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Biology subjects

Gillette, M.

Publications and source records attributed to Gillette, M..

2 recordsLinked to original sources

Optical excitation and detection of neuronal activity

Optogenetics has emerged as an exciting tool for manipulating neural activity, which in turn, can modulate behavior in live organisms. However, detecting the response to the optical stimulation requires electrophysiology with physical contact or fluorescent imaging at target locations, which is often limited by photobleaching and phototoxicity. In this paper, we show that phase imaging can report the intracellular transport induced by optogenetic stimulation. We developed a multimodal instrument that can both stimulate cells with subcellular spatial resolution and detect optical pathlength changes with nanometer scale sensitivity. We found that optical pathlength fluctuations following stimulation are consistent with active organelle transport. Furthermore, the results indicate a broadening in the transport velocity distribution, which is significantly higher in stimulated cells compared to optogenetically inactive cells. It is likely that this label-free, contactless measurement of optogenetic response will provide an enabling approach to neuroscience.\n\nO_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY Quantitative phase imaging enables label-free detection of enhanced intracellular activities induced by optogenetic stimulation.\n\nC_FIG_DISPLAY

bioengineering

Less is More: Oligomer extraction and hydrothermal annealing increase PDMS bonding forces for new microfluidics assembly and for biological studies

Key determinants in the emergence of complex cellular morphologies and functions are cues in the micro-environment. Primary among these is the presence of neighboring cells as networks form. Therefore, for high-resolution analysis, it is crucial to develop micro-environments that permit exquisite control of network formation. This is especially true in cell science, tissue engineering, and clinical biology. We introduce a new approach for assembling polydimethylsiloxane (PDMS)-based microfluidic environments that enhances cell network formation and analyses. We report that the combined processes of PDMS solvent-extraction (E-PDMS) and hydrothermal annealing create unique conditions that produce high-strength bonds between E-PDMS and glass - properties not associated with conventional PDMS. Extraction followed by hydrothermal annealing removes unbound oligomers, promotes polymer cross-linking, facilitates covalent bond formation with glass, and retains the highest biocompatibility. Our extraction protocol accelerates oligomer removal from 5 to 2 days. Resulting microfluidic platforms are uniquely suited for cell-network studies owing to high bond strengths, effectively corralling cellular extensions and eliminating harmful oligomers. We demonstrate simple, simultaneous actuation of multiple microfluidic domains for invoking ATP- and glutamate-induced Ca2+ signaling in glial-cell networks. These low-cost, simple E-PMDS modifications and flow manipulations further enable microfluidic technologies for cell-signaling and network studies as well as novel applications.

bioengineering