Single‐ and two‐photon spectral imaging of intrinsic fluorescence of transformed human hepatocytes

Rajwa, Bartek; Bernaś, Tytus; Acker, H.; Dobrucki, Jurek; Robinson, J. Paul

doi:10.1002/jemt.20497

Cited by 10 publications

(6 citation statements)

References 37 publications

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“…Autofluorescence of cells is documented and also used to determine metabolic state of cells, mainly hepatic cells [33,59]. The high fluorescent is reported to be due to the fluorescence properties of reduced pyridine nucleotide NAD(P)H (emitting in the range of 440 to 480 nm after UV-excitation) and flavins FAD/FMN (emitting in the range of 520 nm after 488 nmexcitation) as major intracellular fluorophores [32,34]. By the way, resveratrol and viniferin (resveratrol dimer) don't seem to have similar targets since only resveratrol and resveratrol triacetate induce green cells autofluorescence.…”

Section: Discussionmentioning

confidence: 99%

“…Flow cytometry analyses by UV excitation reveal that resveratrol related compounds induce an increase of cells autofluorescence. It is also known that reduced pyridine nucleotide NAD(P)H is the major intracellular fluorophore witch presents a 338 nm excitation maximum and emits in the range of 440 to 480 nm [32][33][34]. We also decided to quantify the consequences of resveratrol related compounds treatments on cellular NADPH content.…”

Section: Quantification Of Nadph Cell Content Induction By Resveratromentioning

confidence: 99%

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Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses

et al. 2008

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Resveratrol is a well known polyphenol largely produced in grapevine. It is a strong antioxidant and a free radical scavenger. It exhibits several beneficial effects for health including cancer. Resveratrol antioxidant activity is essential in the prevention of chemical-induced cancer by inhibiting initiation step of carcinogenesis process but it is also considered to inhibit cancer promotion and progression steps. While the effects of resveratrol on cancer cells are widely described, the data available on the antiproliferative potential of resveratrol derivatives remain weak. Nevertheless, resveratrol analogs could exhibit stronger potentials than the parent molecule. So, we compared the cellular effects of trans-resveratrol, trans-epsilon-viniferin and their respective acetate derivatives, as well as a polyphenol mixture extracted from grapevine shoots, called vineatrol. We studied their abilities to interfere with cell proliferation, their uptake and their effects on parameters of cellular state in human hepatoma cells (HepG2). Cell growth experiments show that resveratrol triacetate presents a slightly better antiproliferative potential than resveratrol. The dimer epsilon-viniferin,as well as its pentaacetate analog, is less powerful than resveratrol, although a similar uptake kinetics in cells. Interestingly, among the tested polyphenols, vineatrol is the most potent solution, indicating a possible synergistic effect of both resveratrol and epsilon-viniferin. We took advantage of the fluorescence properties of these compounds to evidence cellular uptake by using flow cytometry. In addition, by competition assay, we demonstrate that resveratrol triacetate enters in hepatic HepG2 cells by the same way as resveratrol. By autofluorescence in situ measurement we observed that resveratrol and related compounds induce deep changes in cells activity. These changes occur mainly by increasing NADPH cell content and the number of green fluorescent cytoplasmic granular structures which may be related to an induction of detoxifying enzyme mechanisms.

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Section: Discussionmentioning

confidence: 99%

Section: Quantification Of Nadph Cell Content Induction By Resveratromentioning

confidence: 99%

Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses

et al. 2008

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“…Our results also demonstrate the ability to discriminate, in vivo , between different fluorescent molecules and the possibility of acquiring diverse, complementary information such as quantitative/environmental data simultaneously [25,26]. …”

Section: Discussionmentioning

confidence: 85%

Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications

Hwang¹,

Wachsmann‐Hogiu²,

Ramanujan³

et al. 2011

Biomed. Opt. Express

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We report fast, non-scanning, wide-field two-photon fluorescence excitation with spectral and lifetime detection for in vivo biomedical applications. We determined the optical characteristics of the technique, developed a Gaussian flat-field correction method to reduce artifacts resulting from non-uniform excitation such that contrast is enhanced, and showed that it can be used for ex vivo and in vivo cellular-level imaging. Two applications were demonstrated: (i) ex vivo measurements of beta-amyloid plaques in retinas of transgenic mice, and (ii) in vivo imaging of sulfonated gallium(III) corroles injected into tumors. We demonstrate that wide-field two photon fluorescence excitation with flat-field correction provides more penetration depth as well as better contrast and axial resolution than the corresponding one-photon wide field excitation for the same dye. Importantly, when this technique is used together with spectral and fluorescence lifetime detection modules, it offers improved discrimination between fluorescence from molecules of interest and autofluorescence, with higher sensitivity and specificity for in vivo applications.

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“…This phenomenon is due to the oxidized forms of riboflavin, the co-enzymes flavin and NADH reduced inside cells 7,8. Other molecules like lipofuscin and ceroides or other components of the skin, such as collagen and melanin, also contribute to this effect.…”

Section: Fluorescence Imagingmentioning

confidence: 99%

Fluorescence imaging agents in cancerology

Paganin-Gioanni¹,

Bellard²,

Paquereau³

et al. 2010

Radiology and Oncology

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BackgroundOne of the major challenges in cancer therapy is to improve early detection and prevention using novel targeted cancer diagnostics. Detection requests specific recognition. Tumor markers have to be ideally present on the surface of cancer cells. Their targeting with ligands coupled to imaging agents make them visible/detectable.ConclusionsFluorescence imaging is a newly emerging technology which is becoming a complementary medical method for cancer diagnosis. It allows detection with a high spatio-temporal resolution of tumor markers in small animals and in clinical studies. In this review, we focus on the recent outcome of basic studies in the design of new approaches (probes and devices) used to detect tumor cells by fluorescence imaging.

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Single‐ and two‐photon spectral imaging of intrinsic fluorescence of transformed human hepatocytes

Cited by 10 publications

References 37 publications

Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses

Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses

Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications

Fluorescence imaging agents in cancerology

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