Rapid quantification of intracellular polyhydroxyalkanoates via fluorescence techniques: A critical review

Abstract

Polyhydroxyalkanoates (PHA) are promising bioplastics with excellent physicochemical properties and biodegradability, whereas PHA products suffer from high manufacturing costs. To reduce costs of PHA production, experiments with mixed microbial cultures and low-cost substrates have been conducted widely, where rapid and robust PHA quantification methods are necessary. Compared with traditional gas chromatography methods, PHA fluorescence quantification (PHA-FQ) methods may be quicker, safer and more suitable for modern experiments with high throughput requirements. However, practical applications of PHA-FQ methods are still limited. Therefore, this review provides a comprehensive understanding of PHA-FQ methods. Performance of PHA-staining fluorochromes, relevant spectral properties, and important staining procedures are summarized. Current developments of PHA-FQ protocols are critically reviewed. Main considerations needed to make PHA-FQ protocol reliable are comprehensively discussed. Finally, potential improvements in various aspects of PHA-FQ methods are highlighted. This review could help researchers develop more effective PHA-FQ methods and facilitate future experiments related to PHA.

Keywords: Fluorescence quantification; Fluorescent staining; Partial least squares (PLS); Polyhydroxyalkanoates (PHA); Quantitative image analysis (QIA).