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Flow cytometry training

Venue: Department of Pathology Level 3 (map)
Dates:7th October 2013 onwards
Times: Mondays 9.25
Credits: 1

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

Nigel Miller from the Department of Pathology runs short introductory talks on Flow Cytometry on Mondays throughout each term. The sessions are designed so that an individual only needs to attend the session once, but practical demonstrations and further training can be arranged on request.

If you are interested, please email ngam2@cam.ac.uk. Nigel tries to arrange the sessions so that researchers with common subject areas can attend the same session, rather than trying to cover all topics for all researchers each time. If you know a group of researchers with similar needs, then you may all be able to attend the same session.

Each talk will last about 60-75 minutes and will be for a maximum of eight, each week.

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