Spectral Algal Fingerprinting and Long Sequencing in Synthetic Algal–Microbial Communities

Abstract

Synthetic biology has advanced in creating artificial microbial and algal communities, but technical and evolutionary complexities still pose significant challenges. Traditional methods, like microscopy and pigment analysis, are limited in throughput and resolution. In contrast, advancements in full-spectrum cytometry enabled high-throughput, multidimensional analysis of single cells based on size, complexity, and spectral fingerprints, offering more precision and flexibility than conventional flow cytometry. This study uses full-spectrum cytometry to analyze synthetic algal–microbial communities, enabling rapid species identification and enumeration. The workflow involves recording individual spectral signatures from monocultures, using autofluorescence to capture populations of interest, and creating a spectral library for further analysis. This spectral library was used for the analysis of the synthetic phytoplankton communities, revealing differences in spectral signatures. Moreover, the synthetic consortium experiment monitored algal growth, comparing results from different instruments, highlighting the advantages of the spectral virtual filter system for precise population separation and abundance tracking. By capturing the entire emission spectrum of each cell, this method enhances understanding of algal–microbial community dynamics and responses to environmental stressors. The development of standardized spectral libraries would improve the characterization of algal communities, further advancing synthetic biology and phytoplankton ecology research.

Keywords

• synthetic algal–microbial communities
• spectral flow cytometry
• imaging flow cytometry
• long sequencing
• nanopore-based sequencing