The increase of the viability of neuroinflammation related glial cells such as astrocytes and microglia by peptide drugs has been proven in Alzheimer’s Disease (AD) mouse models through in vitro experiments. Also, astrocytes and microglia that possess amyloid beta (Aβ) deposits show neuroprotective effects in the early stages of AD but change their function and promote AD development when prolonged with inflammatory responses. However, the exact cellular interactions that occur in these cells from the accumulation of Aβ in vivo remain unclear. Here, in vivo imaging was utilized to monitor the direct effects of a peptide drug to the viabilities of astrocytes and microglia that express a hormone receptor against an Aβ treatment group and their changes in phenotypes. A cranial imaging window insertion was performed on the skulls of 3xTg-AD mice that were treated with the peptide drug followed by longitudinal intravital imaging. Whilst Aβ reduced the viability of the astrocytes and microglia, a significant proportion was restored in the peptide drug induced groups. Furthermore, the peptide drug reduced the expression of neurotoxic markers and increased the expression of neuroprotective markers both in the astrocytes and microglia. In vivo imaging has shed light to the direct interactions that occur on glial cells from the treatment of such peptide drug and thereby bringing new possibilities in monitoring such interactions of other developmental stage drugs on the treatment of AD.