Imaging of neutral lipids and neutral lipid associated proteins

Methods Cell Biol. 2013:116:213-26. doi: 10.1016/B978-0-12-408051-5.00011-5.

Abstract

Intracellular fat droplets are large and have a distinct morphology, which makes their imaging at the light level simple and informative. We detail how to image the fat droplet core by metabolic labeling with fluorescent fatty acids or lipophilic fluorochromes. Further, we describe the use of indirect immunostaining to image fat droplet proteins and fat cores in the same field. We also address the use of appropriate controls for determining signal specificity and other practical considerations for optimizing image quality.

Keywords: BODIPY; Immunofluorescence; Immunostaining; Lipid droplets.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cytoplasm / metabolism
  • Cytoplasm / ultrastructure
  • Fatty Acids / metabolism*
  • Fluorescent Dyes / metabolism
  • Inclusion Bodies / chemistry
  • Inclusion Bodies / metabolism
  • Inclusion Bodies / ultrastructure*
  • Lipid Metabolism*
  • Lipids
  • Membrane Proteins / metabolism*
  • Membrane Proteins / ultrastructure
  • Microscopy, Fluorescence
  • Phosphoproteins / metabolism
  • Phosphoproteins / ultrastructure

Substances

  • Fatty Acids
  • Fluorescent Dyes
  • Lipids
  • Membrane Proteins
  • Phosphoproteins