Phagocyte Biology Laboratory

Dr. Bryan Heit, Western University

In this short Youtube video we demonstrate a simple method of measuring cholesterol uptake and intracellular transport in macrophages. In this experiment, macrophages are cultured in a media containing oxLDL (bad cholesterol) and a cholesterol/lipid-reactive dye (Nile red). Live cell time-lapse microscopy is then used to monitor the accumulation of cholesterol by the cell. The movement of cholesterol within the cell can also be observed; accumulating first in the peri-nuclear region (within the Endoplasmic Reticulum) and later trafficking to discrete punctuate structures – lipid droplets. This is the first step in foam cell formation, with the stress of cholesterol accumulation eventually leading to the death of the cell.