A Surface Acoustic Wave (SAW)-Based Lab-on-Chip for the Detection of Active α-Glycosidase.

Enzyme detection in liquid samples is a complex laboratory procedure, based on assays that are generally time- and cost-consuming, and require specialized personnel. Surface acoustic wave sensors can be used for this application, overcoming the cited limitations. To give our contribution, in this work we present the bottom-up development of a surface acoustic wave biosensor to detect active α-glycosidase in aqueous solutions. Our device, optimized to work at an ultra-high frequency (around 740 MHz), is functionalized with a newly synthesized probe 7-mercapto-1-eptyl-D-maltoside, bringing one maltoside terminal moiety. The probe is designed ad hoc for this application and tested in-cuvette to analyze the enzymatic conversion kinetics at different times, temperatures and enzyme concentrations. Preliminary data are used to optimize the detection protocol with the SAW device. In around 60 min, the SAW device is able to detect the enzymatic conversion of the maltoside unit into glucose in the presence of the active enzyme. We obtained successful α-glycosidase detection in the concentration range 0.15-150 U/mL, with an increasing signal in the range up to 15 U/mL. We also checked the sensor performance in the presence of an enzyme inhibitor as a control test, with a signal decrease of 80% in the presence of the inhibitor. The results demonstrate the synergic effect of our SAW Lab-on-a-Chip and probe design as a valid alternative to conventional laboratory tests.

Nanolithographic templates using diblock copolymer films on chemically heterogeneous substrates.

The orientation of the lamellae formed by the phase separation of symmetric diblock copolymer thin films is strongly affected by the wetting properties of the polymer blocks with respect to the substrate. On bare silicon wafers the lamellae of polystyrene-b-polymethylmethacrylate thin films tend to order parallel to the wafer surface, with the polymethylmethacrylate block preferentially wetting silicon. We have developed a methodology for inducing the arrangement of lamellae perpendicular to the substrate by using chemically modified substrates. This is done by chemisorbing a self-assembled monolayer of thiol-terminated alkane chains on thin gold films deposited on silicon wafers. We also show that it is possible to spatially control the perpendicular orientation of the lamellae at sub-micron length scales by using simple chemical patterns and etch them, in order to produce nanolithographic templates. This method may be of great technological interest for the preparation of well-defined templates using block copolymer thin films.

Vertical arrays of nanofluidic channels fabricated without nanolithography.

Vertical arrays of sealed nanofluidic channels, in which both cross-sectional dimensions are controllable down to 10 nm, were fabricated by selective side etching of a SiGe heterostructure comprised of layers of alternating Ge fractions. Capillary filling of these nanochannel arrays with fluorescent dye solutions was investigated using a confocal microscope. The feasibility of using nanochannels for size-based separation of biomolecules was demonstrated by imaging aggregates of tagged amyloid-beta peptide. The ability to integrate a large number of nanochannels shows promise for high throughput applications involving lab-on-a-chip systems.