Adherent Nucleofection

Dipping Electrode Array - Close up

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  • Introduction

    Electroporation-based methods have so far required cells to be in suspension for transfection. The Nucleofector™ Technology now enters a new era and allows direct Nucleofection of cells, e.g. neurons, in adherence. Adherent Nucleofection can be performed using the the 4D-Nucleofector™ Y Unit:

     Dipping electrode array Conductive polymer Dipping Electrode Arrays

    • Can be inserted into standard 24-well culture plates for Nucleofection
    • Suited for analysis by confocal microscopy or high cell numbers
    • Disposable

    Kits for the 4D-Nucleofector™ Y Unit

    Following our new simplified kit strategy invented with the 4D-Nucleofector™ System we offer two Nucleofector™ Solutions called AD1 (V4YP-1A24) and AD2 (V4YP-2A24) both available as separate kits or combined in an Optimization Kit (V4YP-9A48) . Each solution may serve different cell types. You can easily find out which solution is optimal for your cell of interest by using the following guideline:

    Adherent Nucleofection Scheme

  • Adherent Nucleofection of Neural Cells

    • Nucleofection of neuronal networks at a later developmental stages
    • High transfection efficiencies and viabilities


    For adherent Nucleofection of primary neurons using the 4D-Nucleofector™ Y Unit we a provide basic protocol that helps you to find optimal Nucleofection Conditions for your neuron type of interest. To find out which 4D-Nucleofector™ Kit to order for your neuron type please refer to the cell type specific Nucleofector™ Kit pages or our cell database.

    Mouse cortical neurons - Nucleofection™ in adherence (Y unit) after 6DIV

    Efficient Nucleofection of Neurons in Adherence. Mouse cortical neurons were isolated and seeded into poly-D-lysine coated 24-well plates at a density of 1 x 105 cells/well. After 6 days, cells were transfected with 17.5 μg pmaxGFP™ Vector using the AD1 4D-Nucleofector™ Y Kit and program ER-137. One day post Nucleofection, cells were stained by MAP2 antibody (red) and analyzed by fluorescence microscopy for maxGFP™ protein expression.

  • Adherent Nucleofection of Non-Neural Cells (Y Unit)

    The adherent Nucleofection Mode was originally launched for primary neural cells, but is now also available for non-neuronal cells.


    • More convenient workflow by decoupling transfection from trypsinization
    • Cell can be transfected by Nucleofection at a defined developmental stage

    We provide basic protocols that help you to find optimal Nucleofection Conditions for your cell type of interest.

    • Basic protocol for primary endothelial cells (pdf)
    • General optimization protocol for other primary cells or cell lines of non-neural or non-endothelial origin (pdf)

    To find out which adherent 4D-Nucleofector™ Kit to order for your cell type please refer to the cell type specific Nucleofector™ Kit pages or our cell database.

    HUVEC - Transfected in adherence (Y Unit)  PAEC - Transfected in adherence (Y Unit)

    Human umbilical vein endothelial cells (HUVEC; left) or porcine aortic endothelial cells (PAEC; right) were isolated and plated in passage 1 (HUVEC) or 2 (PAEC) into collagen-coated 24-well plates at a density of 50,000 cells/well. After 1 DIV cells were transfected with 16 µg pmaxGFP™ Vector using AD1 4D-Nucleofector™ Y Solution and program CA-215 (HUVEC) or EW-166 (PAEC). Cells were analyzed for maxGFP™ Protein Expression after 24h. (Data kindly provided by M. Sauvage, Pharmaceutical Industry, FR).