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Tuesday, July 28, 2020 | History

3 edition of Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy found in the catalog.

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy

Loling Song

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy

by Loling Song

  • 371 Want to read
  • 1 Currently reading

Published by Delft University Press in Delft .
Written in English

    Subjects:
  • Fluorescence microscopy,
  • Photochemistry,
  • Fluorescein

  • Edition Notes

    Statementdoor Loling Song.
    The Physical Object
    Paginationix, 110 p. :
    Number of Pages110
    ID Numbers
    Open LibraryOL17037502M
    ISBN 109040712220
    OCLC/WorldCa35280326

    Photobleaching is a major factor limiting the observation time in fluorescence microscopy. We achieve photobleaching reduction in structured illumination microscopy (SIM) by locally adjusting the illumination intensities according to the sample. Adaptive SIM is enabled by a digital micro-mirror device (DMD), which provides a projection of the grayscale illumination patterns. Total internal reflection fluorescence microscopy (TIRFM) is an important method in surface science and for the analysis of surface-bound macromolecules. Here, we developed and explored the use of a novel fluorescein isothiocyanate isomer I (FITC)-adsorbed monolayer for alignment and validation of TIRFM measurements and configurations. Aqueous solutions of FITC exist as several different.

    Enter the password to open this PDF file: Cancel OK. File name: . Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy By L. (author) Song, E.J. (author) Hennink, I.T. (author) Young and H.J. (author) Tanke Publisher: Biophysical Society.

    adshelp[at] The ADS is operated by the Smithsonian Astrophysical Observatory under NASA Cooperative Agreement NNX16AC86A.   Books. Publishing Support. Login. Young T and Tanke H J Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy Biophysical J. 68 Crossref [3] Soumpasis D M Theoretical analysis of fluorescence photobleaching recovery experiments Biophysical J. 41


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Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy by Loling Song Download PDF EPUB FB2

The individual photochemical reactions of fluorescein were incorporated into a theoretical analysis and mathematical simulation to study the photochemical processes leading to photobleaching of fluorescein in microscopy.

The photobleaching behavior of free and bound fluorescein has also been investigated by experimental by: Biophysical Journal Volume68 June Photobleaching Kinetics of Fluorescein in Quantitative Fluorescence Microscopy Loling Song,*f E. Hennink,*§ 1. TedYoung,* and HansJ. Tanke* *Department of Cytochemistry and Cytometry, Faculty of Medicine, Leiden University, ALLeiden; *Departmentof Pattern Recognition, Faculty of Applied Physics, Delft University of File Size: 1MB.

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy. L Song, E J Hennink, I T Young, and H J Tanke Department of Cytochemistry and Cytometry, Faculty of Medicine, Leiden University, The Netherlands.

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy. Biophys J. Jun; 68 (6)– [PMC free article] [Google Scholar] Associated Data Supplementary Materials Articles from Biophysical Journal are provided here Cited by: Giloh, H. and Sedat, J.

Fluorescence microscopy: Reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science. Freddy Adams, Carlo Barbante, in Comprehensive Analytical Chemistry, Photobleaching and Fluorescence Lifetime Imaging Microscopy.

Photobleaching complicates the observation of fluorescent molecules, since they will eventually be destroyed by the light exposure necessary to stimulate them into fluorescing.

It can, however, be used to reveal the location and movement of. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by. Logging of fluorescence properties and photobleaching kinetics.

To measure the amount of absorbed photons per excitation event (multiphoton-order) in a regime with no recognizable photobleaching, fluorescence intensity I versus mean power P was recorded for several powers at the same position of the sample.

The obtained data was fitted exponentially, giving the resulting. Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy.

Biophys J. Jun; 68 (6)– [PMC free article] Articles from Biophysical Journal are provided here courtesy of The Biophysical Society. Light-sheet microscopy has been widely used in high-speed fluorescence imaging with low phototoxicity, while the trade-off between the field-of-view and optical sectioning capability limits its application in large-scale imaging.

Although Bessel beam light-sheet microscopy greatly enhances the light-sheet length with the self-healing ability, it suffers from the strong side-lobe effect. Similarly for the two-photon excitation case, the log-log plot of the initial fluorescence intensity versus power (open squares in Fig.

2 B) gave the expected slope for two-photon excitation (slope = ± ).However, the log-log plot of the fluorescein dextran photobleaching rate under two-photon excitation ( nm) gave a slope of ± (filled squares in Fig. 2 B), indicating that. Photobleaching and related photochemical processes are recognized experimental barriers to quantification of fluorescence by microscopy.

We have measured the kinetics of photobleaching of fluorophores in living and fixed cells and in microemulsions, and have demonstrated the spatial variability of these processes within individual cells.

where I [W] is the fluorescence intensity emitted from the fluorescent material per unit time, I 0 [W] is the excitation light flux of the fluorescent material, Φ is the fluorescence quantum yield, C [mol/L] is the concentration of the fluorescent dye, ε is the absorbance index [L/molcm], x [cm] is the diameter of the hydrogel fluorescence microsensor, t [s] is the total exposure time, and.

Photobleaching Kinetics of Fluorescein in Quantitative Fluorescence Microscopy. of fluorescein in microscopy. The photobleaching behavior of free and bound fluorescein has also been. Song L, Hennink E, Young I T and Tanke H J Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy Biophys.

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy. Biophys J. Jun; 68 (6)– [PMC free article] Song L, Varma CA, Verhoeven JW, Tanke HJ.

Influence of the triplet excited state on the photobleaching kinetics of fluorescein in microscopy. image quality of scanning confocal microscopy [37, 38], two-photon microscopy [21, 39] and super-resolutionimaging[40,41].

In contrast, fluorescence recovery after photo-bleaching (FRAP) [42, 43] and fluorescence loss in photobleaching (FLIP) [44] are technically based on the application of low-photostable fluorescent dyes [45, 46]. Song L, Hennink E J, Young I T and Tanke H J Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy Biophys.

68 – Crossref. Limits of Fluorescence as Measure of Probe Concentration As pointed out in the previous section, one of the major goals of quantitative fluorescence microscopy is to determine the re lative amount of tagged molecules in individual intracellular organelles for varying conditions and eventually over time.

Fluorescence microscopy of living cells enables visualization of the dynamics and interactions of intracellular molecules. However, fluorescence live-cell imaging is limited by photobleaching and. Demands of higher spatial and temporal resolutions in linear and nonlinear imaging keep pushing the limits of optical microscopy.

We showed recently that a multiphoton microscope with kHz repetition rate and wide-field illumination has a 2–3 orders of magnitude improved throughput compared to a high repetition rate confocal scanning microscope.

An experimental set up, developed in house, was employed for recording fluorescence spectra from the urine samples. Fig. 1 shows the experimental set up for fluorescence spectral measurement. It uses a nm diode laser (of ∼ mW optical power) for sample illumination and a spectrometer for detecting the fluorescence backscattered from the sample kept in a quartz cuvette.

The studies of fluorescein photobleaching kinetics in microscopy have shown that in general this is not a single-exponential process, and such behavior is caused by the oxygen-independent, proximity-induced triplet-triplet or triplet-ground state dye reactions [77, ]. The single-exponential kinetics when the bleaching follows the first-order.