Flow Cytometry

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Hanson and I collected samples from 3hr and 6hr OCD-treated rat endothelial cells. These samples were separated into 12 test tubes and brought to the Core Facility in the Medical Science building for flow cytometry testing.

Flow cytometry is the diagnostic technique used to determine the cell count and their physical characteristics in a fluid medium.

Principle: Fluorescence is used in labelling cells. Fluorescent signal occurs when a molecule emits light from moving between a high excited state to an unexcited ground state. Similar to coimmunofluorescent staining, the flow cytometry allows for a comparative multi-parametic anaylsis using 2-4 compounds of different wavelengths.

FITC and PI are common fluorescent probes used.

Purpose: To distinguish the mode of death of the endothelial cells. Were they apoptotic or necrotic?

Results: We found that 30% of the cells underwent apoptotsis. These endothelial cells in the blood brain barrier may be subjected to programmed cell death in response to ischemic-repurfusion damage.