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Improved FRAP Measurements on Biofilms.

ABSTRACT: We expand the standard fluorescence recovery after photobleaching (FRAP) model introduced by Axelrod et al. in 1976. Our goal is to capture some of the following common artifacts observed in the fluorescence measurements obtained with a confocal laser scanning microscope in biofilms: 1) linear drift, 2) exponential decrease (due to bleaching during the measurements), 3) stochastic Gaussian noise, and 4) uncertainty in the exact time point of the onset of fluorescence recovery. To fit the resulting stochastic model to data from FRAP measurements and to estimate all unknown model parameters, we apply a suitably adapted Metropolis-Hastings algorithm. In this way, a more accurate estimation of the diffusion coefficient of the fluorophore is achieved. The method was tested on data obtained from FRAP measurements on a cultivated biofilm.

SUBMITTER: Hauth J

PROVIDER: S-EPMC7231900 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Improved FRAP Measurements on Biofilms.

Hauth Jan J Chodorski Jonas J Wirsen Andreas A Ulber Roland R

Biophysical journal 20200404 10

We expand the standard fluorescence recovery after photobleaching (FRAP) model introduced by Axelrod et al. in 1976. Our goal is to capture some of the following common artifacts observed in the fluorescence measurements obtained with a confocal laser scanning microscope in biofilms: 1) linear drift, 2) exponential decrease (due to bleaching during the measurements), 3) stochastic Gaussian noise, and 4) uncertainty in the exact time point of the onset of fluorescence recovery. To fit the resulti ...[more]

PMID: 32304636

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Project description:Fricke solution has a wide range of applications as radiation detector and dosimetry. It is particularly appreciated in terms of relevant comparative advantages, like tissue-equivalence when prepared in aqueous media like gel matrix, continuous mapping capability, independence of dose rate and incident direction, as well as linear dose response. This work presents the development and characterization of an improved Fricke gel system, based on modified chemical compositions, making possible its application in clinical radiology due to its improved sensitivity. Properties of standard Fricke gel dosimeter for high-dose levels are used as a starting point, and suitable chemical modifications are introduced and carefully investigated in order to attain high resolution for low-dose ranges, like those corresponding to radiology interventions. The developed Fricke gel radiation dosimeter system achieves the expected typical dose-dependency, showing linear response in the dose range from 20 up to 4000 mGy. Systematic investigations including several chemical compositions are carried out in order to obtain an adequate dosimeter response for low-dose levels. A suitable composition from among those studied is selected as a good candidate for low-dose-level radiation dosimetry consisting of a modified Fricke solution fixed to a gel matrix containing benzoic acid along with sulfuric acid, ferrous sulfate, Xylenol orange, and tridistilled water. Dosimeter samples are prepared in standard vials for in-phantom irradiation and further characterization by spectrophotometry measuring visible light transmission and absorbance before and after irradiation. Samples are irradiated using typical X-ray tubes for radiology and calibrated Farmer-type ionization chamber is used as reference to measure dose rates inside phantoms at vial locations. Once sensitive material composition is optimized, dose-response curves show significant improvement regarding overall sensitivity for low dose levels. The aim of this work consists of implementing the optimized gel dosimeter to perform direct measurements of absorbed dose in samples irradiated during microcomputed tomography scanning in order to preliminary assess dose levels for further scanning of small animals for further applications in veterinary and paleontology. As a first attempt, dose distributions were measured in water-equivalent phantoms having dimensions comparable to small animals, 100 to 1000 cm3, approximately. According to the obtained results, it is found that the proposed method shows satisfactory reliability and adequate performance for a promising gel dosimetry system.

Dataset Information

Improved FRAP Measurements on Biofilms.

Publications

Improved FRAP Measurements on Biofilms.

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