Detection of biological macromolecules on a biochip dedicated to UV specific absorption.
This work describes an ultraviolet biosensing technique based on specific molecular absorption detected with a previously developed spectrally selective aluminum gallium nitride (AlGaN) based detector. Light absorption signal of DNA and proteins, respectively at 260 nm and 280 nm, is used to image biochips. To allow detection of protein or DNA monolayers at the surface of a biochip, we develop contrast-enhancing multilayer substrates. We analyze them through models and experiments and validate the possibility of measuring absorptions of the order of 10(-3). These multilayer structures display a high reflectivity, and maximize the interaction of the electric field with the biological element at the chip surface. Optimization of the experimental absorption, which includes effects such as roughness of the biochip, spectral and angular resolution of the optics, illumination, etc., is carried out with an inorganic ultraviolet absorber (titanium dioxide) deposit. We obtained an induced absorption contrast enhanced by a factor of 4.0, conferring enough sensitivity to detect monolayers of DNA or proteins. Experimental results on an Escherichia coli histidine-tagged methionyl-tRNA synthetase protein before and after complexation with an anti-polyHis specific antibody validate our biosensing technique. This label-free optical method may be helpful in controlling biochip coatings, and subsequent biological coupling at the surface of a biochip.
Robin
Kristelle
K
Thales Research and Technology, Palaiseau Cedex, France. kristelle.robin@thalesgroup.com
Reverchon
Jean-Luc
JL
Mugherli
Laurent
L
Fromant
Michel
M
Plateau
Pierre
P
Benisty
Henri
H
D015374:Biosensing Techniques / Q000295:instrumentation*; D004247:DNA / Q000032:analysis*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D056656:Lab-On-A-Chip Devices; D046911:Macromolecular Substances / Q000032:analysis; D010783:Photometry / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation* / Q000379:methods
D046911:Macromolecular Substances; D011506:Proteins; D004247:DNA