Probing Metabolism and Viscosity of Cancer Cells using Fluorescence Lifetime Imaging Microscopy

10.3791/62708 ◽  
2021 ◽  
Author(s):  
Liubov Shimolina ◽  
Maria Lukina ◽  
Vladislav Shcheslavskiy ◽  
Vadim Elagin ◽  
Varvara Dudenkova ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals Cannabidiol Modulates Mitochondrial Redox and Dynamics in MCF7 Cancer Cells: A Study Using Fluorescence Lifetime Imaging Microscopy of NAD(P)H

2021 ◽  
Vol 8 ◽  
Author(s):  
Rhys Richard Mould ◽  
Stanley W. Botchway ◽  
James R. C. Parkinson ◽  
Elizabeth Louise Thomas ◽  
Geoffrey W Guy ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Fluorescence Lifetime ◽  
Mitochondrial Function ◽  
Mode Of Action ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Mitochondrial Morphology ◽  
Fluorescence Lifetime Imaging ◽  
Dependent Manner ◽  
Lifetime Imaging ◽  
Dose Dependent

The cannabinoid, cannabidiol (CBD), is part of the plant's natural defense system that when given to animals has many useful medicinal properties, including activity against cancer cells, modulation of the immune system, and efficacy in epilepsy. Although there is no consensus on its precise mode of action as it affects many cellular targets, CBD does appear to influence mitochondrial function. This would suggest that there is a cross-kingdom ability to modulate stress resistance systems that enhance homeostasis. As NAD(P)H autofluorescence can be used as both a metabolic sensor and mitochondrial imaging modality, we assessed the potential of this technique to study the in vitro effects of CBD using 2-photon excitation and fluorescence lifetime imaging microscopy (2P-FLIM) of NAD(P)H against more traditional markers of mitochondrial morphology and cellular stress in MCF7 breast cancer cells. 2P-FLIM analysis revealed that the addition of CBD induced a dose-dependent decrease in bound NAD(P)H, with 20 µM treatments significantly decreased the contribution of bound NAD(P)H by 14.6% relative to the control (p < 0.001). CBD also increased mitochondrial concentrations of reactive oxygen species (ROS) (160 ± 53 vs. 97.6 ± 4.8%, 20 µM CBD vs. control, respectively, p < 0.001) and Ca2+ (187 ± 78 vs. 105 ± 10%, 20 µM CBD vs. the control, respectively, p < 0.001); this was associated with a significantly decreased mitochondrial branch length and increased fission. These are all suggestive of mitochondrial stress. Our results support the use of NAD(P)H autofluorescence as an investigative tool and provide further evidence that CBD can modulate mitochondrial function and morphology in a dose-dependent manner, with clear evidence of it inducing oxidative stress at higher concentrations. This continues to support emerging data in the literature and may provide further insight into its overall mode of action, not only in cancer, but potentially its function in the plant and why it can act as a medicine.

scholarly journals Mapping cisplatin-induced viscosity alterations in cancer cells using molecular rotor and fluorescence lifetime imaging microscopy

2020 ◽  
Vol 25 (12) ◽  
Author(s):  
Liubov E. Shimolina ◽  
Alexander A. Gulin ◽  
Miguel Paez-Perez ◽  
Ismael López-Duarte ◽  
Irina N. Druzhkova ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Molecular Rotor ◽  
Lifetime Imaging

scholarly journals Differential toxicity of gold-doxorubicin in cancer cells vs. cardiomyocytes as measured by real-time growth assays and fluorescence lifetime imaging microscopy (FLIM)

The Analyst ◽  
2015 ◽  
Vol 140 (16) ◽  
pp. 5732-5741 ◽  
Author(s):  
Eric Tawagi ◽  
Charlotte Massmann ◽  
Hicham Chibli ◽  
Jay L. Nadeau
Keyword(s):  
Cell Growth ◽  
Cancer Cells ◽  
Real Time ◽  
Gold Nanoparticle ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Differential Toxicity ◽  
Kinetics Of

The kinetics of toxicity of doxorubicin (Dox) and gold nanoparticle-conjugated doxorubicin (Au-Dox) were investigated in cultured B16 melanoma cells and cardiomyocytes using real-time cell-growth imaging.

scholarly journals Binding of Small Molecules to G-quadruplex DNA in Cells Revealed by Fluorescence Lifetime Imaging Microscopy of o-BMVC Foci

Molecules ◽  
2018 ◽  
Vol 24 (1) ◽  
pp. 35 ◽  
Author(s):  
Ting-Yuan Tseng ◽  
I-Te Chu ◽  
Shang-Jyun Lin ◽  
Jie Li ◽  
Ta-Chau Chang
Keyword(s):  
Cancer Cells ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Dna Integrity ◽  
Fluorescence Lifetime Imaging ◽  
Quadruplex Dna ◽  
Lifetime Imaging ◽  
G Quadruplex ◽  
Direct Binding ◽  
In Cells

G-quadruplex (G4) structures have recently received increasing attention as a potential target for cancer research. We used time-gated fluorescence lifetime imaging microscopy (FLIM) with a G4 fluorescent probe, 3,6-bis(1-methyl-2-vinylpyridinium) carbazole diiodide (o-BMVC), to measure the number of o-BMVC foci, which may represent G4 foci, in cells as a common signature to distinguish cancer cells from normal cells. Here, the decrease in the number of o-BMVC foci in the pretreatment of cancer cells with TMPyP4, BRACO-19 and BMVC4 suggested that they directly bind to G4s in cells. In contrast, the increase in the number of o-BMVC foci in the pretreatment of cells with PDS and Hoechst 33258 (H33258) suggested that they do not inhabit the binding site of o-BMVC to G4s in cells. After the H33258 was removed, the gradual decrease of H33258-induced G4 foci may be due to DNA repair. The purpose of this work is to introduce o-BMVC foci as an indicator not only to verify the direct binding of potential G4 ligands to G4 structures but also to examine the possible effect of some DNA binding ligands on DNA integrity by monitoring the number of G4 foci in cells.

scholarly journals Direct organelle thermometry with fluorescence lifetime imaging microscopy in single myotubes

2016 ◽  
Vol 52 (24) ◽  
pp. 4458-4461 ◽  
Author(s):  
Hideki Itoh ◽  
Satoshi Arai ◽  
Thankiah Sudhaharan ◽  
Sung-Chan Lee ◽  
Young-Tae Chang ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Heat Production ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
C2c12 Myotube

FLIM of ER thermo yellow and non-targeted mCherry reveals the Ca2+-dependent heat production localized to SR in C2C12 myotube.

Sign in / Sign up

Export Citation Format

Share Document