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 in two different ways:
- Nucleocuvette™ AD strips in rack
4D-Nucleofector™ X unit (or 96-well Shuttle™ add-on)
- Utilizes specialized 16-well Nucleocuvette™ AD strips (20 μl) in which neural cells can be cultured for up to 14 days and transfected at any time during this period
- Suited for applications requiring low cell numbers analyzed by fluorescence microscopy, absorption, fluorescence or luminescence assays
- Dipping electrode array
4D-Nucleofector™ Y unit
- Works with disposable conductive polymer dipping electrode arrays that can be inserted into standard 24-well culture plates for the Nucleofection™
- Suited for analysis by confocal microscopy or high cell numbers
Adherent Nucleofection™ formats
| Nucleocuvette™ AD strips (20 µl) |
Dipping electrode arrays | |
| Required functional 4D-Nucleofector™ unit | X unit or 96-well Shuttle™ |
Y unit |
| Pre- and post Nucleofection™ culture | Nucleocuvette™ wells |
24-well culture plate |
| Nucleofection™ of cells plated on glass cover slips | – |
• |
| High transfection efficiencies and viabilities | • | • |
| Cell analysis by: – transmission light or fluorescence microscopy |
• | • |
| – absorption, luminescence or fluroescence assays | • | • |
| – confocal microscopy | – | • |
| – patch clamping | – | • |
| Nucleofection™ of cryopreserved Clonetics™ primary animal neurons | • | • |
Adherent Nucleofection™ of neurons
For adherent Nucleofection™ of primary neurons using the different Nucleofector™ systems we provide basic protocols that help you to find optimal Nucleofection™ conditions for your neuron type of interest:
- Nucleofection™ of neuronal networks at a later developmental stages
- High transfection efficiencies and viabilities
To find out which 4D- or 96-well Nucleofector™ kit to order for your neuron type please refer to our e-shop or 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.
To request a demonstration for neurons, receive more information or ask for further upcoming adherent Nucleofector™ Kits
