Abstract
The purpose of the present study is to develop an analytical method for automatic detection of the brain cell activation, imaged with two-photon microscopy in the somatosensory cortex of awake mice. The cells expressed calcium-sensitive genetic sensor (GCaMP3) under neural promoter of CaMKII, were also labeled with sulforhodamine 101 (SR101), a marker of glial cells. The software detected the active cells which showed enhanced fluorescent signal during contralateral whisker stimulation, and measured a location of the cells from the cortical surface with a distance from the nearest blood vessels which were also labeled with SR101. In addition, the cells were further classified into neurons and glial cells based on SR101 staining. The present method will be a useful technique to determine a chronological order of functional plasticity/degeneration at the cellular level based on massive cellular imaging data for learning or a development of neurological disorders, cognitive impairment in the long term changes in neurovascular unit.
