Fluorescent lipid probes: some properties and applications (a review)

Maier, Olaf; Oberle, Volker; Hoekstra, Dick

doi:10.1016/s0009-3084(02)00017-8

Cited by 178 publications

(146 citation statements)

References 82 publications

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“…Finally, although these analogs resemble the properties of natural sphingolipids in many aspects (42,43), a direct comparison would be preferable. However, to specifically label SAC with sphingolipids or to accumulate newly synthesized sphingolipids in Golgi, the use of exchangeable or modifiable (dithionite quenching) sphingolipids is essential.…”

Section: Discussionmentioning

confidence: 99%

Trans-Golgi Network and Subapical Compartment of HepG2 Cells Display Different Properties in Sorting and Exiting of Sphingolipids

Maier¹,

Hoekstra

2003

Journal of Biological Chemistry

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In HepG2 cells, the subapical compartment (SAC) is involved in the biogenesis of membrane polarity. By contrast, direct apical transport originating from the trans-Golgi network (TGN), which may contribute to polarity establishment, has been poorly defined in these cells. Thus, although newly synthesized sphingolipids can be directly transported from the TGN to the apical membrane, numerous apical resident proteins are traveling via the transcytotic route. Here, we developed an in vitro transport assay and compared the molecular sorting of 6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino] hexanoyl-sphingomyelin (C 6 NBD-SM) and C 6 NBD-glucosylceramide (C 6 NBD-GlcCer) in TGN and SAC. SM is released from both TGN and SAC in the lumenal leaflet of transport vesicles. This holds also for GlcCer released from the SAC but not for a substantial fraction that departed from the Golgi. Distinct transport vesicles, enriched in either SM or GlcCer are released from SAC, consistent with their rigid sorting in this compartment. Different vesicle populations could not be recovered from TGN, although in situ experiments reveal that GlcCer is preferentially transported to the apical membrane, reflecting different transport mechanisms. The results indicate that in HepG2 cells sphingolipids are mainly sorted in the SAC membrane and that the release of SM from SAC and TGN is differentially regulated.

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Section: Discussionmentioning

confidence: 99%

Trans-Golgi Network and Subapical Compartment of HepG2 Cells Display Different Properties in Sorting and Exiting of Sphingolipids

Maier¹,

Hoekstra

2003

Journal of Biological Chemistry

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“…The fluorescent-dye diet method is biased by differences in food uptake efficiency, metabolic rate, distribution properties of the dye and the physical properties of the surrounding medium, as will be revealed in this study. In principle, the lipid molecules could also be specifically tagged by fluorescent marker molecules (5). However, the tags affect the intrinsic properties of the lipids due to their bulkiness and impact on the hydrophilic/hydrophobic balance (6), which in turn may change the cellular lipid trafficking and metabolism.…”

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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.

289

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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“…CARS imaging presents a unique opportunity to image lipids (and LDs), especially in live cells. Various studies have shown that measuring lipid quantities by applying well-known fluorescent stains such as Nile Red is unreliable as a consequence of non-specific lipid labelling, a problem that is alleviated with CARS imaging [19][20][21]. However, LipidTOX, a labelled phospholipid based stain, and Oil Red have been shown to be more reliable than the conventional Nile Red staining for neutral lipids [22].…”

Section: Lipid Droplet Generation and Apoptosismentioning

confidence: 99%

CARS based label‐free assay for assessment of drugs by monitoring lipid droplets in tumour cells

Steuwe

Patel

Ul-Hasan

et al. 2013

Journal of Biophotonics

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Coherent anti-Stokes Raman scattering (CARS) is becoming an established tool for label-free multi-photon imaging based on molecule specific vibrations in the sample. The technique has proven to be particularly useful for imaging lipids, which are abundant in cells and tissues, including cytoplasmic lipid droplets (LD), which are recognized as dynamic organelles involved in many cellular functions. The increase in the number of lipid droplets in cells undergoing cell proliferation is a common feature in many neoplastic processes [1] and an increase in LD number also appears to be an early marker of drug-induced cell stress and subsequent apoptosis [3]. In this paper, a CARS-based label-free method is presented to monitor the increase in LD content in HCT116 colon tumour cells treated with the chemotherapeutic drugs Etoposide, Camptothecin and the protein kinase inhibitor Staurosporine. Using CARS, LDs can easily be distinguished from other cell components without the application of fluorescent dyes and provides a label-free noninvasive drug screening assay that could be used not only with cells and tissues ex vivo but potentially also in vivo.Series of CARS images of HCT 116 cells after treatment at the specified times after drug treatment.

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Fluorescent lipid probes: some properties and applications (a review)

Cited by 178 publications

References 82 publications

Trans-Golgi Network and Subapical Compartment of HepG2 Cells Display Different Properties in Sorting and Exiting of Sphingolipids

Trans-Golgi Network and Subapical Compartment of HepG2 Cells Display Different Properties in Sorting and Exiting of Sphingolipids

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

CARS based label‐free assay for assessment of drugs by monitoring lipid droplets in tumour cells

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