1. 1 Department of Chemistry; 2 Center for Nanohybrid Functional Materials; 3 Department of Biological Systems Engineering; 4 Biomedical Engineering Program; 5 Department of Chemical and Biomolecular Engineering; 6 Department of Electrical Engineering; University of Nebraska-Lincoln, U.S.A. Characterization of biological thin films with combinatorial spectroscopic ellipsometry and piezoelectric nanogravimetry Biofunctionalizing Sculptured Thin Films Abstract J. Gerasimov 1,2 * K.B. Rodenhausen 2,5, T. Kasputis 3,4, D. Schmidt 2,6, H. Wang 2,6, A.K. Pannier 3, R. Lai 1,2, and M. Schubert 2,6 J. Gerasimov 1,2 * K.B. Rodenhausen 2,5, T. Kasputis 3,4, D. Schmidt 2,6, H. Wang 2,6, A.K. Pannier 3, R. Lai 1,2, and M. Schubert 2,6 Sequence-specific DNA Detection Experimental Design Future Work Acknowledgements: The Procter & Gamble Co., J.A. Woollam Co., Inc., NSF EPS-1004094, NSF CAREER CHE-0955439Nebraska EPSCoR RII External Reviewer Panel Visit, University of Nebraska-Lincoln, August 2011 The formation and function of biotechnologically relevant functional thin films was studied in real-time using tandem spectroscopic ellipsometry (SE) and quartz crystal microbalance (QCM). Simultaneously monitoring changes in film thickness given by each technique allows us to derive the porosity of the bioactive layer under investigation and gives clues about the morphology of the surface-confined biomolecules. A correlation of structural morphology with film function makes possible the directed optimization of bioactive films based on measurable parameters. We present the results of two investigations involving the formation of bioactive hybrid materials. In the study of DNA-incorporating monolayers for sequence-specific DNA detection, two DNA probes were analyzed for their ability to specifically recognize their complement in solution. The formation of protein layers on smooth and nanostructured surfaces was studied for its utility as a scaffold for whole-cell immobilization and future work in gene transfer studies. S SS SSSSSS S SS SSSSS S Probe Adsorption light source polarizer optical window liquid inlet liquid outlet optical window detector QCM-D control gasket surface analyzers polarized light polarized light affected by sample Instrument S SS SSSSS S Target Detection aligned buffer complement target non- complement DNA buffer S SS SSSSS S Electrochemical DNA Sensor Flush Probe Elevated Probe eT Substrate Structured Thin Film Titanium Nanocolumns on Gold extracellular matrix protein facilitates cell adhesion, spreading, and proliferation diverse applications in biotechnology and engineering of hybrid materials Fibronectin Electron transfer (eT) to the labeled, surface-bound stem-loop DNA probe is greatly impeded upon hybridization with complementary target. Decrease in eT efficiency is monitored using alternating current voltammetry SE/QCM provides structural information about DNA layer formation d SE d QCM unbound bound d SE d QCM 12 10 8 6 4 2 0 1.0 0.8 0.6 0.4 0.2 0 120 100 8060 4020 0 d SE d QCM f0vf0v Flat Surface Nanostructured Surface Theory buffer complementary target non- complementary target Study surface functionalization for cell adhesion and gene transfer º Helmholtz Association of German Research Centres (2009, April 7). Fitting Pieces For Biosensors. ScienceDaily. Retrieved August 7, 2011, from http://www.sciencedaily.com­ /releases/2009/04/090407105149.htm ººadapted from http://webs.wichita.edu/mschneegurt/biol103/lecture16/normal_rogue.gif Monitor conformational change in surface-confined proteinsºº In the near future, we will add an electrochemistry module to the SE/QCM to study the relationship between electrochemical trends and film morphology. We will direct further efforts toward the following goals: Characterize peptide-based sensors for antibody detection S SS SSSSS S S SS SSSSS S Aptamer-based small molecule detectionº *jennifer.gerasimov@huskers.unl.edu http://ellipsometry.unl.edu