Unit

UNIT 24.2 Fluorescent Detection of Lipid Droplets and Associated Proteins

  1. Laura L. Listenberger,
  2. Deborah A. Brown

Published Online: 1 JUN 2007

DOI: 10.1002/0471143030.cb2402s35

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Listenberger, L. L. and Brown, D. A. 2007. Fluorescent Detection of Lipid Droplets and Associated Proteins. Current Protocols in Cell Biology. 35:24.2:24.2.1–24.2.11.

Author Information

  1. State University of New York at Stony Brook, Stony Brook, New York

Publication History

  1. Published Online: 1 JUN 2007
  2. Published Print: JUN 2007

Abstract

Most eukaryotic cells can store excess lipid in cytosolic lipid droplets. This unit discusses techniques for the visualization of lipid droplets and associated proteins in cultured mammalian cells. Protocols for the detection of lipid droplets with nile red and BODIPY 493/503 are included. The differences in the spectral properties of these two lipophilic dyes and advantages of each are discussed. The best method for combining visualization of intracellular lipid droplets with indirect immunofluorescent detection of lipid droplet–associated proteins is described. Techniques for sample fixation and permeabilization must be chosen carefully to avoid alterations to lipid droplet morphology. Immunofluorescent detection of adipophilin, a broadly expressed, lipid droplet–associated protein, widely used as a marker for lipid droplet accumulation, is presented as an example. Finally, a simple protocol for enhancing lipid droplet accumulation through supplementation with excess fatty acid is included.

Keywords:

  • lipid droplets;
  • nile red;
  • BODIPY 493/503;
  • adipophilin;
  • PAT proteins