EasyEyes - A new method for accurate fixation in online vision testing.

Online methods allow testing of larger, more diverse populations, with much less effort than in-lab testing. However, many psychophysical measurements, including visual crowding, require accurate eye fixation, which is classically achieved by testing only experienced observers who have learned to fixate reliably, or by using a gaze tracker to restrict testing to moments when fixation is accurate. Alas, both approaches are impractical online as online observers tend to be inexperienced, and online gaze tracking, using the built-in webcam, has a low precision (±4 deg). EasyEyes open-source software reliably measures peripheral thresholds online with accurate fixation achieved in a novel way, without gaze tracking. It tells observers to use the cursor to track a moving crosshair. At a random time during successful tracking, a brief target is presented in the periphery. The observer responds by identifying the target. To evaluate EasyEyes fixation accuracy and thresholds, we tested 12 naive observers in three ways in a counterbalanced order: first, in the laboratory, using gaze-contingent stimulus presentation; second, in the laboratory, using EasyEyes while independently monitoring gaze using EyeLink 1000; third, online at home, using EasyEyes. We find that crowding thresholds are consistent and individual differences are conserved. The small root mean square (RMS) fixation error (0.6 deg) during target presentation eliminates the need for gaze tracking. Thus, this method enables fixation-dependent measurements online, for easy testing of larger and more diverse populations.

EasyEyes - Accurate fixation for online vision testing of crowding and beyond.

Online methods allow testing of larger, more diverse populations, with much less effort than in-lab testing. However, many psychophysical measurements, including visual crowding, require accurate eye fixation, which is classically achieved by testing only experienced observers who have learned to fixate reliably, or by using a gaze tracker to restrict testing to moments when fixation is accurate. Alas, both approaches are impractical online since online observers tend to be inexperienced, and online gaze tracking, using the built-in webcam, has a low precision (±4 deg, Papoutsaki et al., 2016). The EasyEyes open-source software reliably measures peripheral thresholds online with accurate fixation achieved in a novel way, without gaze tracking. EasyEyes tells observers to use the cursor to track a moving crosshair. At a random time during successful tracking, a brief target is presented in the periphery. The observer responds by identifying the target. To evaluate EasyEyes fixation accuracy and thresholds, we tested 12 naive observers in three ways in a counterbalanced order: first, in the lab, using gaze-contingent stimulus presentation (Kurzawski et al., 2023; Pelli et al., 2016); second, in the lab, using EasyEyes while independently monitoring gaze; third, online at home, using EasyEyes. We find that crowding thresholds are consistent (no significant differences in mean and variance of thresholds across ways) and individual differences are conserved. The small root mean square (RMS) fixation error (0.6 deg) during target presentation eliminates the need for gaze tracking. Thus, EasyEyes enables fixation-dependent measurements online, for easy testing of larger and more diverse populations.

Propagation dynamics of electrotactic motility in large epithelial cell sheets.

Directional migration initiated at the wound edge leads epithelia to migrate in wound healing. How such coherent migration is achieved is not well understood. Here, we used electric fields to induce robust migration of sheets of human keratinocytes and developed an in silico model to characterize initiation and propagation of epithelial collective migration. Electric fields initiate an increase in migration directionality and speed at the leading edge. The increases propagate across the epithelial sheets, resulting in directional migration of cell sheets as coherent units. Both the experimental and in silico models demonstrated vector-like integration of the electric and default directional cues at free edge in space and time. The resultant collective migration is consistent in experiments and modeling, both qualitatively and quantitatively. The keratinocyte model thus faithfully reflects key features of epithelial migration as a coherent tissue in vivo, e.g. that leading cells lead, and that epithelium maintains cell-cell junction.

Collective cell migration has distinct directionality and speed dynamics.

When a constraint is removed, confluent cells migrate directionally into the available space. How the migration directionality and speed increase are initiated at the leading edge and propagate into neighboring cells are not well understood. Using a quantitative visualization technique-Particle Image Velocimetry (PIV)-we revealed that migration directionality and speed had strikingly different dynamics. Migration directionality increases as a wave propagating from the leading edge into the cell sheet, while the increase in cell migration speed is maintained only at the leading edge. The overall directionality steadily increases with time as cells migrate into the cell-free space, but migration speed remains largely the same. A particle-based compass (PBC) model suggests cellular interplay (which depends on cell-cell distance) and migration speed are sufficient to capture the dynamics of migration directionality revealed experimentally. Extracellular Ca2+ regulated both migration speed and directionality, but in a significantly different way, suggested by the correlation between directionality and speed only in some dynamic ranges. Our experimental and modeling results reveal distinct directionality and speed dynamics in collective migration, and these factors can be regulated by extracellular Ca2+ through cellular interplay. Quantitative visualization using PIV and our PBC model thus provide a powerful approach to dissect the mechanisms of collective cell migration.

A compact microfluidic system for cell migration studies.

Microfluidic systems can better control cellular microenvironments and therefore are increasingly used for cell migration research. However, most existing systems are impractical to use without specialized facilities and researchers. Toward removing this barrier, we developed a compact USB microscope-based Microfluidic Chemotaxis Analysis System (UMCAS). This system integrates microfluidic devices, live cell imaging, environmental control and data analysis to provide a solution for rapid microfluidic cell migration and chemotaxis experiments with real-time result reporting. This developed system was successfully validated by testing neutrophil chemotaxis.

Accurate computation of Zernike moments in polar coordinates.

An algorithm for high-precision numerical computation of Zernike moments is presented. The algorithm, based on the introduced polar pixel tiling scheme, does not exhibit the geometric error and numerical integration error which are inherent in conventional methods based on Cartesian coordinates. This yields a dramatic improvement of the Zernike moments accuracy in terms of their reconstruction and invariance properties. The introduced image tiling requires an interpolation algorithm which turns out to be of the second order importance compared to the discretization error. Various comparisons are made between the accuracy of the proposed method and that of commonly used techniques. The results reveal the great advantage of our approach.