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Prediction of intracellular storage polymers using quantitative image analysis in enhanced biological phosphorus removal systems.

Analytica chimica acta (2013-03-19)
Daniela P Mesquita, Cristiano Leal, Jorge R Cunha, Adrian Oehmen, A Luís Amaral, Maria A M Reis, Eugénio C Ferreira
ABSTRACT

The present study focuses on predicting the concentration of intracellular storage polymers in enhanced biological phosphorus removal (EBPR) systems. For that purpose, quantitative image analysis techniques were developed for determining the intracellular concentrations of PHA (PHB and PHV) with Nile blue and glycogen with aniline blue staining. Partial least squares (PLS) were used to predict the standard analytical values of these polymers by the proposed methodology. Identification of the aerobic and anaerobic stages proved to be crucial for improving the assessment of PHA, PHB and PHV intracellular concentrations. Current Nile blue based methodology can be seen as a feasible starting point for further enhancement. Glycogen detection based on the developed aniline blue staining methodology combined with the image analysis data proved to be a promising technique, toward the elimination of the need for analytical off-line measurements.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly[(R)-3-hydroxybutyric acid], natural origin
Sigma-Aldrich
Nile Blue A, certified by the Biological Stain Commission
Sigma-Aldrich
Glycogen from oyster, ≥75% dry basis
Sigma-Aldrich
Glycogen from bovine liver, ≥85%
Sigma-Aldrich
Glycogen from Mytilus edulis (Blue mussel), for DNA precipitations
Sigma-Aldrich
Glycogen from rabbit liver, ≥85% dry basis (enzymatic)
Sigma-Aldrich
Nile Blue A, Dye content ≥75 %
Sigma-Aldrich
Glycogen from Mytilus edulis (Blue mussel), ≥85%
Sigma-Aldrich
Nile Blue chloride, Dye content 85 %
Sigma-Aldrich
Nile Blue A perchlorate, Dye content 95 %
Sigma-Aldrich
Glycogen from oyster, Type XI