A double-staining automated flow cytometry method for real-time monitoring of bacteria in continuous bioreactors
Year: 2026
Type: article
Abstract: In biotechnological processes, cell density and physiology are critical parameters for controlling the feed rate, harvest time, and process performance. We developed an automated flow cytometry approach that enables continuous, real-time (fully automated, hourly) monitoring of bacterial populations in continuous bioreactors. The method employed a double-staining protocol that combined DAPI to assess total DNA content and Alexa Fluor 488-EdU via Click-iT technology to identify the proportions of cells undergoing active DNA replication through EdU incorporation. The integrated workflow included fixation, permeabilization, staining, and measurement steps and was applied to three Gram-negative strains: Bradyrhizobium sp., Escherichia coli , and Stenotrophomonas rhizophila . Automated analysis captured growth dynamics and cell cycle progression, providing insights into population behavior under different dilution rates. In this study, automated on-line sampling enabled hourly flow cytometry measurements of cell concentration and physiological indicators during continuous cultivation, supporting real-time monitoring and control in industrial biotechnology.
Authors: Juan López-Gálvez, Erik Schönfelder, Hanna Mayer, Konstanze Schiessl, Marisa O. D. Silva, Hauke Harms, Susann Müller
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,490
Awards: G2377581488, HARNESSING THE POWER OF NATURE THROUGH PRODUCTIVE MICROBIAL CONSORTIA IN BIOTECHNOLOGY – MEASURE, MODEL, MASTER, Analyse der lokalen Struktur und Funktion von bakteriellen Kolonien und Biofilmen auf Einzelzellebene mittels Durchflusszytometrie, Zellsortierung, Multiomics und Datenwissenschaft ZPROJECT
OpenAlex ID: W7147147558