Dataset Information

Improving Quality, Reproducibility, and Usability of FRET-Based Tension Sensors.

ABSTRACT: Mechanobiology, the study of how mechanical forces affect cellular behavior, is an emerging field of study that has garnered broad and significant interest. Researchers are currently seeking to better understand how mechanical signals are transmitted, detected, and integrated at a subcellular level. One tool for addressing these questions is a Förster resonance energy transfer (FRET)-based tension sensor, which enables the measurement of molecular-scale forces across proteins based on changes in emitted light. However, the reliability and reproducibility of measurements made with these sensors has not been thoroughly examined. To address these concerns, we developed numerical methods that improve the accuracy of measurements made using sensitized emission-based imaging. To establish that FRET-based tension sensors are versatile tools that provide consistent measurements, we used these methods, and demonstrated that a vinculin tension sensor is unperturbed by cell fixation, permeabilization, and immunolabeling. This suggests FRET-based tension sensors could be coupled with a variety of immuno-fluorescent labeling techniques. Additionally, as tension sensors are frequently employed in complex biological samples where large experimental repeats may be challenging, we examined how sample size affects the uncertainty of FRET measurements. In total, this work establishes guidelines to improve FRET-based tension sensor measurements, validate novel implementations of these sensors, and ensure that results are precise and reproducible. © 2018 International Society for Advancement of Cytometry.

SUBMITTER: Gates EM

PROVIDER: S-EPMC6375766 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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Improving Quality, Reproducibility, and Usability of FRET-Based Tension Sensors.

Gates Evan M EM LaCroix Andrew S AS Rothenberg Katheryn E KE Hoffman Brenton D BD

Cytometry. Part A : the journal of the International Society for Analytical Cytology 20181206 2

Mechanobiology, the study of how mechanical forces affect cellular behavior, is an emerging field of study that has garnered broad and significant interest. Researchers are currently seeking to better understand how mechanical signals are transmitted, detected, and integrated at a subcellular level. One tool for addressing these questions is a Förster resonance energy transfer (FRET)-based tension sensor, which enables the measurement of molecular-scale forces across proteins based on changes in ...[more]

PMID: 30523675

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Project description:BackgroundSymptom drawings are widely used as a qualitative and quantitative method of assessing pain symptoms for both clinical and research purposes. As electronic drawings offer many advantages over classical pen-and-paper drawings, the last years have seen a shift toward tablet-based acquisition of symptom drawings. However, software that is used in clinical care requires special attention to usability aspects and design to provide easy access for physically impaired or elderly patients.ObjectiveThe aims of this project were to develop a new tablet-based software app specifically designed to collect patients' and doctors' drawings of pain and related bodily symptoms and test it for usability in 2 samples of chronic pain patients (Aim 1) and their treating doctors (Aim 2) as well as for test-retest reliability (Aim 3).MethodsIn 2 separate studies, symptom drawings from 103 chronic pain patients and their treating doctors were collected using 2 different versions of the app. Both patients and doctors evaluated usability aspects of the app through questionnaires. Results from study 1 were used to improve certain features of the app, which were then evaluated in study 2. Furthermore, a subgroup of 25 patients in study 2 created 2 consecutive symptom drawings for test-retest reproducibility analysis. Usability of both app versions was compared, and reproducibility was calculated for symptom extent, number of symptom clusters, and the whole symptom pattern.ResultsThe changes we made to the app and the body outline led to significant improvements in patients' usability evaluation regarding the identification with the body outline (P=.007) and the evaluation of symptom depth (P=.02), and the overall difficultness of the drawing process (P=.003) improved significantly. Doctors' usability evaluation of the final app showed good usability with 75.63 (SD 19.51) points on the System Usability Scale, Attrakdiff 2 scores from 0.93 to 1.41, and ISONORM 9241/10 scores from -0.05 to 1.80. Test-retest analysis showed excellent reproducibility for pain extent (intraclass correlation coefficient, ICC=0.92) and good results for the number of symptom clusters (ICC=0.70) and a mean overlap of 0.47 (Jaccard index).ConclusionsWe developed a tablet-based symptom drawing app and improved it based on usability assessment in a sample of chronic pain patients and their treating doctors. Increases in usability of the improved app comprised identification with the body outline, symptom depth evaluation, and difficultness of the drawing process. Test-retest reliability of symptom drawings by chronic pain patients showed fair to excellent reproducibility. Patients' usability evaluation is an important factor that should not be neglected when designing apps for mobile or eHealth apps.

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Dataset Information

Improving Quality, Reproducibility, and Usability of FRET-Based Tension Sensors.

Publications

Improving Quality, Reproducibility, and Usability of FRET-Based Tension Sensors.

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