Investigations of Astrocyte Calcium Signaling and Imaging with Classical and Nonclassical Light.

The application of light in studying and influencing cellular behavior with improved temporal and spatial resolution remains a key objective in fields such as chemistry, physics, medicine, and engineering. In the brain, nonexcitable cells called astrocytes play essential roles in regulating homeostasis and cognitive function through complex calcium signaling pathways. Understanding these pathways is vital for deciphering brain physiology and neurological disorders like Parkinson's and Alzheimer's. Despite challenges in selectively targeting astrocyte signaling pathways due to shared molecular equipment with neurons, recent advancements in laser technology offer promising avenues. However, the effort to use laser light properties to study astroglial cell function is still limited. This work aims to exploit an in-depth pharmacological analysis of astrocyte calcium channels to determine the physiological mechanism induced by exposure to classical nanosecond-pulsed light. We herein report molecular clues supporting the use of visible-nanosecond laser pulses as a promising approach to excite primary rat neocortical astrocytes and unprecedentedly report on the implementation of entangled two-photon microscopy to image them.