Absorption-corrected fiber optic fluorometer

Ratzlaff, Eugene H.; Harfmann, Robert G.; Crouch, S. R.

doi:10.1021/ac00267a009

Cited by 23 publications

(15 citation statements)

References 10 publications

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“…The properties of spectral inner filters and analytical expressions to correct for the absorption of excitation radiation (i.e., primary inner filter) and absorption of emitted light (i.e., secondary inner filter) have been described since the mid-1970s, 2 and have recently improved. [3][4][5]7 Although these methods provide a discrete solution for primary and secondary inner-filter effects, they require precise knowledge of the instrumentation geometry and probes to account for these effects. By contrast, FAID is deterministic relying on prior information of the emission and absorption characteristics of the sample alone.…”

Section: Discussionmentioning

confidence: 99%

“…In most cases, this type of analysis requires the decomposition of complex emission spectra due to spectral overlap (for examples, see Refs. [1][2][3][4][5]. Previous approaches have used a linear combination of model emission spectra 1,6 to extract individual fluorophore information.…”

Section: Introductionmentioning

confidence: 99%

“…Several analytical approaches have been developed to correct for these inner-filter effects. [2][3][4][5]7 However, these corrections require extensive prior information, working distances and angles, 2,4,5 and prescribed circumstances such as right-angle cuvettes. 2 Therefore, these approaches generally do not permit the dynamic changes in the sample absorption and scattering characteristics that are common in many biochemical assays.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2

et al. 2007

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A method for decomposing complex emission spectra by correcting for known inner-filter effects is described. This approach builds on previous work using a linear combination of model emission spectra and combines the known absorption characteristics of the system to fit the composite emission spectrum. Rhod-2, which has a small Stokes shift and significant self-absorption, was used as the model system. By adding the absorption characteristics of Rhod-2 to the model, the degree of fit was significantly improved, thus minimizing residuals, and accurately predicted the spectral shape changes with increasing concentration, [Rhod-2]. More complex studies were conducted with Rhod-2 in isolated cardiac mitochondria with multiple emission and absorption elements. By including known absorbances to the spectral decomposition, the overall precision increased almost four fold. Moreover, this approach eliminated the significant [Rhod-2] dependence on the apparent K(50) and therefore improved the accuracy of free [Ca(2+)] calculations. These data demonstrate that secondary inner-filter correction can significantly improve spectral decomposition of complex emission spectra, which are used in a variety of biological applications.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2

et al. 2007

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“…In the traditional literature on fluorescence spectroscopy, the correction of any inner filter effect is mostly associated to the right-angle geometry, based on the assumption that the front-face geometry would not be affected by such a problem, as the measurement would only pertain the surface of the sample. Only a few papers exist dealing with the problem of inner filter effect in front-face fluorescence measurements [38][39][40][41][42] : generally, it is proposed to make a modification of the instrumental apparatus (the instrument or the cell) used to perform the measurement and often make use of several repeated measurement for the analysis of one sample. While most of these papers were published already in the 1970s, the vast majority of the following traditional fluorescence literature mostly neglect this issue and simply present the front-face geometry as one of the possible solutions to the occurrence of inner filter effect in traditional right-angle fluorescence measurements.…”

Section: The Correctionmentioning

confidence: 99%

Investigating challenges with scattering and inner filter effects in front‐face fluorescence by PARAFAC

Bevilacqua

Rinnan

Lund

2020

Journal of Chemometrics

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The use of front‐face fluorescence spectroscopy, and three‐way chemometric analysis (like PARAllel FACtor analysis, PARAFAC), has been successfully applied for the analysis of unpretreated samples in the fields of food and environmental analysis. It would be desirable to evaluate the potential of this approach for the analysis of any real sample in any field of research. Even in the simplest of the real samples acquired with front‐face, the presence of scattering and inner filter effects will occur, potentially hampering the subsequent data analysis due to deviations from Beer‐Lambert's law. This paper addresses these concerns in practice, proposing a strategy of spectral preprocessing to mitigate these effects. This is done by measuring several data sets with different levels of scattering and inner filter effects by fluorescence in excitation‐emission mode. The results show that the occurrence of these interferents (sometimes neglected with front‐face mode) affects the fluorescence signal and interferes with any traditional analysis on these data, as much as they hamper the successful use of methods like PARAFAC. The proposed preprocessing strategy is based on one of the most traditional correction for the inner filter effect with right‐angle mode. However, we suggest applying a tunable factor, b, that will account for the degree of deviation from linearity between concentration of a given analyte and its fluorescence signal. It is demonstrated that by choosing a proper b‐value, this correction helps in finding an acceptable solution for the PARAFAC algorithm, in line with Beer‐Lambert's law.

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“…whether or not the absorbing or scattering species is the species of interest. In the case of fluorescence measurements, several groups examined the effect of IFE and developed analytical or empirical expressions to correct for them (Eisinger and Flores, 1985;Puchalski et al, 1991;Ratzlaff et al, 1984). In each case, the analytical expressions were derived beginning with the Beer-Lambert absorbance theory as a starting point.…”

Section: Inner Filter Effectsmentioning

confidence: 99%

On-line monitoring of intracellular ATP concentration in Escherichia coli fermentations

Lasko

Wang

2000

Biotechnol. Bioeng.

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Absorption-corrected fiber optic fluorometer

Cited by 23 publications

References 10 publications

Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2

Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2

Investigating challenges with scattering and inner filter effects in front‐face fluorescence by PARAFAC

On-line monitoring of intracellular ATP concentration in Escherichia coli fermentations

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Absorption-corrected fiber optic fluorometer

Cited by 23 publications

References 10 publications

Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca2+ Using Rhod-2

Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca2+ Using Rhod-2

Investigating challenges with scattering and inner filter effects in front‐face fluorescence by PARAFAC

On-line monitoring of intracellular ATP concentration in Escherichia coli fermentations

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Product

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Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2

Fluorescence Absorbance Inner-Filter Decomposition: The Role of Emission Shape on Estimates of Free Ca²⁺ Using Rhod-2