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Measurement Quality Metrics to Improve Absolute Microbial Cell Counting

Published by National Institute of Standards and Technology | National Institute of Standards and Technology | Metadata Last Checked: August 02, 2025 | Last Modified: 2024-07-15 00:00:00
This repository contains the raw data and analysis scripts supporting the associated publication which introduces a framework to help researchers select fit-for-purpose microbial cell counting methods and optimize protocols for quantification of microbial total cells and viable cells. Escherichia coli cells were enumerated using four methods (colony forming unit assay, impedance flow cytometry - Multisizer 4, impedance flow cytometry - BactoBox, and fluorescent flow cytometry - CytoFLEX LX) and repeated on multiple dates. The experimental design for a single date starts with a cell stock that is divided into 18 sample replicates (3 each for 6 different dilution factors), and each sample is assayed one or two times for a total of 30 observations. Raw data files are provided from the Multisizer 4 (*.#m4) and CytoFLEX LX (*.fcs 3.0). The colony forming unit assay and BactoBox readings are recorded for each date as are the derived results from the Multisizer 4 and CytoFLEX LX. Also provided are an example analysis script for the *.fcs files and the statistical analysis that was performed.

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