Merck

Multiplex fatty acid imaging inside cells by Raman microscopy.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology (2020-06-28)
Masaaki Uematsu, Yoshihiro Kita, Takao Shimizu, Hideo Shindou
ABSTRACT

Visualizing intracellular fatty acids (including free and esterified form) is very useful for understanding how and where such molecules are incorporated, stored, and metabolized within cells. However, techniques of imaging multiple intracellular fatty acids have been limited by their small size, making it difficult to label and track without changing their biological and biophysical characteristics. Here, we present a new method for simultaneously visualizing up to five atomically labeled intracellular fatty acid species. For this, we utilized the distinctive Raman spectra depending on the labeling patterns and created a new, extensible opensource software to perform by-pixel analysis of extracting original spectra from mixed ones. Our multiplex imaging method revealed that fatty acids with more double bonds tend to concentrate more efficiently at lipid droplets. This novel approach contributes to reveal not only the spatial dynamics of fatty acids, but also of any other metabolites inside cells.

MATERIALS
Product Number
Brand
Product Description

Supelco
Supelco 37 Component FAME Mix, certified reference material, TraceCERT®, in dichloromethane (varied conc.), ampule of 1 mL
Sigma-Aldrich
PF-06424439, ≥98% (HPLC)
Sigma-Aldrich
T863, ≥98% (HPLC)