Micromechanical cantilever array sensors for selective fungal immobilization and fast growth detection.
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.
Nugaeva
Natalia
N
Institute of Physics, University of Basel, Klingelbergstrasse 82, Switzerland.
Gfeller
Karin Y
KY
Backmann
Natalija
N
Lang
Hans Peter
HP
Düggelin
Marcel
M
Hegner
Martin
M
D001234:Aspergillus niger / Q000254:growth & development* / Q000302:isolation & purification*; D001422:Bacterial Adhesion / Q000502:physiology; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D015169:Colony Count, Microbial / Q000295:instrumentation* / Q000379:methods; D003199:Computer Systems; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D019563:Mechanics; D008904:Miniaturization; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000254:growth & development* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013172:Spores, Fungal / Q000254:growth & development / Q000302:isolation & purification; D014159:Transducers