title: Fluorescence-Based In Situ Quantitative Imaging for Cellular Lipids.
abstract: Membrane lipids are dynamic molecules and their local concentrations serve as regulatory signals for diverse biological processes. To achieve quantitative in situ imaging of various lipids, we developed a ratiometric analysis using fluorescence biosensors, each of which is composed of an engineered lipid-binding protein and a covalently attached solvatochromic fluorophore. To cover a wide range of lipid concentration, lipid-binding proteins are engineered to have variable dynamic ranges. These tunable sensors allow robust and sensitive in situ quantitative lipid imaging in mammalian cells, providing new insight into the spatiotemporal dynamics and fluctuation of key signaling lipids. The sensor strategy is also applicable to in situ quantification of multiple cellular lipids or a single lipid in the opposing leaflets of cell membranes.
authors.affiliation: University of Illinois at Chicago, Chicago, IL, United States; Kyung Hee University, Yongin, South Korea. Electronic address: wcho@uic.edu.
authors.forename: W
authors.initials: W
authors.lastname: Cho
authors.affiliation: Konkuk University, Seoul, South Korea.
authors.forename: Y
authors.initials: Y
authors.lastname: Yoon
authors.affiliation: University of Illinois at Chicago, Chicago, IL, United States.
authors.forename: S-L
authors.initials: SL
authors.lastname: Liu
authors.affiliation: Kyung Hee University, Yongin, South Korea.
authors.forename: K
authors.initials: K
authors.lastname: Baek
authors.affiliation: University of Illinois at Chicago, Chicago, IL, United States.
authors.forename: R
authors.initials: R
authors.lastname: Sheng
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives / Q000737:chemistry; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003001:Cloning, Molecular; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000378:metabolism
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C475910:CLINT1 protein, human; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D019269:Phosphatidylinositol 4,5-Diphosphate; D011993:Recombinant Fusion Proteins; D053835:Unilamellar Liposomes; C038518:acrylodan; D015081:2-Naphthylamine; D005982:Glutathione Transferase; D003545:Cysteine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine
keywords: Fluorescence; Lipid imaging; Lipid sensors; Lipid signaling