleas Floudas Intracellular Cytokine Staining. Barry Moran Coffee Flowing small stuff. Jane Wood Time as a Parameter. Alfonso Blanco Lunch Practical Session I: Instruments and Remote Sites. Immunology, kinetics & small stuff Practical Session II: Instruments and Remote Sites. Immunology, kinetics & small stuff

5 Friday May 29 th Day 5. New Technologies. Participants at this point should understand the relevance of flow cytometry in the clinical field as well as research areas. Participants will be introduced to allied technologies such as mass spectrometry, multiplexing and other relatively new technologies that are filling the gap between flow cytometry and other platforms. As the final section, we will summarise the course, give some advice about the most common problems, how to analyse the data and present them following MIFlowCyt Standards. Finally, participants will have to perform an assessment (if required for ECTS). 9:30-10:15 10:15-10:45 10:45-11:00 11:00-11:30 11:30-12:30 12:00-14:00 14:00-14:30 14:30-15:00 15:15-16:15 16:15-16:30 16:30-17:00 Clinical Applications. Mark Cheetham Single Cell Genomics. Lisa Tang Coffee Analysis of extracellular signalling. Lisa Tang High Content Analysis. Jez Simpson Lunch Spectral Cytometry. Roy Bongaerts Overview of Single- Cell Technologies. Alfonso Blanco Troubleshooting, Resources & Publishing. Alfonso Blanco Conclusions. Alfonso Blanco Assessment

6 Speakers (in order of appearance) Alfonso Blanco Christopher Bullion Mark Cheetham Barry Moran Norman Maidment Barry Lewis Roy Bongaerts Filippo Bertoli Roy Edwards Paul Lavin Achilleas Floudas Jane Wood Lisa Tang Jeremy Simpson AbD Serotech BioRad Propel Labs Trinity College Dublin Beckman Coulter Merck Millipore Sony BioStatus Trinity College Dublin Sysmex MSC Instruments confirmed Amnis ImageStream BD Accuri C6 Beckman Coulter Cyan ADP Beckman Coulter Cyan ADP (modified by Propel Lab) Beckman Coulter CytoFLEX BioRadS3 Sony SH800Z Cell Sorter Sponsorship The organisers wish to thank sincerely all the companies participating in this event through provision of equipment, consumables and technical expertise. We are also grateful for additional sponsorship to facilitate networking opportunities during the course of the 2015 Flow Cytometry Summer School.