Conclusions

In this work, the reusability of PAH-modified capacitive field-effect EIS sensors for the label-free electrical detection of ssDNA, in-solution- and on-chip-hybridized dsDNA has been investigated. For this, the formation of five bilayers of PAH/ssDNA or PAH/dsDNA as well as five triple layers of PAH/ssDNA-cDNA onto the EIS-gate surface was monitored by means of dynamic ConCap measurements. It has been demonstrated that via simple regeneration of the EIS-sensor surface by means of adsorption of a new PAH layer, the same biosensor could be reused for at least five DNA-detection measurements. The consecutive adsorption of oppositely charged PAH/ssDNA-, PAH/dsDNA- and PAH/ssDNA-cDNA layers leads to alternating shifts of the ConCap signal. The direction of the EIS-signal shifts depends on the charge sign of the outermost molecular layer and therefore, can be used as an indicator for the verification of successful DNA immobilization or hybridization processes.

In addition, an influence of the Debye-screening effect (which is considered as one of the important factors affecting the sensitivity of FEDs to the macromolecular charge) on the EIS signal has been studied by recording ConCap responses after surface-modification steps in buffer solutions with different ionic strength. The ConCap-signal changes induced by each modification step (i.e., PAH adsorption, immobilization of ssDNA or dsDNA

106 molecules and on-chip hybridization of cDNA) is increased with decreasing the ionic strength of the solution, due to the less efficient screening of the molecular charge of the PAH or DNA by counterions. The results of field-effect measurements were supported by fluorescence-microscopy experiments using PAH- and ssDNA molecules labeled with fluorescence dyes of FITC and FAM, respectively, as well as via staining of the in-solution- and on-chip-hybridized dsDNA with SG dye.

It is worth to note, although in this work, a multilayer PAH/DNA system has been studied, the capacitive EIS platform can be extended for the label-free electrical monitoring of formation of multilayers composed of other oppositely charged cationic/anionic macromolecular systems as well as charged nanoparticle/molecule inorganic/organic nanohybrids.

A

CKNOWLEDGEMENTS

The authors thank S. Scheja for technical support, H. Iken for wafer processing and C.

Metzger-Boddien, H. Busch for valuable discussions. The authors wish to acknowledge the German Federal Ministry of Education and Research for the financial support (project

“DiaCharge”, grant no.: 031A192D), Germany.

107

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