N-acetylgalactosamine (beta-linked, terminal), Vicia villosa lectin, fluorescein conjugated, Vector Laboratories, FL-1231

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  • Single staining with Fluorescein labeled Vicia Villosa Lectin (VVA).

    Antibody: Fluorescein labeled Vicia Villosa Lectin (VVL, VVA) [Vector Labs, Cat: FL-1231], 5, 20 and 35 µg/ml.

    Protocol in short:
    In short, sections were washed (3x10min), treated with target retrieval solution and washed again (3×10 min) Sections were then incubated for 2 hours with fluorescently (488) labeled VVA lectin which has been shown to predominantly stain PV-expressing interneurons. Sections were then again washed (3×10 min) and (1×10 min) in DAPI. Finally, the sections were mounted on slides, air-dried and then mounted with cover slips.

    I find that the VVA specifically stains cells with an acceptable background staining level. Whereas GABA and PV are both localized to the cytoplasm, VVA are distributed along the outer membrane of the cells and thus surrounds the cell bodies. From the 3 concentrations, I have chosen to continue with the 20 µg/ml because the cells were easier to quantify in the microscope with least possible background.

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