Lipid-cell interactions in human monocytes investigated by doubly-resonant coherent anti-Stokes Raman scattering microscopy

Weeks, Tyler; Schie, Iwan W.; Hartigh, Laura J. den; Rutledge, John C.; Huser, Thomas

doi:10.1117/1.3544585

Cited by 26 publications

(23 citation statements)

References 19 publications

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“…However, from the literature, monocytes were reported to uptake fatty acids from very low-density lipoprotein lipolysis products and formed small intracellular lipid droplets [44-46]. The lipid contents of monocytes were low overall and declined postprandially [45].…”

Section: Resultsmentioning

confidence: 99%

Detection of Lipid-Rich Prostate Circulating Tumour Cells with Coherent Anti-Stokes Raman Scattering Microscopy

et al. 2012

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BackgroundCirculating tumour cells (CTC) are an important indicator of metastasis and associated with a poor prognosis. Detection sensitivity and specificity of CTC in the peripheral blood of metastatic cancer patient remain a technical challenge.MethodsCoherent anti-Stokes Raman scattering (CARS) microscopy was employed to examine the lipid content of CTC isolated from the peripheral blood of metastatic prostate cancer patients. CARS microscopy was also employed to evaluate lipid uptake and mobilization kinetics of a metastatic human prostate cancer cell line.ResultsOne hundred CTC from eight metastatic prostate cancer patients exhibited strong CARS signal which arose from intracellular lipid. In contrast, leukocytes exhibited weak CARS signal which arose mostly from cellular membrane. On average, CARS signal intensity of prostate CTC was 7-fold higher than that of leukocytes (P<0.0000001). When incubated with human plasma, C4-2 metastatic human prostate cancer cells exhibited rapid lipid uptake kinetics and slow lipid mobilization kinetics. Higher expression of lipid transport proteins in C4-2 cells compared to non-transformed RWPE-1 and non-malignant BPH-1 prostate epithelial cells further indicated strong affinity for lipid of metastatic prostate cancer cells.ConclusionsIntracellular lipid could serve as a biomarker for prostate CTC which could be sensitively detected with CARS microscopy in a label-free manner. Strong affinity for lipid by metastatic prostate cancer cells could be used to improve detection sensitivity and therapeutic targeting of prostate CTC.

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Section: Resultsmentioning

confidence: 99%

Detection of Lipid-Rich Prostate Circulating Tumour Cells with Coherent Anti-Stokes Raman Scattering Microscopy

et al. 2012

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“…For example, van Manen et al used d 8 -arachidonic acid (AA) to label LDs in neutrophils, and observed close association between AA-enriched LDs and phagosome 41 . Xie et al 42 and Weeks et al 43 demonstrated that d 33 -oleic acid or d 2 -oleic acid could be used for CARS microscopic imaging of lipid droplet. With reduced non-resonant background in SRS, Zhang et al were able to reveal the cellular uptake of d 31 -palmitic acid and its incorporation into LDs and membrane 44 .…”

Section: Lipid Metabolismmentioning

confidence: 99%

Applications of vibrational tags in biological imaging by Raman microscopy

Zhao

Shen

et al. 2017

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As a superb tool to visualize and study the spatial-temporal distribution of chemicals, Raman microscopy has made big impacts to many disciplines of science. While the label-free imaging has been the prevailing strategy in Raman microscopy, recent development and applications of vibrational/Raman tags, particularly when coupled with stimulated Raman Scattering (SRS) microscopy, have generated intense excitement in biomedical imaging. SRS imaging of vibrational tags has enabled researchers to study a wide range of small biomolecules with high specificity, sensitivity and multiplex capability, at single live cell level, tissue level or even in vivo. As reviewed in the article, this platform has facilitated imaging distribution and dynamics of small molecules such as glucose, lipids, amino acids, nucleic acids, and drugs that are otherwise difficult to do with other means. As both the vibrational tags and Raman instrumental development progress rapidly and synergistically, we anticipate that the technique will shed light onto an even broader spectrum of biomedical problems.

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“…CARS microscopy has been extensively applied to the study and visualization of LDs in cells and in tissue. [14][15][16][17][18] In particular, CARS microscopy enabled for the rst time the rapid label-free visualization of cellular and tissue lipid accumulations with chemical specicity and single cell and single lipid droplet sensitivity. In our own work, we have recently shown that we can quantify the effects of lipolysis products from very low density lipoproteins on cells and determine the average degree of unsaturation in individual cellular lipid droplets based on the analysis of single Raman peak ratios.…”

Section: Introductionmentioning

confidence: 99%

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

Schie

Nolte

Pedersen

et al. 2013

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Cellular lipid droplets are the least studied and least understood cellular organelles in eukaryotic and prokaryotic cells. Despite a significant body of research studying the physiology of lipid droplets it has not yet been possible to fully determine the composition of individual cellular lipid droplets. In this paper we use Raman spectroscopy on single cellular lipid droplets and least-squares fitting of pure fatty acid spectra to determine the composition of individual lipid droplets in cells after treatment with different ratios of oleic and palmitic acid. We validate the results of the Raman spectroscopy-based single lipid droplet analysis with results obtained by gas chromatography analysis of millions of cells, and find that our approach can accurately predict the relative amount of a specific fatty acid in the lipid droplet. Based on these results we show that the fatty acid composition in individual lipid droplets is on average similar to that of all lipid droplets found in the sample. Furthermore, we expand this approach to the investigation of the lipid composition in single cellular peroxisomes. We determine the location of cellular peroxisomes based on two-photon excitation fluorescence (TPEF) imaging of peroxisomes labeled with the green fluorescent protein, and successive Raman spectroscopy of peroxisomes. We find that in some cases peroxisomes can produce a detectable CARS signal, and that the peroxisomal Raman spectra exhibit an oleic acid-like signature.

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Lipid-cell interactions in human monocytes investigated by doubly-resonant coherent anti-Stokes Raman scattering microscopy

Cited by 26 publications

References 19 publications

Detection of Lipid-Rich Prostate Circulating Tumour Cells with Coherent Anti-Stokes Raman Scattering Microscopy

Detection of Lipid-Rich Prostate Circulating Tumour Cells with Coherent Anti-Stokes Raman Scattering Microscopy

Applications of vibrational tags in biological imaging by Raman microscopy

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

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