Direct comparison of fatty acid ratios in single cellular lipid droplets as determined by comparative Raman spectroscopy and gas chromatography

Schie, Iwan W.; Nolte, Lena; Pedersen, Theresa L.; Smith, Zachary J.; Wu, Jian; Yahiatène, Idir; Newman, John W.; Huser, Thomas

doi:10.1039/c3an00970j

Cited by 56 publications

(51 citation statements)

References 32 publications

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“…In a separate study, we validated specific prediction of fatty acid profile of lipid droplets within hepatic cells by Raman spectroscopy with data from gas chromatographic analysis of fatty acid content. 26 The data of Raman spectral prediction are consistent with the data by mass spectral analysis of fatty acids. 20 These findings confirmed the notion that there is a higher ratio of saturated fatty acid over unsaturated fatty acids in lipid droplets.…”

Section: Discussionsupporting

confidence: 82%

A novel mouse model of nonalcoholic steatohepatitis with significant insulin resistance

Adkins

Schie

Fedor

et al. 2013

Laboratory Investigation

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Currently available models insufficiently reflect the pathogenic alternation of nonalcoholic steatohepatitis\NASH), such as insulin resistance. The present study aimed to characterize a novel NASH model caused by feeding the diet containing conjugated linoleic acid (CLA). In this study, mice were fed a control diet or the diet containing 0.5% CLA for 8 weeks. The insulin tolerance test (ITT) and homeostasis model assessment of insulin resistance (HOMA-IR) were used to determine the extent of insulin resistance. Liver lipotoxicity and inflammation were assessed by endoplasmic reticulum (ER) stress, autolipophagy, recruitment of Kupffer cells and hepatic stellate cell (HSC) activation. We found that liver weight was markedly increased, and histopathological examination showed marked macrosteatosis with focal hepatocellular death through apoptosis, and mild pericellular fibrosis with Kupffer cell recruitment and HSC activation, as well as light chain IIIb-positive cells and enhanced ER stress in mice fed the CLA-containing diet. Enhanced synthesis and reduced b-oxidation of fatty acids resulted in their accumulation and lipotoxicity in hepatocytes. A biophotonic technology revealed lipid droplet accumulation in the liver from mice fed the CLA-containing diet, and Raman spectroscopic analysis indicated that these lipid droplets predominantly contained saturated fatty acids. Elevated fasting insulin levels, abnormal ITT and HOMA-IR confirmed the marked insulin resistance in these mice. Decreased phosphorylation of the insulin-signaling molecule Akt was partially responsible for the significant insulin resistance. In conclusion, Mice fed the diet containing CLA-developed steatohepatitis with marked insulin resistance, which is similar to the characteristics observed in NASH patients. The further characterization of this model would be particularly useful for revealing the critical role of insulin resistance in NASH development in conditions such as metabolic syndrome, diabetes and obesity.

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Section: Discussionsupporting

confidence: 82%

A novel mouse model of nonalcoholic steatohepatitis with significant insulin resistance

Adkins

Schie

Fedor

et al. 2013

Laboratory Investigation

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“…In the present work we tested hypothesis that AA is up taken and stored in LDs and its uptake via cellular membrane may be regulated by cationic compounds. LDs formation upon PUFAs uptake has been demonstrated previously in various cells with the use of Raman spectroscopy but not in endothelium. We have recently shown for the first time that endothelial cells are able to form LDs in response to arachidonic acid uptake .…”

Section: Introductionmentioning

confidence: 99%

Lipid droplets formation in human endothelial cells in response to polyunsaturated fatty acids and 1‐methyl‐nicotinamide (MNA); confocal Raman imaging and fluorescence microscopy studies

Majzner

Chłopicki

Barańska

2015

Journal of Biophotonics

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In this work the formation of lipid droplets (LDs) in human endothelial cells culture in response to the uptake of polyunsaturated fatty acids (PUFAs) was studied. Additionally, an effect of 1-methylnicotinamide (MNA) on the process of LDs formation was investigated. LDs have been previously described structurally and to some degree biochemically, however neither the precise function of LDs nor the factors responsible for LD induction have been clarified. Lipid droplets, sometimes referred in the literature as lipid bodies are organelles known to regulate neutrophil, eosinophil, or tumor cell functions but their presence and function in the endothelium is largely unexplored. 3D linear Raman spectroscopy was used to study LDs formation in vitro in a single endothelial cell. The method provides information about distribution and size of LDs as well as their composition. The incubation of endothelial cells with various PUFAs resulted in formation of LDs. As a complementary method for LDs identification a fluorescence microscopy was applied. Fluorescence measurements confirmed the Raman results suggesting endothelial cells uptake of PUFAs and subsequent LDs formation in the cytoplasm of the endothelium. Furthermore, MNA seem to potentiate intracellular uptake of PUFAs to the endothelium that may bear physiological and pharmacological significance. Confocal Raman imaging of HAoEC cell with LDs.

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“…The spectra of these three macromolecules are shown in Figure B. To determine the relative macromolecule at a specific location of the sample, each measured spectrum was preprocessed and least‐squares fitted by the molecular signatures of interest from the database . To visualize the information each acquired spectrum during imaging procedure is assigned an RGB color, representing the relative contribution of the different chemical components.…”

Section: Methodsmentioning

confidence: 99%

Raman ChemLighter: Fiber optic Raman probe imaging in combination with augmented chemical reality

Yang

Mondol

Stiebing

et al. 2019

Journal of Biophotonics

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Raman spectroscopy using fiber optic probe combines non‐contacted and label‐free molecular fingerprinting with high mechanical flexibility for biomedical, clinical and industrial applications. Inherently, fiber optic Raman probes provide information from a single point only, and the acquisition of images is not straightforward. For many applications, it is highly crucial to determine the molecular distribution and provide imaging information of the sample. Here, we propose an approach for Raman imaging using a handheld fiber optic probe, which is built around computer vision–based assessment of positional information and simultaneous acquisition of spectroscopic information. By combining this implementation with real‐time data processing and analysis, it is possible to create not only fiber‐based Raman imaging but also an augmented chemical reality image of the molecular distribution of the sample surface in real‐time. We experimentally demonstrated that using our approach, it is possible to determine and to distinguish borders of different bimolecular compounds in a short time. Because the method can be transferred to other optical probes and other spectroscopic techniques, it is expected that the implementation will have a large impact for clinical, biomedical and industrial applications.

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Direct comparison of fatty acid ratios in single cellular lipid droplets as determined by comparative Raman spectroscopy and gas chromatography

Cited by 56 publications

References 32 publications

A novel mouse model of nonalcoholic steatohepatitis with significant insulin resistance

A novel mouse model of nonalcoholic steatohepatitis with significant insulin resistance

Lipid droplets formation in human endothelial cells in response to polyunsaturated fatty acids and 1‐methyl‐nicotinamide (MNA); confocal Raman imaging and fluorescence microscopy studies

Raman ChemLighter: Fiber optic Raman probe imaging in combination with augmented chemical reality

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