Citation
- Authors: Karimian T. et al.
- Year: 2022
- Journal: Biosensors 12 140
- Applications: in vitro / DNA / jetOPTIMUS
- Cell type: HeLa
Description: Human cervix epitheloid carcinoma cells
Method
HeLa cells were obtained from ATCC, cultured in RPMI medium supplemented with 10% FBS and 1% penicillin/streptomycin and grown at 37 °C in a humidified incubator with 5% CO2.
Cells were transiently transfected with fluorescent-fusion constructs using jetOPTIMUS DNA transfection reagent (Polyplus transfection, Illkirch, France), according to the manufacturer’s protocol.
A total of 24–48 h after transfection, cells were used for subcellular micropatterning experiments.
Abstract
Depositing biomolecule micropatterns on solid substrates via microcontact printing (µCP) usually requires complex chemical substrate modifications to initially create reactive surface groups. Here, we present a simplified activation procedure for untreated solid substrates based on a commercial polymer metal ion coating (AnteoBindTM Biosensor reagent) that allows for direct µCP and the strong attachment of proteins via avidity binding. In proof-of-concept experiments, we identified the optimum working concentrations of the surface coating, characterized the specificity of protein binding and demonstrated the suitability of this approach by subcellular micropatterning experiments in living cells. Altogether, this method represents a significant enhancement and simplification of existing µCP procedures and further increases the accessibility of protein micropatterning for cell biological research questions.