GluN2B Subunit Labeling with Fluorescent Probes and High-Resolution Live Imaging

Methods Mol Biol. 2017:1677:171-183. doi: 10.1007/978-1-4939-7321-7_9.


Laser Scanning Confocal Microscopy (LSCM) imaging using an appropriate fluorescent probe enables the visualization of a molecular target with high resolution, and represents a method of choice for studying expression, subcellular location, and trafficking of receptors in living cells. The chemical, physical, and pharmacological properties of the probe remain essential. Here, we describe (1) the preparation of a specific probe for NMDAR GluN2B receptor by conjugation of fluorescein to an ifenprodil-based ligand, (2) an in vitro functional assay by calcium imaging for GluN2B binding and inhibition evaluation of the probe, and (3) the labeling and confocal imaging of GluN2B in DS-red labeled living cortical neurons.

Keywords: Calcium imaging; Confocal microscopy; Fluorescein; GluN2B; ifenprodil.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / chemistry*
  • Fluorescent Dyes / chemistry
  • Microscopy, Confocal / methods*
  • Piperidines / chemistry
  • Receptors, N-Methyl-D-Aspartate / chemistry*


  • Fluorescent Dyes
  • NR2B NMDA receptor
  • Piperidines
  • Receptors, N-Methyl-D-Aspartate
  • ifenprodil
  • Calcium