Technique for the determination of fluorescence quantum efficiencies: a method avoiding direct measurement of absorbance

Britten, A. Z.; Archer-Hall, John; Lockwood, G. G.

doi:10.1039/an9780300928

Cited by 16 publications

(9 citation statements)

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“…Several reports have appeared on methods for determining the quantum yield of fluorescence and phosphorescence of organic compounds in solution (1)(2)(3)(4)(5). Substantially less work has been reported for determining the luminescence quantum yield for solid materials.…”

mentioning

confidence: 99%

Determination of room-temperature fluorescence and phosphorescence quantum yields for compounds adsorbed on solid surfaces

Ramasamy¹,

Senthilnathan²,

Hurtubise³

1986

Anal. Chem.

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mentioning

confidence: 99%

Determination of room-temperature fluorescence and phosphorescence quantum yields for compounds adsorbed on solid surfaces

Ramasamy¹,

Senthilnathan²,

Hurtubise³

1986

Anal. Chem.

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“…The brightness correlates with the extinction coefficient and the quantum yield. Whereas, the extinction coefficient describes the ability of a dye to absorb photons of a certain wavelength, the quantum yield determines the ratio of absorbed to emitted photons [13]. If a dye is not sufficiently bright it influences the SNR negatively.…”

Section: Fluorescent Dyes Are the Most Important Components In Fcsmentioning

confidence: 99%

Ligands for Fluorescence Correlation Spectroscopy on G Protein-Coupled Receptors

Jakobs

Sorkalla²,

Häberlein³

2012

CMC

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G protein-coupled receptors (GPCRs) comprise a large protein family of transmembrane receptors involved in many physiological processes. They are engaged in various transduction processes of extracellular signals into intracellular responses. Due to their involvement in numerous diseases they represent an important pharmacological target. Fluorescence correlation spectroscopy (FCS) poses a very sensitive analytical technique well-suited for the investigation of GPCRs. It is minimally invasive and operates on a single molecular level. It further provides detailed pharmacological information on receptor kinetics and quantities of activated receptors on the cell membrane. In addition, FCS allows distinguishing between different receptor states based on different diffusion time constants. In order to be applicable for FCS, the molecule of interest has to be fluorescently labeled. This review focuses on the physical requirements for dyes intended for FCS, their influence on the binding characteristics of coupled ligands and strategies to generate dye labeled ligands, exemplified on GPCR ligands.

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“…The corrected radiant power Po is then determined by substituting cc and the experimentally observed Pb into eq 8, which follows from eq 5 and 6 Po = 2.303Pbebc/(le-2.303ebe) (8) EXPERIMENTAL SECTION Apparatus. To investigate this model, we designed a cell and collimator to allow the measurement of the ratios of CL from a homogeneous solution at two pathlengths as shown in Figure 2.…”

Section: ~Be)/(2303~c) ( 5 )mentioning

confidence: 99%

Absorption-corrected chemiluminescence measurements with a dual-pathlength spectrometer

Ratzlaff¹,

Crouch²

1983

Anal. Chem.

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typically 5-15 min. Light scattering and absorption by the support can affect and may distort the apparent intensities of the peaks in the spectrum, so care must be taken when attempting to do quantitative work.A mathematical model of secondary inner-fitter effects allows the derivation of a correction factor based on a determination of the ratio of chemiluminescence intensities from two pathlengths. A microprocessor-controlled instrument equipped with a unlque dual-path cell acquires intenstty measurements from a collimated, monochromatic optical system. Lumlnol chemiluminescence was observed from solutions containing the interfering chromophores picrate and ferroin. The effectiveness of the model and the characteristics and limitations of the instrument are discussed. For the case of negiiglbie reemission, absorption-free signals are accurately calculated for matrices wlth absorbances ranging to more than 0.75. An emission spectrum demonstrating ferroin activation Is recovered from an apparently quenched reaction.displacements of the through-cell paths of the excitation and emission beams (5-8). For a well-defined optical geometry, the fluorescence attenuation changes with the pathlength according to Beer's law. Intensity vs. pathlength information is used to determine the absorbance values, and a correction factor is calculated. No similar remedies have been described for correcting CL measurements for inner-filter effects, although an expression describing CL in terms of absorbance and pathlength has been reported by Stieg and Nieman (9). This expression was applied as a criterion for the efficient design of CL flow cells but not for developing a method to correct the inner-filter effects.In this paper we describe a procedure for correcting CL measurements that is based on a determination of the ratio of CL intensities from two different cell pathlengths. A microprocessor-controlled instrument with a unique dual Dathleneth cell is described. The instrument automaticallv Y produces absorption-corrected CL intensities a t a single wavelength or absorption-corrected CL spectra. Results of studies with a luminal cL reaction are presented that demonstrate the effectiveness of the correction procedure and the characteristics of the instrument. THEORYMany studies Of (cL) and bioluminescence reactions have suggested ambiguities or interferences attributable to inner-filter effects (attenuation of emitted radiant power due to absorption by the solution matrix) (1-3). Methods for correcting molecular fluorescence measurements for inner-filter effects have been reviewed (4, 5). These correction procedures used expressions that describe the fluorescence intensity as a function of the absorbance a t the excitation and/or the emission wavelength. The "cellshift" method for correction of inner-filter effects depends upon fluorescence measurements determined with several Consider a thin slice of solution of infinitesimal thickness dx, parallel to the observation window. The slice radiates monochromatic light of unattenuated ...

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Technique for the determination of fluorescence quantum efficiencies: a method avoiding direct measurement of absorbance

Cited by 16 publications

References 0 publications

Determination of room-temperature fluorescence and phosphorescence quantum yields for compounds adsorbed on solid surfaces

Determination of room-temperature fluorescence and phosphorescence quantum yields for compounds adsorbed on solid surfaces

Ligands for Fluorescence Correlation Spectroscopy on G Protein-Coupled Receptors

Absorption-corrected chemiluminescence measurements with a dual-pathlength spectrometer

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