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Department of Pathology, University of Cambridge, UK

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Nigel Miller is the full-time operator for the Flow Cytometry facility in the Department of Pathology. Nigel has 27 years experience in flow cytometry. He provides a variety of services: on-hand troubleshooting and routine maintenance for all the machines and hands-on training for new users.

Nigel also runs a course in Flow cytometry for the Graduate School which includes theory and practical elements, technical advice on machine setup and data analysis, in addition he operates the MoFlo sorter.

Introduction to Flow Cytometry

Mondays at 9.25 a.m. in Pathology level 3

 Book through e mail   ngam2@cam.ac.uk 

               (same talk each week) commencing Oct 3rd 2011

If Mondays are inconvenient I can re-schedule to suit.

Talk - topics covered:-

  1. The cell. Light scattering parameters.
  2. Morphological and Genetic differences. Parasitology ?
  3. Lasers that produce enough power to visualize 100,000 cells/sec.
  4. Fluidics. Sheath fluid and sample are in co-axial flow.
  5. A typical analytical cytometer. FACScan,Cytek FACSCan and Cyan. Parameters, fluorochromes and filters*
  6. A typical sorting (MoFlo) cytometer. Setting up*
  7. Bone marrow. Identification of cell types. Stem cells. *
  8. Sample preparation. Mechanism of flow sorting. Compensation. *
  9. FSC,SSC and Propidium Iodide ( PI )-live/dead discrimination. *
  10. Cytokinesis. Relative merits of PI, Hoechst 33342,Sytox Green and DRAQ5  for DNA analysis. Plant and animal. *
  11. Cell surface (e.g. T and B lymphocytes) markers and isotypes controls.*
  12. Apoptosis. Learn to discriminate between Apoptosis,necrosis and Oncosis. AnnexinV, JC1,Hoechst and caspases. *
  13. The SP phenotype using Hoechst 33342. *
  14. Sorting x and y spermatozoa with Hoechst 33342.
  15. Chromosomes. Hoechst 33258 (AT) and Chromomycin A3 (GC)*
  16. Ca  ++ kinetics using Indo-1. *
  17. F.R.E.T. (fluorescence resonance electron transfer).
  18. Cell proliferation using C.F.S.E. *
  19. Pollen and associated starch grains
  20. From 20nm (Q dots) to 100u (2 cell mouse embryos).
  21. Publications, tutorials and useful websites
  22. RMS annual cyometry course.
  23. 23.The future -  The Amnis Imagestream /Accuri C6

*   Plus practical modules on  requested topics; e mail Nigel

 Nigel G A Miller, Flow Cytometrist

Immunology Division, Department of Pathology

University of Cambridge, Tennis Court Road,Cambridge,CB2 1QP, U.K.

e-mail: ngam2@cam.ac.uk; http://www.pathcam.ac.uk

http://www-immuno.path.cam.ac.uk/~immuno/flow/flowindex.html