Search results for "fluorescence lifetime"
showing 10 items of 17 documents
Polysorbate 80 controls Morphology, structure and stability of human insulin Amyloid-Like spherulites
2022
AbstractAmyloid protein aggregates are not only associated with neurodegenerative diseases and may also occur as unwanted by-products in protein-based therapeutics. Surfactants are often employed to stabilize protein formulations and reduce the risk of aggregation. However, surfactants alter protein-protein interactions and may thus modulate the physicochemical characteristics of any aggregates formed. Human insulin aggregation was induced at low pH in the presence of varying concentrations of the surfactant polysorbate 80. Various spectroscopic and imaging methods were used to study the aggregation kinetics, as well as structure and morphology of the formed aggregates. Molecular dynamics s…
Corrigendum to “Polysorbate 80 controls Morphology, structure and stability of human insulin Amyloid-Like spherulites” [J. Colloid Interface Sci. 606…
2023
Direct investigation of viscosity of an atypical inner membrane of Bacillus spores: A molecular rotor/FLIM study
2013
Abstract We utilize the fluorescent molecular rotor Bodipy-C12 to investigate the viscoelastic properties of hydrophobic layers of bacterial spores Bacillus subtilis. The molecular rotor shows a marked increase in fluorescence lifetime, from 0.3 to 4 ns, upon viscosity increase from 1 to 1500 cP and can be incorporated into the hydrophobic layers within the spores from dormant state through to germination. We use fluorescence lifetime imaging microscopy to visualize the viscosity inside different compartments of the bacterial spore in order to investigate the inner membrane and relate its compaction to the extreme resistance observed during exposure of spores to toxic chemicals. We demonstr…
On the Effect of Downscaling in Inkjet Printed Life-Inspired Compartments
2019
The fabrication of size-scalable liquid compartments is a topic of fundamental importance in synthetic biology, aiming to mimic the structures and the functions of cellular compartments. Here, inkjet printing is demonstrated as a customizable approach to fabricate aqueous compartments at different size regimes (from nanoliter to femtoliter scale) revealing the crucial role of size in governing the emerging of new properties. At first, inkjet printing is shown to produce homogenous aqueous compartments stabilized by oil-confinement with mild surfactants down to the hundreds of picoliter scale [1]. Raster Image Correlation Spectroscopy allows to monitor few intermolecular events by the involv…
Blue autofluorescence in protein aggregates “lighted on” by UV induced oxidation
2019
Oxidation of amino acid side chains in protein structure can be induced by UV irradiation leading to critical changes in molecular structure possibly modifying protein stability and bioactivity. Here we show, by using a combination of multiple spectroscopic techniques and Fluorescence Lifetime Imaging, that UV-light exposure induces irreversible oxidation processes in Ubiquitin structure. In particular, the growth of a new autofluorescence peak in the blue region is detected, that we attribute to tyrosine oxidation products. Blue autofluorescence intensity is found to progressively increase also during aggregation processes leading to the formation of aggregates of non-amyloid nature. Signi…
Development of multidimensional spectroscopic and microscopic techniques to facilitate the monitoring of native fluorescence of biomolecules.
2022
There is a need for the development of rapid and reliable characterization tools for biological media. The objective of this thesis is to develop a method based on the acquisition of excitation-emission matrices of fluorescence (EEMF) coupled with the use of fluorescence lifetime measurement in spectroscopy and microscopy (FLIM). These techniques have great potential due to their speed, low sample volume required for analysis, non-destructive sample analysis, and low cost. This project focused on two biological media of great interest to the food industry: wine and bacterial spores. On one hand, we have a beverage representing a large world market, and on the other hand, a food contaminant …
Probing ensemble polymorphism and single aggregate structural heterogeneity in insulin amyloid self-assembly.
2020
Ensembles of protein aggregates are characterized by a nano- and micro-scale heterogeneity of the species. This diversity translates into a variety of effects that protein aggregates may have in biological systems, both in connection to neurodegenerative diseases and immunogenic risk of protein drug products. Moreover, this naturally occurring variety offers unique opportunities in the field of protein-based biomaterials. In the above-mentioned fields, the isolation and structural analysis of the different amyloid types within the same ensemble remain a priority, still representing a significant experimental challenge. Here we address such complexity in the case of insulin for its relevance…
Phasor-FLIM analysis of Thioflavin T self-quenching in Concanavalin amyloid fibrils
2020
The formation of amyloid structures has traditionally been related to human neurodegenerative pathologies and, in recent years, the interest in these highly stable nanostructures was extended to biomaterial sciences. A common method to monitor amyloid growth is the analysis of Thioflavin T fluorescence. The use of this highly selective dye, diffused worldwide, allows mechanistic studies of supramolecular assemblies also giving back important insight on the structure of these aggregates. Here we present experimental evidence of self-quenching effect of Thioflavin T in presence of amyloid fibrils. A significant reduction of fluorescence lifetime of this dye which is not related to the propert…
Fluorescence and spin properties of defects in single digit nanodiamonds
2009
International audience; This article reports stable photoluminescence and high-contrast optically detected electron spin resonance (ODESR) from single nitrogen-vacancy (NV) defect centers created within ultrasmall, disperse nanodiamonds of radius less than 4 nm. Unexpectedly, the efficiency for the production of NV fluorescent defects by electron irradiation is found to be independent of the size of the nanocrystals. Fluorescence lifetime imaging shows lifetimes with a mean value of around 17 ns, only slightly longer than the bulk value of the defects. After proper surface cleaning, the dephasing times of the electron spin resonance in the nanocrystals approach values of some microseconds, …
Single molecules probe local density of modes (LDOS) around photonic nanostructures
2008
International audience; According to Fermi's golden rule, the fluorescence decay rate is directly proportional to the projected local density of photonic modes (LDOS) at the molecule location. The relevant LDOS depends on the molecule orientation. In this paper, the direct measurement of the fluorescence lifetime near gold dot photonic structures is investigated and compared to calculated LDOS. Detailed analysis of the decay channels is presented on the basis of numerical simulations.