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

Abstract

Cellular mechanical properties can reveal physiologically relevant characteristics in many cell types, and several groups have developed microfluidics-based platforms to perform single-cell mechanical testing with high throughput. 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, which is 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, and 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 labs 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.

Neurodesk: an accessible, flexible and portable data analysis environment for reproducible neuroimaging

“…However, this is impractical and too time-consuming to implement at review 8 . Where analysis pipelines are ported, subtle differences in the implementation of specific processing steps and software versions across computing environments can systematically affect results [21][22][23][24] . Thus, it is often impossible to reproduce a prior study's results, even given the original data and analysis protocol 14,21 .…”

Neurodesk: An accessible, flexible, and portable data analysis environment for reproducible neuroimaging

“…However, this is impractical and too time-consuming to implement at review 8 . Where analysis pipelines are ported, subtle differences in the implementation of specific processing steps and so ware versions across computing environments can systematically affect results [21][22][23][24] . Thus, it is o en impossible to reproduce a prior study's results, even given the original data and analysis protocol 14,21 .…”