1. Flow sorting – a brief overview Ian Titley Haematology Oncology Leukaemia Research Fund Institute of Cancer Research London

2. Flow sorting - a form of fluorescence microscopy whereby single cells in liquid suspension can be identified and physically separated from each other according to unique characteristics

3. History of cell sorter 1 st prototype at Los Alamos National Laboratory (LANL) in 1965 developed by Mack J Fulwyler by “joining” Coulter volume sensing (invented by Wallace and Joseph Coulter) with the newly invented ink-jet printer technology. Biologist Leonard Herzenberg at Stanford University was the first to recognise the utility for this technology to biological applications. With colleagues from the Genetics Dept Instrument Research group and using Fulwyler’s plans developed a machine to sort fluorescently labelled cells. The 1969 instrument had a mercury arc lamp as light source and in 1972 an argon ion laser. Funding from the National Institutes of Health (NIH) allowed Herzenberg and the Stanford engineers to interest the medical products company Becton Dickinson (BD) to convert their prototypes into the first commercial instruments, the FACS (Fluorescence Activated Cell Sorter) in 1975. Smithsonian Institute Archives – Video History Collection http://www.si.edu/archives/ihd/videocatalog/9554.htm

4. Electrostatic Mechanical “Lab-on-chip”

5. http://www.cardiff.ac.uk/medicine/haematology/cytonetuk/introduction_to_fcm/cell_sorting.htm

6. BD VantageSE DiVa option Beckman Coulter Epics Altra Hypersort DakoCytomation MoFloBD FACSAria

8. Sorting into a 96 well plate (Taken from BD FACSAria brochure)

10. Electrostatic ProsCons Fast (?x10 4 cellssec -1 )Expensive to buy and run MultiparameterNeeds trained operator High purityAerosol – biohazard 4-way

11. Electrostatic Mechanical “Lab-on-chip”

12. http://www.cardiff.ac.uk/medicine/haematology/cytonetuk/introduction_to_fcm/cell_sorting.htm

13. Union Biometrica COPAS BD FACSCalibur Partec PPCS

14. Mechanical ProsCons “Cheaper”Slow (?x10 2 cellssec -1 ) No aerosol1(2) way sorting Easier to use“Dilute” sorted sample Multiparameter

15. Electrostatic Mechanical “Lab-on-chip”

16. Gawad, S., Heuschkel, M., Leung-Ki, Y., Iuzzolino, R., Schild, L., Lerch, Ph. & Renaud, Ph.Fabrication of a microfluidic cell ananlyzer in a microchannel using impedance spectroscopy in IEEE-EMBS Conference on Microtechnologies in Medicine & Biology 297-301 (Lyon, France, 2000).

20. Flow cytometry: a practical approach MG Ormerod IRL Press Practical flow cytometry HM Shapiro WileyLiss Current protocols in flow cytometry Ed JP Robinson et al Wiley Useful texts

21. http://www.cardiff.ac.uk/medicine/haematology/cytonetuk/introduction_to_fcm/cell_sorting.htm Some cell sorting web resources http://www.rms.org.uk/cyto.shtml http://science.cancerresearchuk.org/sci/facs/fac_labinfo/flow_sorting/?version=1 http://www.cyto.purdue.edu/hmarchiv/index.htm Terry Hoy et al Cardiff University Royal Microscopical Society Derek Davies et al London Research Institute Cytometry E-mail archive Purdue USA