BuildingCYT_Flow Cytometry Training Courses @BI

After 11 years and following the training of over 2000 individual scientists from not just the UK but worldwide,  the highly successful Babraham Instiute Flow Cytometry Courses have been renewed and updated to include the most up to date information about this fast moving technology.  These new courses are now part of the 'BuildingCyt' which aims to building cytometry knowledge.

Flow Cytometry has been established in research and clinical laboratories for over 40 years and is a crucial technique in many investigations. However, the appreciation of how cytometers work and how best to set up and optimise an experiment is often lacking.

Established in 2014 these highly successful courses are designed by experts in flow cytometry and aimed at those relatively new to the field. The modular design of the courses allows delegates to build on knowledge from one course to the next, giving them the opportunity to implement ‘best practices’ and ultimately improving their flow cytometry experiments. It is recommended that delegates attend both modules.

The flow cytometry courses at BI are interactive, lecture and exercise based courses given by experts in the field who can share their in-depth knowledge of an evolving technology.   These courses are ideal for those using Flow Cytometry in Industry and Academia   These courses have been designed to build up knowledge of flow cytometry to ensure that the delegate is confident to design, carry out, analyse and present their flow cytometry data. 

Details of courses

Foundation Block: Introduction to Flow (24-27 delegates)

This one-day classroom-based course will introduce delegates to Flow Cytometry. We will cover the reagents used, how a cytometer works, experimental design, sample preparation and running, and data analysis and presentation. The course is aimed at those new to the field or those who want to increase their confidence in this technology. By the end of the course, delegates will be more confident in designing and running their own experiments.

Fluorescence and fluorochromes

How a Flow Cytometer works

Compensation

Experimental design

Sample preparation

Data analysis

Data presentation

Building Block: Spectral analysis by Flow Cytometry (15 delegates)

This one-day course will combine theory and practical. Delegates will learn what full spectrum cytometry can bring to their experiments. We will look at how full spectrum cytometers differ from conventional cytometers in their light detection, how this influences experimental design and some of the specific considerations of using this approach. We will run a simple experiment on two spectral cytometers - the Cytek Aurora and the Sony ID7000.

Fluorescence

Filters/lasers/detectors

Unmixing

Exp design

Machine Practicals – Aurora/ID7000

Advanced Flow Cytometry 20 delegates)

This course builds on the Introductory course with greater details on how a cytometer works, experimental design and cytometer optimization. As cytometers become capable of measuring an increasing number of fluorochromes, panel design becomes more complicated. It requires knowledge of the fluorochrome, the biology (antigen density, co-expression) and the sample preparation. In this one-day course we will look at the factors that need to be taken into account to build multicolour panels. We will do several interactive panel designs where we will use this knowledge to design panels of 6-10 and then 15+ colours.

Fluorochromes

Antigen density

Staining pattern

Fixation/Sample prep

Design considerations

Flow Cytometric Data analysis (18 delegates)

Data analysis of flow data files can be daunting. Where do I set regions and gates? Which metric do I use? In this one-day course we will guide delegates through the minefield of data analysis from simple gating to high dimensional analysis. This one-day course will introduce delegates to the analysis of flow data. In the first section, we will cover the plot types, setting compensation, regions and gates, the metrics that can be derived, and data presentation. The second section will look at the types of high dimensional analysis that are available, specifically dimensionality reduction and clustering methods. We will illustrate all these with data files that will be available to delegates.

Some sort of Intro

Which plots to use

Examples of gating

Metrics

Dim reduction and clustering

Hands on analysis using FlowJo and R

Cell sorting by flow cytometry (12 delegates)

Populations detected by analytical cytometry may be retrieved using droplet deflection cell sorting. Successful sorting requires knowledge of the sorting process and how to adjust parameters to maximise purity or yield There may be special considerations of downstream applications eg single cell sorting for RNAseq experiment. This two-day course will introduce delegates to the theory of cell sorting but will also include a practical element where we will use a cuvette-based system (FACSAria Fusion), a stream-in-air system (Thermo Bigfoot) and an automated system (Sony SH800).

Day One am: Refresher of how a cytometer works; nozzles; cuvette v stream in air; drop formation; drop delay

Day One pm: Practical – rotate between sorters– System walkthrough; setting drop delay/sorter set up

Day Two am: Sort masks; sample prep; other consideratons

Day Two pm: Sorting on different instruments

Practical Image Cytometry (18 delegates)

The ability to combine the phenotypic and quantitative information of a flow cytometer with the spatial and morphological insights of microscopy with imaging cytometry. This course goes through the need for imaging cytometry, how it works and practical considerations.

There are demonstrations within the BI flow facility using:

Imagestream MkII

FACSSDiscover S8

Attune CytPix

Testimonials from past courses:

'Great course leaders. Great course contents, great instructors Rachael and Derek delivered the course at the right speed, the workbook and exervice on panel design was very helpful to aid learning, thank you very much! Looking forward to the next course.'

'Great course that covers the basics and principles of flow cytometry for non-expert users'

'Clickers kept the course engaging. Leaders are certainly experts in their field. The learning involved in this course is logical, engaging and extremely clear!'

'Ideal to go over important theorertical topics. The positive vibe is really nice to keep us focussed'

For Terms and Conditions see http://www.babraham.ac.uk/science-services/flow-cytometry/flow-training