Michael Bache

High throughput label-free platform for statistical bio-molecular sensing.

Sensors are crucial in many daily operations including security, environmental control, human diagnostics and patient monitoring. Screening and online monitoring require reliable and high-throughput sensing. We report on the demonstration of a high-throughput label-free sensor platform utilizing cantilever based sensors. These sensors have often been acclaimed to facilitate highly parallelized operation. Unfortunately, so far no concept has been presented which offers large datasets as well as easy liquid sample handling. We use optics and mechanics from a DVD player to handle liquid samples and to read-out cantilever deflection and resonant frequency. Also, surface roughness is measured. When combined with cantilever deflection the roughness is discovered to hold valuable additional information on specific and unspecific binding events. In a few minutes, 30 liquid samples can be analyzed in parallel, each by 24 cantilever-based sensors. The approach was used to detect the binding of streptavidin and antibodies.

Nanomechanical recognition of prognostic biomarker suPAR with DVD-ROM optical technology.

In this work the use of a high-throughput nanomechanical detection system based on a DVD-ROM optical drive and cantilever sensors is presented for the detection of urokinase plasminogen activator receptor inflammatory biomarker (uPAR). Several large scale studies have linked elevated levels of soluble uPAR (suPAR) to infectious diseases, such as HIV, and certain types of cancer. Using hundreds of cantilevers and a DVD-based platform, cantilever deflection response from antibody-antigen recognition is investigated as a function of suPAR concentration. The goal is to provide a cheap and portable detection platform which can carry valuable prognostic information. In order to optimize the cantilever response the antibody immobilization and unspecific binding are initially characterized using quartz crystal microbalance technology. Also, the choice of antibody is explored in order to generate the largest surface stress on the cantilevers, thus increasing the signal. Using optimized experimental conditions the lowest detectable suPAR concentration is currently around 5 nM. The results reveal promising research strategies for the implementation of specific biochemical assays in a portable and high-throughput microsensor-based detection platform.

Investigations on antibody binding to a micro-cantilever coated with a BAM pesticide residue

The attachment of an antibody to an antigen-coated cantilever has been investigated by repeated experiments, using a cantilever-based detection system by Cantion A/S. The stress induced by the binding of a pesticide residue BAM (2,6 dichlorobenzamide) immobilized on a cantilever surface to anti-BAM antibody is measured using the CantiLab4© system from Cantion A/S with four gold-coated cantilevers and piezo resistive readout. The detection mechanism is in principle label-free, but fluorescent-marked antibodies have been used to subsequently verify the binding on the cantilever surface. The bending and increase in mass of each cantilever has also been investigated using a light interferometer and a Doppler Vibrometer. The system has been analyzed during repeated measurements to investigate whether the CantiLab4© system is a suited platform for a pesticide assay system.

High-throughput automated system for statistical biosensing employing microcantilever arrays

In this paper we present a completely new and fully automated system for parallel microcantilever-based biosensing. Our platform is able to monitor simultaneously the change of resonance frequency (dynamic mode), of deflection (static mode), and of surface roughness of hundreds of cantilevers in a very short time over multiple biochemical reactions. We have proven that our system is capable to measure 900 independent microsensors in less than a second. Here, we report statistical biosensing results performed over a haptens-antibody assay, where complete characterization of the biochemical binding on the cantilever surfaces is obtained with higher accuracy than standard optical lever-based setups.

Micromechanical aptasensor-based protein detection using a compact-disc format microfluidics system

A plug-and-play CD-like platform is used to perform a statistical detection of Platelet Derived Growth Factor (PDGF) proteins through aptamer-based surface functionalization of microcantilevers. When PDGF proteins bind to the aptamers, the cantilevers deflect. This deflection is monitored by optical readout heads from a DVD-ROM. The improved sensing platform facilitates measurements in continuous liquid flow with temperature control. Also, the wobbling of the CD platform has been reduced to a minimum and the scanning system has been optimized in order to detect cantilever deflections in liquid in the nanometer range. The capability of the sensing platform is demonstrated by detection of clinically relevant concentrations of PDGF proteins. We have performed statistical measurements on 100 microcantilevers at different concentrations of PDGF, ranging from 10 nM to 400 nM. Hereby it is possible to reliably characterize the averaged mechanical response of cantilevers as a function of protein concentration.

Improving the robustness of Surface Enhanced Raman Spectroscopy based sensors by Bayesian Non-negative Matrix Factorization

Due to applications in areas such as diagnostics and environmental safety, detection of molecules at very low concentrations has attracted recent attention. A powerful tool for this is Surface Enhanced Raman Spectroscopy (SERS) where substrates form localized areas of electromagnetic “hot spots” where the signal-to-noise (SNR) ratio is greatly amplified. However, at low concentrations hot spots with target molecules bound are rare. Furthermore, traditional detection relies on having uncontaminated sensor readings which is unrealistic in a real world detection setting. In this paper, we propose a Bayesian Non-negative Matrix Factorization (NMF) approach to identify locations of target molecules. The proposed method is able to successfully analyze the spectra and extract the target spectrum. A visualization of the loadings of the basis vector is created and the results show a clear SNR enhancement. Compared to traditional data processing, the NMF approach enables a more reproducible and sensitive sensor.