Automated quantitative analysis of lipid accumulation and hydrolysis in living macrophages with label-free imaging. 2013

Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 106, Taiwan.

The accumulation of lipids in macrophages is a key factor that promotes the formation of atherosclerotic lesions. Several methods such as biochemical assays and neutral lipid staining have been used for the detection of lipids in cells. However, a method for real-time quantitative assessment of the lipid content in living macrophages has yet to be shown, particularly for its kinetic process with drugs, due to the lack of suitable tools for non-invasive chemical detection. Here we demonstrate label-free real-time monitoring of lipid droplets (LDs) in living macrophages by using coherent anti-Stokes Raman scattering (CARS) microscopy. In addition, we have established an automated image analysis method based on maximum entropy thresholding (MET) to quantify the cellular lipid content. The result of CARS image analysis shows a good correlation (R(2) > 0.9) with the measurement of biochemical assay. Using this method, we monitored the processes of lipid accumulation and hydrolysis in macrophages. We further characterized the effect of a lipid hydrolysis inhibitor (diethylumbelliferyl phosphate, DEUP) and determined the kinetic parameters such as the inhibition constant, K(i). Our work demonstrates that the automated quantitative analysis method is useful for the studies of cellular lipid metabolism and has potential for preclinical high-throughput screening of therapeutic agents related to atherosclerosis and lipid-associated disorders.

UI MeSH Term Description Entries
D008055 Lipids A generic term for fats and lipoids, the alcohol-ether-soluble constituents of protoplasm, which are insoluble in water. They comprise the fats, fatty oils, essential oils, waxes, phospholipids, glycolipids, sulfolipids, aminolipids, chromolipids (lipochromes), and fatty acids. (Grant & Hackh's Chemical Dictionary, 5th ed) Lipid
D008264 Macrophages The relatively long-lived phagocytic cell of mammalian tissues that are derived from blood MONOCYTES. Main types are PERITONEAL MACROPHAGES; ALVEOLAR MACROPHAGES; HISTIOCYTES; KUPFFER CELLS of the liver; and OSTEOCLASTS. They may further differentiate within chronic inflammatory lesions to EPITHELIOID CELLS or may fuse to form FOREIGN BODY GIANT CELLS or LANGHANS GIANT CELLS. (from The Dictionary of Cell Biology, Lackie and Dow, 3rd ed.) Bone Marrow-Derived Macrophages,Monocyte-Derived Macrophages,Macrophage,Macrophages, Monocyte-Derived,Bone Marrow Derived Macrophages,Bone Marrow-Derived Macrophage,Macrophage, Bone Marrow-Derived,Macrophage, Monocyte-Derived,Macrophages, Bone Marrow-Derived,Macrophages, Monocyte Derived,Monocyte Derived Macrophages,Monocyte-Derived Macrophage
D008853 Microscopy The use of instrumentation and techniques for visualizing material and details that cannot be seen by the unaided eye. It is usually done by enlarging images, transmitted by light or electron beams, with optical or magnetic lenses that magnify the entire image field. With scanning microscopy, images are generated by collecting output from the specimen in a point-by-point fashion, on a magnified scale, as it is scanned by a narrow beam of light or electrons, a laser, a conductive probe, or a topographical probe. Compound Microscopy,Hand-Held Microscopy,Light Microscopy,Optical Microscopy,Simple Microscopy,Hand Held Microscopy,Microscopy, Compound,Microscopy, Hand-Held,Microscopy, Light,Microscopy, Optical,Microscopy, Simple
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D006868 Hydrolysis The process of cleaving a chemical compound by the addition of a molecule of water.
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013059 Spectrum Analysis, Raman Analysis of the intensity of Raman scattering of monochromatic light as a function of frequency of the scattered light. Raman Spectroscopy,Analysis, Raman Spectrum,Raman Optical Activity Spectroscopy,Raman Scattering,Raman Spectrum Analysis,Scattering, Raman,Spectroscopy, Raman
D050356 Lipid Metabolism Physiological processes in biosynthesis (anabolism) and degradation (catabolism) of LIPIDS. Metabolism, Lipid
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

Related Publications

Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
March 2010, Journal of lipid research,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
December 2017, Biomedical optics express,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
December 2008, Molecules and cells,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
October 2015, Proceedings of the National Academy of Sciences of the United States of America,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
November 2008, Biophysical journal,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
May 2017, Cytometry. Part A : the journal of the International Society for Analytical Cytology,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
November 2020, Analytical chemistry,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
January 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
July 2020, eLife,
Wei-Wen Chen, and Chen-Hao Chien, and Chiung-Lin Wang, and Huai-Hsien Wang, and Yuh-Lin Wang, and Shih-Torng Ding, and Tzong-Shyuan Lee, and Ta-Chau Chang
April 2013, Journal of proteome research,
Copied contents to your clipboard!