Peer Reviewed: Applications of Fluorescence Recovery after Photobleaching

Kovaleski, John M.; Wirth, Mary J.

doi:10.1021/ac971792r

Cited by 19 publications

(8 citation statements)

References 31 publications

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“…This microscopic, non-destructive and slightly invasive technique, in which the probe concentration remains micromolar, originates from mobility studies in biological membranes [5]. It was then extended to other fields, mostly for liquid or highly hydrated systems, in which diffusion follows the Stokes-Einstein law [44]. It covers a wide range of apparent diffusion coefficients, from 10 −20 to 10 −9 m 2 ·s −1 [43].…”

Section: Fluorescence Recovery After Photobleachingmentioning

confidence: 99%

Determination of the diffusion coefficients of small solutes in cheese: A review

Floury

Jeanson

Aly

et al. 2010

Dairy Sci. Technol.

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-In cheese technology, the mass transfer of small solutes, such as salt, moisture and metabolites during brining and ripening, is very important for the final quality of the cheese. This paper has the following objectives: (i) to review the data concerning the diffusion coefficients of solutes in different cheese types; (ii) to review the experimental methods available to model the mass transfer properties of small solutes in complex matrices such as cheese; and (iii) to consider some potential alternative approaches. Numerous studies have reported the transfer of salt in cheese during brining and ripening. Regardless of the type of cheese and its composition, the effective diffusion coefficients of salt have been reported to be between 1 and 5.3 × 10 −10 m 2 ·s −1 at 10-15°C. However, few papers have dealt with the mass transfer properties of other small solutes in cheese. Most of the reported effective diffusion coefficient values have been obtained by macroscopic and destructive concentration profile methods. More recently, some other promising techniques, such as nuclear magnetic resonance, magnetic resonance imaging or fluorescence recovery after photobleaching, are currently being developed to measure the mass transfer properties of solutes in heterogeneous media at microscopic scales. However, these methods are still difficult to apply to complex matrices such as cheese. Further research needs to focus on: (i) the development of nondestructive techniques to determine the mass transfer properties of small solutes at a microscopic level in complex matrices such as cheese; and (ii) the determination of the mass transfer properties of metabolites that are involved in enzymatic reactions during cheese ripening. cheese / mass transfer / diffusion / modelling / solute Article published by EDP SciencesRésumé -Détermination des coefficients de diffusion de petits solutés dans le fromage : une synthèse. En technologie fromagère, le transfert de petits solutés, tels que le sel, l'eau et les métabolites au cours du saumurage et de l'affinage, joue un rôle majeur sur la qualité finale du fromage. Cette revue bibliographique a pour objectifs principaux : (i) de faire le bilan des valeurs publiées des coefficients de diffusion de différents solutés dans les fromages ; (ii) de passer en revue les méthodes expérimentales disponibles pour déterminer les propriétés de transfert des petits solutés dans des milieux complexes comme le fromage ; (iii) de considérer les méthodes alternatives potentiellement applicables aux fromages. Dans la littérature, de nombreuses études ont été publiées au sujet du transfert de sel dans les fromages au cours du saumurage et de l'affinage. En fonction du type de fromage et de sa composition, les coefficients de diffusion effectifs du sel sont compris entre 1 et 5,3 × 10 −10 m 2 ·s −1 à des températures comprises entre 10 et 15°C. Très peu d'études concernant les propriétés de transfert d'autres petits solutés dans les fromages ont été publiées. La plupart des coefficients de diffusi...

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Section: Fluorescence Recovery After Photobleachingmentioning

confidence: 99%

Determination of the diffusion coefficients of small solutes in cheese: A review

Floury

Jeanson

Aly

et al. 2010

Dairy Sci. Technol.

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“…FPR methods have been reviewed recently. [36][37][38][39][40][41][42] In an ideal FPR measurement, some fraction of covalently attached dyes are destroyed without damaging the macromolecule or heating the solution. The recovery provides information about the diffusion.…”

Section: Introductionmentioning

confidence: 99%

Matrix Fluorescence Photobleaching Recovery for Polymer Molecular Weight Distributions and Other Applications

et al. 2006

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A new method for determining molecular weight distributions is explored. Matrix fluorescence photobleaching recovery (MFPR) requires dye attachment to the polymer analyte. No physical separation of macromolecules is accomplished, as in the predominant gel permeation chromatography technique. Rather, the distribution of diffusion coefficients is determined by inverse Laplace transformation of fluorescence photobleaching recovery decay profiles, and the diffusion data are then mapped onto molecular weight using a measured calibration. Resolution is enhanced by forcing the labeled polymer analyte to diffuse through a solution that contains unlabeled matrix, which can even be the same type of polymer. Relative to free diffusion, this increases the dependence of diffusion on molecular weight, making it easier to differentiate components having similar mass. Simulated, noise-corrupted data for molecules diffusing by reptation suggest that excellent resolution may be achieved. Empirical tests were conducted using fluorescent dextran and pullulan diffusers in unlabeled dextran matrix solutions. Although the resolution fell short of the simulated results, it exceeded that available from gel permeation chromatography as normally practiced. An investigation conducted to understand why the full resolution was not achieved revealed that matrix solutions of high molecular weight dextran lie in the unentangled semidilute regime, as evidenced by the scaling behavior of probe diffusion with probe molecular weight, the absence of a rheological plateau modulus, and the shape of the small-angle X-ray scattering profiles. Branching of high-molecular-weight dextran seems to preclude entry into the entangled regime, where a stronger diffusion vs mass relationship is expected. The good performance that was nevertheless achieved suggests that further exploration may prove fruitful.

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“…There has been a considerable amount of interest in the lateral diffusion of adsorbates at chemical interfaces for a variety of important systems, including biological membranes, artificial membranes, , chromatographic interfaces, − and liquid−liquid interfaces. − Lateral diffusion is measured very accurately by either fluorescence recovery after photobleaching − ,− or fluorescence correlation spectroscopy, − where the latter avoids perturbing the sample. The interpretation of the results is most straightforward when there is a single diffusing species; however, the data can, in principle, be analyzed for multiple species. , …”

Section: Introductionmentioning

confidence: 99%

Single-Molecule Study of an Adsorbed Oligonucleotide Undergoing Both Lateral Diffusion and Strong Adsorption

Wirth

Swinton

2001

J. Phys. Chem. B

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Single-molecule fluorescence spectroscopy was used to probe the lateral transport of an adsorbed oligonucleotide, the SP6 promoter primer, which is a 24-mer labeled at the 5′ end with tetramethylrhodamine. The oligonucleotide was adsorbed to the interface of an aqueous solution of 0.01 M KCl and silica chemically modified by chlorodimethyloctadecylsilane. Confocal fluorescence microscopy achieved single-molecule resolution, with a molecule in the beam 7% of the time. Autocorrelation of the data fit well to a model having two species, one diffusing and the other one undergoing transient strong adsorption. A small number of bursts, 0.3%, had unusually long durations, accounting for the slow component in the autocorrelation. When these long bursts were excised from the data, the autocorrelation fit well to simple diffusion, with D ) 4 × 10 -6 cm 2 /s. Autocorrelation of the long bursts alone gave a rate constant for desorption of 3 s -1 . The strongly adsorbed molecules were found to comprise 10% of the total population of adsorbates. It is concluded that the lateral transport of the SP6 promoter primer is described by fast lateral diffusion interrupted by rare, reversible, strong adsorption to defect sites. During strong adsorption, the tetramethylrhodamine label undergoes hindered motion, suggesting it is not the adsorbing moiety.

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Peer Reviewed: Applications of Fluorescence Recovery after Photobleaching

Cited by 19 publications

References 31 publications

Determination of the diffusion coefficients of small solutes in cheese: A review

Determination of the diffusion coefficients of small solutes in cheese: A review

Matrix Fluorescence Photobleaching Recovery for Polymer Molecular Weight Distributions and Other Applications

Single-Molecule Study of an Adsorbed Oligonucleotide Undergoing Both Lateral Diffusion and Strong Adsorption

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