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How Simplified Cell Count and Viability Measurements Work

Problem: The assessment of cell concentration and viability is an important step in the characterization of cell health. This information can be used for monitoring proliferation rates, optimizing growth conditions and normalizing cell data for further studies, such as assessing the impacts of cytotoxic compounds. Current methods rely on multiple, sometimes complex, instrument platforms to provide these answers, reducing flexibility and increasing research costs. Other, simpler methods provide inconsistent results due to their dependence on single-uptake dyes, which do not effectively discriminate between the various states of cell demise. As a result, there is a crucial need for analytical methods that efficiently provide reproducible count and viability data.

by EMD Millipore
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The Muse® Cell Analyzer.EMD Millipore

Solution: The Muse® Cell Analyzer from EMD Millipore is a miniaturized flow cytometer that enables measurement of multiple cell health-related parameters on a single, benchtop platform. Using multiparametric fluorescent detection of individual cells via microcapillary flow technology, the system enables highly sensitive and rapid detection of cellular samples using minimal cell numbers.

The accompanying Muse® Count & Viability Assay is a simple, rapid assay that provides cell concentration and viability information. It uses a proprietary mix of two DNA intercalating fluorescent dyes in a single reagent. One of the dyes is membrane-permeant and stains all cells with a nucleus. The second dye only stains cells whose membranes have been compromised and are dying or dead. This combination allows for the discrimination of nucleated cells from those without a nucleus or debris, and live cells from dead or dying, resulting in both accurate cell concentration and viability results. The use of these dual fluorescent probes allows for greater sensitivity and accuracy compared to colorimetric methods.

The assay uses a highly simplified workflow to provide accurate and precise count and viability results. Sample preparation is simple, with the one-step addition of the mix-and-read Muse® Count & Viability reagent, allowing for rapid measurement and instantaneous results. Stained samples are analyzed on the Muse® Cell Analyzer using a guided touchscreen user interface.

The touchscreen prompts the user to load a sample and, through simple on-screen instructions, guides the user through the optimization and verification of settings. The user then enters sample-specific information and touches “Run Sample.” The instrument displays an easy-to-read results screen with the calculated concentration values and provides the option to view the dotplot and adjust markers between samples.

Result parameters include the number of viable cells per milliliter and in the original sample, the total cells per milliliter and the percent viability. Data can be stored on the device, exported in a report format and/or exported as a Microsoft Excel® file, thus enabling the production of a robust documentation trail with experimental details preserved.

The assay requires only small cell sample sizes and has been validated on a variety of both suspension and adherent cell lines. Performance data demonstrate strong correlations with accepted analysis methods, confirm this new platform yields accurate count and viability results for a variety of cell types and concentrations, and demonstrate its excellent precision compared to traditional methods. By making cell count and viability analysis consistent and simple for researchers of all backgrounds, products such as the Muse® Cell Analyzer and accompanying Count & Viability Assay allow for more productive cell health assessment.

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