A high-throughput and low-waste viability assay for microbes.
Christian T MeyerGrace K LynchDana F StamoEugene J MillerAnushree ChatterjeeJoel M KraljPublished in: Nature microbiology (2023)
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.
Keyphrases
- high throughput
- gram negative
- single cell
- induced apoptosis
- multidrug resistant
- saccharomyces cerevisiae
- low cost
- escherichia coli
- pseudomonas aeruginosa
- cell cycle arrest
- bacillus subtilis
- primary care
- cystic fibrosis
- healthcare
- acinetobacter baumannii
- endoplasmic reticulum stress
- cell death
- heavy metals
- gene expression
- peripheral blood
- genome wide
- cell therapy
- signaling pathway
- mesenchymal stem cells
- dna methylation
- drug resistant
- adverse drug
- bone marrow