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:
||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
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™ 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.
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.
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).