Imaging of intracellular fatty acids by scanning X-ray fluorescence microscopy.
Fatty acids are taken up by cells and incorporated into complex lipids such as neutral lipids and glycerophospholipids. Glycerophospholipids are major constituents of cellular membranes. More than 1000 molecular species of glycerophospholipids differ in their polar head groups and fatty acid compositions. They are related to cellular functions and diseases and have been well analyzed by mass spectrometry. However, intracellular imaging of fatty acids and glycerophospholipids has not been successful due to insufficient resolution using conventional methods. Here, we developed a method for labeling fatty acids with bromine (Br) and applied scanning X-ray fluorescence microscopy (SXFM) to obtain intracellular Br mapping data with submicrometer resolution. Mass spectrometry showed that cells took up Br-labeled fatty acids and metabolized them mainly into glycerophospholipids in CHO cells. Most Br signals observed by SXFM were in the perinuclear region. Higher resolution revealed a spot-like distribution of Br in the cytoplasm. The current method enabled successful visualization of intracellular Br-labeled fatty acids. Single-element labeling combined with SXFM technology facilitates the intracellular imaging of fatty acids, which provides a new tool to determine dynamic changes in fatty acids and their derivatives at the single-cell level.-Shimura, M., Shindou, H., Szyrwiel, L., Tokuoka, S. M., Hamano, F., Matsuyama, S., Okamoto, M., Matsunaga, A., Kita, Y., Ishizaka, Y., Yamauchi, K., Kohmura, Y., Lobinski, R., Shimizu, I., Shimizu, T. Imaging of intracellular fatty acids by scanning X-ray fluorescence microscopy.
Department of Intractable Diseases National Center for Global Health and Medicine, Tokyo, Japan; mshimura@ri.ncgm.go.jp.|Riken SPring-8 Center, Hyogo, Japan.
Mari
M
Shimura
Lipid Signaling, Research Institute, National Center for Global Health and Medicine, Tokyo Japan; hsindou-tky@umin.net.|Agency for Medical Research and Development-Core Research for Evolutionary Science and Technology (AMED-CREST), Tokyo, Japan.
Hideo
H
Shindou
Riken SPring-8 Center, Hyogo, Japan.|Centre National de la Recherche Scientifique/University of Pau and Pays de l'Adour (CNRS/UPPA), Laboratoire de Chimie Analytique, Bio-Inorganique et Environnement (LCABIE), Unité Mixte de Recherche 5254, Pau, France.|Department of Chemistry of Drugs, Wroclaw Medical University, Wroclaw, Poland.
Lukasz
L
Szyrwiel
Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Suzumi M
SM
Tokuoka
Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.|Life Sciences Core Facility, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Fumie
F
Hamano
Department of Precision Science and Technology, Graduate School of Engineering, Osaka University, Osaka, Japan; and.
Satoshi
S
Matsuyama
Research Institute for Science and Engineering, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
Mayumi
M
Okamoto
Department of Intractable Diseases National Center for Global Health and Medicine, Tokyo, Japan.
Akihiro
A
Matsunaga
Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.|Life Sciences Core Facility, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Yoshihiro
Y
Kita
Department of Intractable Diseases National Center for Global Health and Medicine, Tokyo, Japan.
Yukihito
Y
Ishizaka
Department of Precision Science and Technology, Graduate School of Engineering, Osaka University, Osaka, Japan; and.
Kazuto
K
Yamauchi
Riken SPring-8 Center, Hyogo, Japan.
Yoshiki
Y
Kohmura
Centre National de la Recherche Scientifique/University of Pau and Pays de l'Adour (CNRS/UPPA), Laboratoire de Chimie Analytique, Bio-Inorganique et Environnement (LCABIE), Unité Mixte de Recherche 5254, Pau, France.
Ryszard
R
Lobinski
Research Institute for Science and Engineering, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
Isao
I
Shimizu
Lipid Signaling, Research Institute, National Center for Global Health and Medicine, Tokyo Japan.|Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Takao
T
Shimizu
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D020404:Glycerophospholipids / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids; D008856:Microscopy, Fluorescence / Q000379:methods; D014965:X-Rays
D005227:Fatty Acids; D020404:Glycerophospholipids; D008055:Lipids
cellular membrane; glycerophospholipid; labeling; lipid metabolism; visualization