Targetable and fixable rotor for quantifying mitochondrial viscosity of living cells by fluorescence lifetime imaging

Song, Xinbo; Li, Ning; Wang, Chao; Xiao, Yi

doi:10.1039/c6tb02524b

Cited by 92 publications

(49 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is important to monitor the mitochondrial membrane potential. Carbonyl cyanide 3‐chlorophenylhydrazone (CCCP) can induce the decrease of the mitochondria membrane potential rapidly 30–33. Thus, the response of the MMP‐CDs toward the CCCP induced mitochondrial membrane potential decrease was then examined.…”

Section: Resultsmentioning

confidence: 99%

Spatiotemporally Monitoring Cell Viability through Programmable Mitochondrial Membrane Potential Transformation by Using Fluorescent Carbon Dots

et al. 2020

View full text Add to dashboard Cite

Mitochondria are the powerhouse of the cell, which maintain the metabolism of living cells. Mitochondrial membrane potential (MMP) is the key parameter of mitochondria and represents cellular activity. The programmable MMP transformations, i.e., normal, decrease, and vanishing, are indicative mitochondria healthy/damaged/dead states. Therefore, methods for monitoring MMP are of great importance. In this work, carbon dots responsive to mitochondrial membrane potential (MMP‐CDs) are prepared by a simple microwave method. The intracellular distribution of the MMP‐CDs is dependent on the mitochondrial membrane potential. Three different spatiotemporally organelle distributions (mitochondria/lysosome/nucleus) of the MMP‐CDs indicate three distinct MMP levels (normal/decrease/vanishing), which represent three different cell states (healthy/damaged/dead). The normal cells with high mitochondrial membrane potential causes the MMP‐CDs to accumulate in the mitochondria due to the Nernstian effect. The MMP‐CDs are released from the mitochondria and transported to the lysosome when cells are in apoptosis with the decreased mitochondrial membrane potential. In dead cells with a vanished mitochondrial membrane potential, the MMP‐CDs are collocated with the nucleus due to the affinity with DNA. Thus, various cellular states can be visualized through the different localizations of the MMP‐CDs. These MMP‐CDs are successfully employed in the detection of autophagy and H2O2‐induced mitochondria and mitochondrial membrane potential changes.

show abstract

Section: Resultsmentioning

confidence: 99%

Spatiotemporally Monitoring Cell Viability through Programmable Mitochondrial Membrane Potential Transformation by Using Fluorescent Carbon Dots

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Mitochondrial matrix diffusion has been studied with steady-state anisotropy [21], fluorescence correlation spectroscopy (FCS) [4] and fluorescence recovery after photobleaching (FRAP) [22], and has been linked to complex I deficiency and Leigh’s syndrome. Furthermore, a few matrix-targeting lifetime FMRs have been reported [16],[23],[24]. Changes in mitochondrial membrane fluidity have been studied with steady-state anisotropy and linked to a range of neurodegenerative diseases and ageing [2],[3].…”

Section: Introductionmentioning

confidence: 99%

Targeted fluorescence lifetime probes reveal responsive organelle viscosity and membrane fluidity

et al. 2019

View full text Add to dashboard Cite

The only way to visually observe cellular viscosity, which can greatly influence biological reactions and has been linked to several human diseases, is through viscosity imaging. Imaging cellular viscosity has allowed the mapping of viscosity in cells, and the next frontier is targeted viscosity imaging of organelles and their microenvironments. Here we present a fluorescent molecular rotor/FLIM framework to image both organellar viscosity and membrane fluidity, using a combination of chemical targeting and organelle extraction. For demonstration, we image matrix viscosity and membrane fluidity of mitochondria, which have been linked to human diseases, including Alzheimer’s Disease and Leigh’s syndrome. We find that both are highly dynamic and responsive to small environmental and physiological changes, even under non-pathological conditions. This shows that neither viscosity nor fluidity can be assumed to be fixed and underlines the need for single-cell, and now even single-organelle, imaging.

show abstract

“…In this study, we demonstrated the usefulness of complementary techniques for measuring viscosity of discrete subcellular locations in living cells. Whilst numerous previous attempts to target molecular rotors in living cells have relied on exploiting the unique properties of given organelles 7,8 , our work represents one of the first attempts at producing a generic mechanism to target BODIPY to any selected organelle or structure. We measured the relative viscosities of a host of organelles, as well as showcasing the BG-BODIPY's prospects for measuring absolute viscosity in both membrane-bound and membraneless organelles.…”

Section: Discussionmentioning

confidence: 99%

“…Levitt et al (2009) 6 used two meso-substituted BODIPY based fluors conjugated to hydrophobic groups designed to drive indiscriminate incorporation into the hydrophobic interior of non-aqueous cellular membranes, and discovered that the intracellular environment contains a wide range of viscosities. Several groups have targeted boron-dipyrrin (BODIPY) modified by the addition of triphenylphosphonium to target the dye to the mitochondria with varying degrees of success 7,8 . More recently, Chambers et al (2018) 9 , have developed a chloroalkane -BODIPY capable of targeting to specific organelles via a transiently expressed HALO-tag protein targeted to mitochondria or the ER lumen.…”

Section: Introductionmentioning

confidence: 99%

Mapping viscosity in discrete subcellular locations with a BODIPY based fluorescent probe

Pytowski

Foley

Hernández

et al. 2019

Preprint

View full text Add to dashboard Cite

Numerous cellular processes, including enzyme behaviour, signalling, and protein folding and transport are highly influenced by the local microviscosity environment within living cells. Molecular rotors are fluorescent molecules that respond to the viscosity of their environment through changes in both the intensity and lifetime of their fluorescence. We have synthesised a novel boron-dipyrrin (BODIPY) molecular rotor that is also a substrate for the SNAP-tag targeting system (named BG-BODIPY), allowing us to target the rotor to discrete locations within the living cell. We demonstrate that BG-BODIPY reports viscosity, and that this can be measured either through fluorescence lifetime or intensity ratiometric measurements. The relative microviscosities within the ER, Golgi, mitochondrial matrix, peroxisomes, lysosomes, cytoplasm, and nucleoplasm were significantly different. Additionally, this approach permitted fluorescence lifetime imaging microscopy (FLIM) to determine the absolute viscosity within both mitochondria and stress granules, showcasing BG-BODIPY's usefulness in studying both membrane bound and membraneless organelles. These results highlight targeted BG-BODIPY's broad usefulness for making measurements of cellular viscosity both with FLIM and conventional confocal microscopy, the latter option greatly extending the accessibility of the technique.

show abstract

Targetable and fixable rotor for quantifying mitochondrial viscosity of living cells by fluorescence lifetime imaging

Abstract: A fixable probe, named Vis-A, to quantify mitochondrial viscosity of living cells by fluorescence lifetime imaging.

Cited by 92 publications

References 59 publications

Spatiotemporally Monitoring Cell Viability through Programmable Mitochondrial Membrane Potential Transformation by Using Fluorescent Carbon Dots

Spatiotemporally Monitoring Cell Viability through Programmable Mitochondrial Membrane Potential Transformation by Using Fluorescent Carbon Dots

Targeted fluorescence lifetime probes reveal responsive organelle viscosity and membrane fluidity

Mapping viscosity in discrete subcellular locations with a BODIPY based fluorescent probe

Contact Info

Product

Resources

About