Flow cytometry is a powerful analytical tool that can analyze up to 10,000 individual cells per second. It works by passing single cells in suspension through a flow cell where they are exposed to a laser. Light scattering and fluorescence emissions from stained cells are then measured using detectors. Flow cytometry has many applications including analyzing cell viability, cell cycle, apoptosis, and cell sorting. It provides benefits like rapid analysis of single cells but also has limitations such as not showing intracellular protein localization and requiring cell preparation.

1. Flow Cytometry Anna Merca - Philippe Melas - Tolga Sütlü Stockholm Research School in Molecular Life Sciences 14 October 2005

5. Direct beam stop Laser Light High angle scatter : (Side Scatter) Cell structure Low angle scatter : (Forward Scatter) Cell size Fluorescence at longer wavelengths Flow cell Direction of flow

6. Basic Optics of a Flow Cytometer ( An automated fluorescent microscope) Dichroic mirrors 1 2 3 Laser(s) Cell Collection Lenses Scatter Low & High angle Photomultiplier tubes

8. Analyzing the graph -one color- CD3 NUMBER CD4 NUMBER CD8 NUMBER

9. Analyzing the graph CD4 0 1 10 2 10 3 10 4 0 10 CD3 CD3 CD4 CD3+CD4+ green CD3-CD4+ cyan CD3+CD4- cyan CD3-CD4- black

11. Apoptosis vs. Necrosis

16. Cell Sorting 2 Charged Plates Collection Tubes

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