Science courses not booked through the Graduate School

Flow cytometry training

Nigel Miller

Department of Pathology

Venue: Department of Pathology Level 3 (map)
Dates: 17th October 2011 onwards
Times: Mondays 9.25
Credits: 1

To book a place please e-mail Nigel Miller at ngam2@cam.ac.uk

An introductory talk on Flow Cytometry (a 'Cells in Flow' microscope) and cell sorting will be given to small groups. Each talk will last about 60-75 minutes and will be for a maximum of eight, each week.

Note that this talk is repeated each week. It is not one long course.

If Mondays are inconvenient it may be re-scheduled

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. Data analysis and statistics
10. FSC,SSC and Propidium Iodide ( PI )-live/dead discrimination *
11. Cytokinesis. Relative merits of PI, Hoechst 33342,Sytox Green and DRAQ5 for DNA analysis. Plant and animal *
12. Cell surface (e.g. T and B lymphocytes) markers and isotypes controls*
13. Apoptosis. Learn to discriminate between Apoptosis,necrosis and Oncosis. AnnexinV, JC1,Hoechst and caspases *
14. The SP phenotype using Hoechst 33342 *
15. Sorting x and y spermatozoa with Hoechst 33342
16. Chromosomes. Hoechst 33258 (AT) and Chromomycin A3 (GC)*
17. Ca++ kinetics using Indo-1 *
18. F.R.E.T. (fluorescence resonance electron transfer)
19. Cell proliferation using C.F.S.E. *
20. Pollen and associated starch grains
21. From 20nm (Q dots) to 100u (2 cell mouse embryos)
22. Publications, tutorials and useful websites
23. RMS annual cyometry course
24. The future - The Amnis Imagestream /Accuri C6

* Plus practical modules on requested topics: email Nigel