Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy

Nan, Xiaolin; Cheng, Ji; Xie, Xuejun

doi:10.1194/jlr.d300022-jlr200

Cited by 303 publications

(270 citation statements)

References 37 publications

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“…Functional information can be provided by optical separation of molecular autofluorescence signals using time-resolved (FLIM) [23][24][25][26][27][28][29][30], spectral-resolved approaches [31][32][33][34] or both in tandem [35][36][37]. A complementary NLO imaging technique is coherent anti-Stokes Raman scattering (CARS) microscopy [38][39][40], which provides high-resolution images of the symmetric and asymmetric CH-stretching vibration [41][42][43], i.e. the main component in lipids.…”

Section: Introductionmentioning

confidence: 99%

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

Cicchi

Vogler

Kapsokalyvas

et al. 2012

Journal of Biophotonics

171

132

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Second‐harmonic generation (SHG) microscopy is a fantastic tool for imaging collagen and probing its hierarchical organization from molecular scale up to tissue architectural level. In fact, SHG combines the advantages of a non‐linear microscopy approach with a coherent modality able to probe molecular organization. In this manuscript we review the physical concepts describing SHG from collagen, highlighting how this optical process allows to probe structures ranging from molecular sizes to tissue architecture, through image pattern analysis and scoring methods. Starting from the description of the most relevant approaches employing SHG polarization anisotropy and forward – backward SHG detection, we then focus on the most relevant methods for imaging and characterizing collagen organization in tissues through image pattern analysis methods, highlighting advantages and limitations of the methods applied to tissue imaging and to potential clinical applications. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Section: Introductionmentioning

confidence: 99%

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

Cicchi

Vogler

Kapsokalyvas

et al. 2012

Journal of Biophotonics

171

132

View full text Add to dashboard Cite

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“…Concentration differences of the target molecule result in intensity variations of the CARS signal, forming the molecule-specific contrast in the image. The prospects of the technique have been shown for single adipocytes (13), axons (14), and even components in skin tissue of living mice (15).…”

mentioning

confidence: 99%

Monitoring of lipid storage in Caenorhabditis elegans using coherent anti-Stokes Raman scattering (CARS) microscopy

Hellerer

Axäng

Brackmann

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

294

290

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Better understanding of the fundamental mechanisms behind metabolic diseases requires methods to monitor lipid stores on single-cell level in vivo. We have used Caenorhabditis elegans as a model organism to demonstrate the limitations of fluorescence microscopy for imaging of lipids compared with coherent antiStokes Raman scattering (CARS) microscopy, the latter allowing chemically specific and label-free imaging in living organisms. CARS microscopy was used to quantitatively monitor the impact of genetic variations in metabolic pathways on lipid storage in 60 specimens of C. elegans. We found that the feeding-defective mutant pha-3 contained a lipid volume fraction one-third of that found in control worms. In contrast, mutants (daf-2, daf-4 dauer) with deficiencies in the insulin and transforming growth factors (IGF and TGF-␤) signaling pathways had lipid volume fractions that were 1.4 and 2 times larger than controls, respectively. This was observed as an accumulation of small-sized lipid droplets in the hypodermal cells, hosting as much as 40% of the total lipid volume in contrast to the 9% for the wild-type larvae. Spectral CARS microscopy measurements indicated that this is accompanied by a shift in the ordering of the lipids from gel to liquid phase. We conclude that the degree of hypodermal lipid storage and the lipid phase can be used as a marker of lipid metabolism shift. This study shows that CARS microscopy has the potential to become a sensitive and important tool for studies of lipid storage mechanisms, improving our understanding of phenomena underlying metabolic disorders.lipid metabolism ͉ nonlinear microscopy ͉ obesity

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“…Typically, Raman spectroscopy is used to identify different chemicals, such as lipids (Nan et al 2003), proteins (Uzunbajakava et al 2003) and myelin (Wang et al 2005), found in tissue.…”

Section: Raman Spectroscopymentioning

confidence: 99%

Novel imaging tools for investigating the role of immune signalling in the brain

Jacobsen

Parker

Everest‐Dass

et al. 2016

Brain, Behavior, and Immunity

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In all cases accepted manuscripts should:  link to the formal publication via its DOI  bear a CC-BY-NC-ND license -this is easy to do, click here to find out how  if aggregated with other manuscripts, for example in a repository or other site, be shared in alignment with our hosting policy  not be added to or enhanced in any way to appear more like, or to substitute for, the published journal article AbstractThe importance of neuro-immune interactions in both physiological and pathophysiological states cannot be understated. As our appreciation for the neuroimmune nature of the brain and spinal cord grows, so does our need to extend the spatial and temporal resolution of our molecular analysis techniques. Current imaging technologies applied to investigate the actions of the neuroimmune system in both health and disease states have been adapted from the fields of immunology and neuroscience and are inherently limited by their semiquantitative nature, loss of spatial resolution, photobleaching and detection sensitivity. Thus, the development of innovative methods which overcome these limitations are crucial for imaging and quantifying acute and chronic neuroimmune responses. Therefore, this review aims to convey novel and complementary imaging technologies in a form accessible to medical scientists engaging in neuroimmune research.

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Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy

Cited by 303 publications

References 37 publications

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

Monitoring of lipid storage in Caenorhabditis elegans using coherent anti-Stokes Raman scattering (CARS) microscopy

Novel imaging tools for investigating the role of immune signalling in the brain

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