Studying the mechanism of neurostimulation by infrared laser light using GCaMP6s and Rhodamine B imaging
Résumé
Infrared laser light radiation can be used to depolarize neurons and to stimulate neural
activity. The absorption of infrared radiation and heating of biological tissue is thought to
be the underlying mechanism of this phenomenon whereby local temperature increases in
the plasma membrane of cells either directly influence membrane properties or act via
temperature sensitive ion channels. Action potentials are typically measured electrically in
neurons with microelectrodes, but they can also be observed using fluorescence
microscopy techniques that use synthetic or genetically encoded calcium indicators. In
this work, we studied the impact of infrared laser light on neuronal calcium signals to
address the mechanism of these thermal effects. Cultured primary mouse hippocampal
neurons expressing the genetically encoded calcium indicator GCaMP6s were used in
combination with the temperature sensitive fluorophore Rhodamine B to measure calcium
signals and temperature changes at the cellular level. Here we present our all-optical
strategy for studying the influence of infrared laser light on neuronal activity. © (2016)
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