Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping

• Published in: PloS one Vol. 16; no. 10; p. e0258982
• Main Authors: Li, Brian, Cotner, Kristen L., Liu, Nathaniel K., Hinz, Stefan, LaBarge, Mark A., Sohn, Lydia L.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.10.2021; Public Library of Science (PLoS)
• Subjects: Analysis; Bioengineering; Biology and Life Sciences; Biosensors; Cell physiology; Computer and Information Sciences; Data processing; Deformation; Design; Engineering and Technology; Flow Cytometry; Laboratory tests; Mechanical engineering; Mechanical measurement; Mechanical properties; Mechanical tests; Medicine and Health Sciences; Microfluidics; Microfluidics - methods; Nodes; Performance evaluation; Phenotype; Phenotyping; Platforms; Reproducibility; Reproducibility of Results; Research and Analysis Methods; Single-Cell Analysis; Social Sciences; Trends; Variability; Viscoelasticity; United States; United States > US; San Francisco California; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0258982

Cellular mechanical properties can reveal physiologically relevant characteristics in many cell types, and several groups have developed microfluidics-based platforms to perform high-throughput single-cell mechanical testing. However, prior work has performed only limited characterization of these platforms’ technical variability and reproducibility. Here, we evaluate the repeatability performance of mechano-node-pore sensing, a single-cell mechanical phenotyping platform developed by our research group. We measured the degree to which device-to-device variability and semi-manual data processing affected this platform’s measurements of single-cell mechanical properties. We demonstrated high repeatability across the entire technology pipeline even for novice users. We then compared results from identical mechano-node-pore sensing experiments performed by researchers in two different laboratories with different analytical instruments, demonstrating that the mechanical testing results from these two locations are in agreement. Our findings quantify the expectation of technical variability in mechano-node-pore sensing even in minimally experienced hands. Most importantly, we find that the repeatability performance we measured is fully sufficient for interpreting biologically relevant single-cell mechanical measurements with high confidence.