Multiscale Live Imaging Using Förster Resonance Energy Transfer (FRET) for Evaluating the Biological Behavior of Nanoparticles as Drug Carriers

Ishizawa, Kiyomi; Togami, Kohei; Tada, Hitoshi; Chono, Sumio

doi:10.1016/j.xphs.2020.08.028

Cited by 7 publications

(3 citation statements)

References 46 publications

(63 reference statements)

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“…Genetically encoded fluorescent proteins (FPs)-based Förster resonance energy transfer (FRET) technology has been widely used for mapping temporal-spatial dynamics of intracellular biochemical events in living cells [1][2][3][4][5][6][7][8][9][10][11] "3-cube FRET" microscopy, referred to 3-cube FRET method is the most extensively applied approach for live-cell FRET quantification. [12][13][14][15] Three images are needed for E-FRET image: I DD (DA), image from donor channel with donor excitation; I AA (DA), image from acceptor channel with acceptor excitation; and I DA (DA), image from acceptor channel with donor excitation.…”

Section: Introductionmentioning

confidence: 99%

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

et al. 2021

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Three-cube Förster resonance energy transfer (FRET) method is the most extensively applied approach for live-cell FRET quantification. Reliable measurements of calibration factors are crucial for quantitative FRET measurement. We here proposed a modified TA-G method (termed as mTA-G) to simultaneously obtain the FRETsensitized quenching transition factor (G) and extinction coefficients ratio (γ) between donor and acceptor. mTA-G method includes four steps: ( 1) predetermining the ratio ranges of the sensitized emission of acceptor (F C ) to the donor excitation and donor channel image (I DD [(DA])) for all FRET plasmids; (2) culturing the cells which express every FRET plasmid in one dish respectively; (3) distinguishing and marking the cells expressing different FRET plasmids by detecting their F C /I DD (DA) values; (4) linearly fitting F C /I AA (DA) (acceptor excitation and acceptor channel image) to I DD (DA)/I AA (DA)for different kinds of cells. We implemented mTA-G method by imaging tandem constructs cells with different FRET efficiency cultured in one dish on different days, and obtained consistent G and γ values. mTA-G method not only circumvents switchover of different culture dishes but also keep the constant imaging conditions, exhibiting excellent robustness, and thus will expands the biological applications of quantitative FRET analysis in living cells.

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Section: Introductionmentioning

confidence: 99%

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

et al. 2021

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“…An understanding of FRET in the sheet regime and how it is affected by various nonidealities is not just an academic exercise, but can inform real-world applications. Sheet regime effects become critical for quantitatively interpreting FRET microscopy of cellular membranes where one typically monitors dye-labeled D moieties interacting with membranes labeled with numerous lipophilic A dyes (or displaying some endogenous receptor/protein that can similarly act as the A sheet) in order to gather information about ligand–receptor interaction dynamics, vesicle formation, membrane biomolecular sensing events, etc . ,− The density of fluorophores in such confines can make interpretation of ET and subsequent distance estimates complex. Moreover, it is in just such situations where changing the 1/ r α exponent value from 6 to 4 can significantly alter estimated separation distances and assumptions of viable FRET sensing ranges.…”

Section: Discussionmentioning

confidence: 99%

“…Our goal in this paper is to better understand some of the key variables that enable Förster transfer in the sheet regime in real systems. Our focus is on systems of discrete donors and acceptors and not on physically continuous systems such as graphene, metallic absorbers, or surfaces, other 2D materials, or larger (metallic) plasmonic nanoparticle surfaces whose optoelectronic properties are still not fully characterized and where more complex processes may potentially be involved. − Beyond designing improved FRET networks, this work is also relevant to FRET-based applications such as fluorescent imaging and characterization of cellular membranes and probing of ligand–receptor interactions. ,− …”

Section: Introductionmentioning

confidence: 99%

Understanding Förster Resonance Energy Transfer in the Sheet Regime with DNA Brick-Based Dye Networks

Mathur

Samanta

Ancona³

et al. 2021

ACS Nano

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Controlling excitonic energy transfer at the molecular level is a key requirement for transitioning nanophotonics research to viable devices with the main inspiration coming from biological light-harvesting antennas that collect and direct light energy with near-unity efficiency using Förster resonance energy transfer (FRET). Among putative FRET processes, point-to-plane FRET between donors and acceptors arrayed in two-dimensional sheets is predicted to be particularly efficient with a theoretical 1/r 4 energy transfer distance (r) dependency versus the 1/r 6 dependency seen for a single donor–acceptor interaction. However, quantitative validation has been confounded by a lack of robust experimental approaches that can rigidly place dyes in the required nanoscale arrangements. To create such assemblies, we utilize a DNA brick scaffold, referred to as a DNA block, which incorporates up to five two-dimensional planes with each displaying from 1 to 12 copies of five different donor, acceptor, or intermediary relay dyes. Nanostructure characterization along with steady-state and time-resolved spectroscopic data were combined with molecular dynamics modeling and detailed numerical simulations to compare the energy transfer efficiencies observed in the experimental DNA block assemblies to theoretical expectations. Overall, we demonstrate clear signatures of sheet regime FRET, and from this we provide a better understanding of what is needed to realize the benefits of such energy transfer in artificial dye networks along with FRET-based sensing and imaging.

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Advanced molecular imaging for the characterisation of complex medicines

Yilmaz

Sharp

Main

et al. 2022

Drug Discovery Today

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Multiscale Live Imaging Using Förster Resonance Energy Transfer (FRET) for Evaluating the Biological Behavior of Nanoparticles as Drug Carriers

Cited by 7 publications

References 46 publications

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

Understanding Förster Resonance Energy Transfer in the Sheet Regime with DNA Brick-Based Dye Networks

Advanced molecular imaging for the characterisation of complex medicines

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