Raman imaging and lipidomic analysis of lipid droplets in (activated) hepatic stellate cells

Vaandrager, Arie B.; Testerink, N.; Ajat, Mokrish; Houweling, Martin; Brouwers, Jos F.; Pully, V.V.; Manen, H.J. van; Otto, Cornelis; Helms, J. Bernd

doi:10.1016/j.chemphyslip.2009.06.109

Cited by 1 publication

(1 citation statement)

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“…These approaches use the intrinsic vibrations of molecules for contrast and allow generation of chemically specific images with diffraction-limited optical resolution (Puppels et al 1990). Spontaneous raman microscopy is a well-established vibrational (hyperspectral) microscopy techniquesimilar to imaging mass spectrometry-and has been extensively used to study the local chemical composition of lDs in cultured cells (Matthaus et al 2012;Vaandrager et al 2009). However, the application of hyperspectral raman microscopy to study fresh tissue sections has been limited by two principle challenges: (1) the large autofluorescence response of tissues (Wang et al 2012) and (2) the limited penetration depth of the visible excitation wavelengths (Centonze and White 1998).…”

Section: Introductionmentioning

confidence: 99%

Chemical imaging of lipid droplets in muscle tissues using hyperspectral coherent Raman microscopy

Billecke

Rago

Bosma

et al. 2013

Histochem Cell Biol

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The accumulation of lipids in non-adipose tissues is attracting increasing attention due to its correlation with obesity. In muscle tissue, ectopic deposition of specific lipids is further correlated with pathogenic development of insulin resistance and type 2 diabetes. Most intramyocellular lipids are organized into lipid droplets (LDs), which are metabolically active organelles. In order to better understand the putative role of LDs in pathogenesis, insight into both the location of LDs and nearby chemistry of muscle tissue is very useful. Here, we demonstrate the use of label-free coherent anti-Stokes Raman scattering (CARS) microscopy in combination with multivariate, chemometric analysis to visualize intracellular lipid accumulations in ex vivo muscle tissue. Consistent with our previous results, hyperspectral CARS microscopy showed an increase in LDs in tissues where LD proteins were overexpressed, and further chemometric analysis showed additional features morphologically (and chemically) similar to mitochondria that colocalized with LDs. CARS imaging is shown to be a very useful method for label-free stratification of ectopic fat deposition and cellular organelles in fresh tissue sections with virtually no sample preparation.

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