Fluorescence-coupled Micropipette Aspiration Assay to Investigate Red Blood Cell Mechanosensing.

Micropipette aspiration assays have long been a cornerstone for the investigation of live-cell mechanics, offering insights into cellular responses to mechanical stress. This paper details an innovative adaptation of the fluorescence-coupled micropipette aspiration (fMPA) assay. The fMPA assay introduces the capability to administer precise mechanical forces while concurrently monitoring the live-cell mechanotransduction processes mediated by ion channels. The sophisticated setup incorporates a precision-engineered borosilicate glass micropipette connected to a finely regulated water reservoir and pneumatic aspiration system, facilitating controlled pressure application with increments as refined as ± 1 mmHg. A significant enhancement is the integration of epi-fluorescence imaging, allowing for the simultaneous observation and quantification of cell morphological changes and intracellular calcium fluxes during aspiration. The fMPA assay, through its synergistic combination of epi-fluorescence imaging with micropipette aspiration, sets a new standard for the study of cell mechanosensing within mechanically challenging environments. This multifaceted approach is adaptable to various experimental setups, providing critical insights into the single-cell mechanosensing mechanisms.

Affective response to volitional input perturbations in obstacle avoidance and target tracking games.

We present the use of two game-like tasks, Catnip and Dinorun, to explore affective responses to volitional control perturbations. We analyze behavioral and physiological measures with the self-assessment manikin (SAM), pupillometry, and electroencephalography (EEG) responses to provide intratrial emotional state as well as inter-trial correlates with selfreported survey responses. We find that subject gameplay characteristics significantly correlate with valence and dominance scores for both games, and that perturbations to the games produce a measurable decrease in response scores for Dinorun. During perturbation events, pupillometry analysis reveals considerable SAM-agnostic dilation, with stronger responses in more rigid trialized event structures. Furthermore, analyses of neural activity from central and parietal regions demonstrate significant measurable evoked responses to perturbed events across the majority of subjects for both games. By introducing perturbations, this set of experiments and analyses inform and enable further studies of affective responses to the loss of volitional control during engaging, game-like tasks.