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DOE OSTI · 2204514

A high-throughput and low-waste viability assay for microbes

Meyer, Christian T. (ORCID:0000000287196522)·Lynch, Grace K. (ORCID:0009000952355456)·Stamo, Dana F.·Miller, Eugene J.·Chatterjee, Anushree (ORCID:0000000283899917)·Kralj, Joel M. (ORCID:0000000193702324)
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Abstract

Abstract Counting viable cells is a universal practice in microbiology. The colony-forming unit (CFU) assay has remained the gold standard to measure viability across disciplines, but it is time-intensive and resource-consuming. Here we describe the geometric viability assay (GVA) that replicates CFU measurements over 6 orders of magnitude while reducing over 10-fold the time and consumables required. GVA computes a sample’s viable cell count on the basis of the distribution of embedded colonies growing inside a pipette tip. GVA is compatible with Gram-positive and Gram-negative planktonic bacteria ( Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis ), biofilms and fungi ( Saccharomyces cerevisiae ). Laborious CFU experiments such as checkerboard assays, treatment time-courses and drug screens against slow-growing cells are simplified by GVA. The ease and low cost of GVA evinces that it can replace existing viability assays and enable viability measurements at previously impractical scales.

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59 BASIC BIOLOGICAL SCIENCESMicrobiologyMolecular biology

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BibTeXRIS

Meyer, Christian T. (ORCID:0000000287196522), Lynch, Grace K. (ORCID:0009000952355456), Stamo, Dana F., Miller, Eugene J., Chatterjee, Anushree (ORCID:0000000283899917), Kralj, Joel M. (ORCID:0000000193702324). 2023-11-02. A high-throughput and low-waste viability assay for microbes. https://doi.org/10.1038/s41564-023-01513-9

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