Polarization-resolved single-molecule tracking reveals strange dynamics of fluorescent tracers through a deep rubbery polymer network

Mahato, Jaladhar; Bhattacharya, Sukanya; Sharma, Dipti; Chowdhury, Arindam

doi:10.1039/d0cp05864e

Cited by 3 publications

(2 citation statements)

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“…Polarization-resolved fluorescence imaging of amyloid fibrils was performed in a wide-field epifluorescence microscope as previously described (Figure 2a). 44 Briefly, ThT-bound amyloid fibrils were spin-cast on glass coverslips and excited by a 405 nm laser (Laserglow, model LRD-0405-PFR-00050-05) circularly polarized light by using a quarter-wave plate. The excitation beam was directed to the sample by using an objective (1.49NA, 60× Nikon, TIRF), which illuminated an 8 × 8 μm 2 FOV.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

et al. 2021

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Amyloid fibrils are structurally heterogeneous protein aggregates that are implicated in a wide range of neurodegenerative and other proteopathic diseases. These fibrils exist in a variety of different tertiary and higher-level structures, and this exhibited polymorphism greatly complicates any structural study of amyloid fibrils. In this work, we demonstrate a method of using polarization-resolved microscopy to directly observe the structural heterogeneity of individual amyloid fibrils using amyloid-bound fluorophores. We formulate a mathematical quantity, helical anisotropy, which utilizes the polarized emission of amyloid-bound fluorophores to report on the local structure of individual fibrils. Using this method, we show how model amyloid fibrils generated from short peptides exhibit diverse structural properties both between different fibrils and within a single fibril, in a manner that is replicated for fibrils assembled from longer proteins. Our method represents an accessible and easily adaptable technique by which polymorphism in the structure of amyloid fibrils can be probed. Additionally, the methodology we describe here can be easily extended to the study of other fibrillar and otherwise ordered supramolecular structures.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

et al. 2021

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“…), the polarization-resolved spectroscopy/imaging techniques provide people with significant tools to retrieve the information of matters. [1][2][3][4][5][6][7][8][9][10] Nowadays, most available polarization-resolved spectroscopy/imaging techniques are realized by scanning the polarization state of incident lights and then plotting/mapping the polarization-resolved spectra/images. However, these shot-to-shot techniques are not suitable for samples with poor optical stability, because irreversible damages may happen in the process of data collecting and further cause the results incomparable from one polarization state to another.…”

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confidence: 99%

Single-scan polarization-resolved saturated absorption spectra using hybridly polarized beam in ⁸⁵Rb atomic vapor

Jiang,

Yuan,

Cheng

et al. 2023

Appl. Phys. Express

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We report on a single-scan polarization-resolved saturated absorption spectra (SAS) method utilizing a hybridly polarized beam as the probe. Owning to the spatial-variant polarization of a hybridly polarized probe, the polarization-resolved information can be retrieved from the single signal image. Then, the polarization dependence of SAS from two transitions are investigated. Strong polarization dependence is observed from the “closed two-level transition” as |5S 1/2, F = 3> → |5P 3/2, F′ = 4> of 85Rb. By contrast, no polarization dependence is observed from “open two-level transition” as |5S 1/2, F = 3> → |5P 3/2, F′ = 3>. The method would be meaningful for the samples with poor stability or the ultrafast processes.

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Single-molecule tracking of dye diffusion in synthetic polymers: A tutorial review

Tran-Ba

Foreman

2023

Journal of Applied Physics

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Single-molecule tracking (SMT) methods have been widely employed to offer a high-resolution characterization of synthetic polymers under ambient conditions and, thus, have advanced our understanding of their unique material properties. SMT is based on the systematic monitoring of the diffusive motions of individual fluorescent dye molecules in the as-prepared polymer thin films or thicker monoliths. Quantitative assessment of the recorded SMT video data involves the systematic analysis of the generated diffusion trajectories of a single molecule using well-established and reported methods. The results have offered a wealth of new information on the structural alignment, orientational order, and long-range continuity of the polymer microdomains; the nanoscale material heterogeneities governed by defects, misalignment, and ill-controlled preparation conditions; as well as the various forms of probe–host interactions on the single-molecule level. In the first part of this Tutorial review, we describe the fundamental principles and instrumentation of SMT, before offering interested readers and potential future SMT users a practical guidance on the selection of fluorescent probe molecules, preparation of suitable samples, and optimization of experimental conditions and imaging parameters. Then, we highlight several representative SMT studies in microphase-separated block copolymers, and semicrystalline and amorphous homopolymers to further emphasize the usefulness of SMT methods for polymer characterization without neglecting some of their shortcomings. This Tutorial review is written with the motivation to attract new researchers to the fast-growing field and assist them in starting their own SMT research of synthetic polymers and other technologically useful soft matter systems.

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Polarization-resolved single-molecule tracking reveals strange dynamics of fluorescent tracers through a deep rubbery polymer network

Abstract: Tracking the movement of fluorescent single-molecule (SM) tracers has provided several new insights on the local structure and dynamics in complex environments such as soft materials and biological systems. However,...

Cited by 3 publications

References 55 publications

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

Single-scan polarization-resolved saturated absorption spectra using hybridly polarized beam in ⁸⁵Rb atomic vapor

Single-molecule tracking of dye diffusion in synthetic polymers: A tutorial review

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Polarization-resolved single-molecule tracking reveals strange dynamics of fluorescent tracers through a deep rubbery polymer network

Abstract: Tracking the movement of fluorescent single-molecule (SM) tracers has provided several new insights on the local structure and dynamics in complex environments such as soft materials and biological systems. However,...

Cited by 3 publications

References 55 publications

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

Investigation of Structural Heterogeneity in Individual Amyloid Fibrils Using Polarization-Resolved Microscopy

Single-scan polarization-resolved saturated absorption spectra using hybridly polarized beam in 85Rb atomic vapor

Single-molecule tracking of dye diffusion in synthetic polymers: A tutorial review

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Product

Resources

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Single-scan polarization-resolved saturated absorption spectra using hybridly polarized beam in ⁸⁵Rb atomic vapor