Flow Cytometry: Colour Compensation

Transcription

1 Flow Cytometry: Colour Compensation (An intensive two day theoretical & demonstrated theories course) Course tutors: Ian Dimmick (Flow Cytometry Core Facility Manager) Owen Hughes (Flow Cytometry Specialist) Ian Harvey (Flow Cytometry Specialist) Newcastle University Flow Cytometry Core Facility Faculty of Medical Sciences, Bioscience Centre, International Centre for Life, Times Square, Newcastle Upon Tyne, NE1 3BZ, UK. Tel: Course date: 15 th & 16 th December 2010 The course is kindly sponsored by Becton Dickinson

2 Aim: The purpose of this course is to provide an understanding of the principles, concepts and technology of flow cytometry and the various functions that these technologies can fulfil in a clinical and research setting. Students will begin by getting a thorough theoretical grounding and then move on to supervised hands on/demonstrated experience of the instruments and experiments, including the setup of both basic and advanced multi coloured experiments. On completion of this course students will be able to interpret results from a wide range of experiments and variables, comprehend and manipulate the 3 main systems of flow cytometry (optics, electronics and fluidics) and be able to set up and run multicolour experiments on a broad range of flow cytometers with a high degree of confidence and competency. Group numbers are kept low (maximum of 8) to enable individual teaching and supervision, and to encourage discussion and problem solving of specific user applications. Students successfully completing this course will be awarded 10 University credits. Key topics: Instrument set up & quality control of instrumentation. Photodetector voltage settings (cells vs beads). The use of isotype controls and alternate strategies. The use of FMO controls. Instrument configurations. Selecting complimentary fluorochromes and dyes. Identifying spectral overlap and applying compensation. Hardware vs software compensation. Cells and/or beads as compensation controls. Compensation for non antibody based assays (e.g. GFP, viability dyes). The Dos and Don ts of multicolour compensation. Common problems and solutions. Sample experiments from 2 to 8 colour compensation. Questions.

3 Suggested reading: Hannon Fletcher Perry Maxwell; Advanced Techniques in Diagnostic cellular pathology; ISBN Hulspas R, O Gorman MRG, Wood GL, Gratama JW & Sutherland DR; Considerations for the control of background fluorescence in clinical flow cytometry; Cytometry Part B 2009; 76B: Dimmick I; The use of Qdot nanocrystals in flow cytometry: Novel reagents expand the scope of multicolour panels. Keeney M, Gratama JW, Chin Yee IH, Sutherland DR; Isotype Controls in the Analysis of Lymphocytes and CD34+ Stem and Progenitor Cells by Flow Cytometry Time to Let Go! BB36 417D 9ACE 6C77F6F6F229/27134/IsotypecontrolsTimetoletgo.pdf Sasaki DT, Maecker H, Trotter J; Establishing Optimum Baseline PMT Gains to Maximise Resolution on BD Biosciences Digital Flow Cytometers. Course dates: The next course will be held on 14th & 15th April 2010 (09:30am 17:00pm). Courses are run every three months. Following courses will be held in July 2010, October 2010 and January Course fee and registration: The registration fee for the two day course is 400 (lunch will be provided). To register for this course, please complete the registration form on the following page and post to Ian Dimmick at The Institute of Human Genetics, International Centre for Life, Central Parkway, Newcastle Upon Tyne, NE1 3BZ..

4 REGISTRATION DETAILS Name: Place of work: Tel: Address: Which instrumentation and software do you use? PAYMENT DETAILS Please e mail ian.dimmick@ncl.ac.uk and I will forward you a registration & payment form

5 Directions to venue: The Bioscience Centre is situated at the International Centre for Life, approximately 150m from Newcastle Central Station. Newcastle Upon Tyne International Airport is a 15 minute taxi ride, or 30 minutes using the tube. Newcastle University Flow Cytometry Core Facility, Faculty of Medical Sciences, Bioscience Centre, International Centre for Life, Times Square, Newcastle Upon Tyne, NE1 3BZ, UK. Recommended accommodation Jury's Inn The Holiday Inn The Station Hotel All are pleasant hotels and within 5 minutes walking distance from both the train station and the Bioscience Centre at the International Centre for Life.

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