Micromechanical cantilever array sensors for selective fungal immobilization and fast growth detection

• DOI: 10.1016/j.bios.2005.02.004
• Publication Date: 2005-03-03T13:14:28Z
• Authors: Natalia Nugaeva; Karin Y. Gfeller; Natalija Backmann; Hans Peter Lang; Marcel Düggelin; Martin Hegner
• Keywords: Miniaturization; Transducers; Cell Culture Techniques; Colony Count, Microbial; Reproducibility of Results; Biosensing Techniques; Equipment Design; Saccharomyces cerevisiae; 02 engineering and technology; Spores, Fungal; Mechanics; Sensitivity and Specificity; Bacterial Adhesion; Equipment Failure Analysis; Computer Systems; Electrochemistry; Aspergillus niger; 0210 nano-technology; Cell Proliferation

We demonstrate the use of micromechanical cantilever arrays for selective immobilization and fast quantitative detection of vital fungal spores. Micro-fabricated uncoated as well as gold-coated silicon cantilevers were functionalized with concanavalin A, fibronectin or immunoglobulin G. In our experiments two major morphological fungal forms were used--the mycelial form Aspergillus niger and the unicellular yeast form Saccharomyces cerevisiae, as models to explore a new method for growth detection of eukaryotic organisms using cantilever arrays. We exploited the specific biomolecular interactions of surface grafted proteins with the molecular structures on the fungal cell surface. It was found that these proteins have different affinities and efficiencies to bind the spores. Maximum spore immobilization, germination and mycelium growth was observed on the immunoglobulin G functionalized cantilever surfaces. We show that spore immobilization and germination of the mycelial fungus A. niger and yeast S. cerevisiae led to shifts in resonance frequency within a few hours as measured by dynamically operated cantilever arrays, whereas conventional techniques would require several days. The biosensor could detect the target fungi in a range of 10(3) - 10(6) CFUml(-1). The measured shift is proportional to the mass of single fungal spores and can be used to evaluate spore contamination levels. Applications lie in the field of medical and agricultural diagnostics, food- and water-quality monitoring.