Computational protocol: Revisiting the Lamotrigine-Mediated Effect on Hippocampal GABAergic Transmission

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Protocol publication

[…] Neurons were filled with biocytin (2 mg/mL) during whole-cell recordings and subsequently fixed overnight with 4% paraformaldehyde in phosphate-buffered solution (PB; 0.1 M, pH 7.3). After washing with PB, slices were incubated with fluorescein isothiocyanate-conjugated avidin-d (2 µL/mL; Invitrogen, Eugene, OR, USA) in PB and 0.3% Triton X-100 overnight at 4 °C. After washing, slices were embedded in mounting medium Vectashield® (Vector Laboratories, Burlingame, CA, USA). Labeled cells were imaged by a two-photon microscope as described before []. The two-dimensional morphology of the cells was reconstructed from a stack of 100 images (voxel size, 0.378–1.514 μm in the x–y plane; 0.4–0.99 μm along the z-axis) using ImageJ (v.1.42q) or Neuromantic 1.6.3 software. […]

Pipeline specifications

Software tools ImageJ, Neuromantic
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Rattus norvegicus
Chemicals gamma-Aminobutyric Acid, Nucleotides, Cyclic, Sodium