Pericyte Mapping in Cerebral Slices with the Far-red Fluorophore TO-PRO-3

Bio Protoc. 2021 Nov 20;11(22):e4222. doi: 10.21769/BioProtoc.4222.


This protocol describes a method for high-resolution confocal imaging of pericytes with the far-red fluorophore TO-PROTM-3 Iodide 642/661 in cerebral slices of murine. Identification of pericytes with TO-PRO-3 is a short time-consuming, high cost-effective and robust technique to label pericytes with no need for immunostaining or generation of reporter mice. Since the TO-PRO-3 stain resists immunofluorescence, and lacks spectral overlap, the probe is well suited for multiple labelling. Our procedures also combine TO-PRO-3-staining of pericytes with fluorescent markers for astrocytes and vessels in brain slices. These approaches should enable the assessment of pericyte biology in gliovascular unit.

Keywords: Astrocytes; Brain slices; Fluorescence confocal microscopy; Pericyte imaging; TO-PRO-3; vessels.