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

Sandra P Mai-Morente; Juan P Irigoyen; Victoria M Carriquiry; Virginia M Marset; Mariana Di Doménico; Eugenia Isasi; Verónica Abudara

doi:10.21769/BioProtoc.4222

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.

Authors

Sandra P Mai-Morente¹, Juan P Irigoyen¹, Victoria M Carriquiry¹, Virginia M Marset¹, Mariana Di Doménico², Eugenia Isasi³, Verónica Abudara¹

Affiliations

¹ Departamento de Fisiología, Facultad de Medicina, Universidad de la República, General Flores 2125, Montevideo, CP 11 800, Uruguay.
² Departamento de Biofísica, Facultad de Medicina, Universidad de la República, General Flores 2125, Montevideo, CP 11 800, Uruguay.
³ Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, General Flores 2125, Montevideo, CP 11 800, Uruguay.

Abstract

This protocol describes a method for high-resolution confocal imaging of pericytes with the far-red fluorophore TO-PRO^TM-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.