2023 journal article
Microplate-Based Cell Viability Assay as a Cost-Effective Alternative to Flow Cytometry for Microalgae Analysis
ENVIRONMENTAL SCIENCE & TECHNOLOGY, 57(50), 21200–21211.
author keywords: cell viability; fluorescent dye; fluoresceindiacetate (FDA); erythrosine B; microplate reader; microalgae; Dunaliella viridis
TL;DR:
This study developed new microplate assays utilizing fluorescein diacetate as a live cell stain and erythrosine B as a dead cell stain that provide a low-cost, simple, and reliable method of assessing cell viability.
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UN Sustainable Development Goal Categories
6. Clean Water and Sanitation
(Web of Science)
Source: Web Of Science
Added: January 2, 2024
Cell viability is a critical indicator for assessing culture quality in microalgae cultivation for biorefinery and bioremediation. Fluorescent dyes that distinguish viable from nonviable cells can enable viability quantification based on the percentage of live cells. However, fluorescence analysis using the typical flow cytometry method is costly and impractical for industrial applications. To address this, we developed new microplate assays utilizing fluorescein diacetate as a live cell stain and erythrosine B as a dead cell stain. These assays provide a low-cost, simple, and reliable method of assessing cell viability. The proposed microplate assays were successfully applied to monitor the viability of the microalgae Dunaliella viridis under carbon and nitrogen limitation stresses and demonstrated good agreement with flow cytometry measurements. We conducted a systematic investigation of the effects of dye concentration, incubation time, and background fluorescence on the microplate assays' performance. Further, we provide a comprehensive review of commonly used fluorescent dyes for microalgae staining, discuss strategies to enhance assay performance, and offer recommendations for dye selection and protocol development. This study presents a comprehensive new method for microplate-based viability analysis, providing valuable insights for future microalgae viability assessments and applications.