Fluorescence Detection of DNA Hybridization Using an Integrated Thin-Film Amorphous Silicon n-i-p Photodiode

ABSTRACTThis paper presents the fluorescence detection of DNA hybridization with a surface immobilized probe using an hydrogenated amorphous silicon photosensor. This sensor integrates a SiO2 layer for DNA probe immobilization, a p-i-n amorphous silicon (a-Si:H) photodiode for fluorescence detection and a fluorescence filter of amorphous silicon carbon (a-SiC:H) to cut the excitation light. With this integrated photosensor system, a five order of magnitude difference was obtained in the signal measured at the emission wavelength and that measured at the excitation wavelength for the same incident photon flux. The fluorophore Alexa Fluor 430 was used to label the DNA target molecules and a laser at 405 nm and a photon flux of 5.7×1016 cm−2.s−1 was used as the excitation light source. The detection limit achieved for fluorophores in solution in contact with the device and for fluorophores immobilized on the device surface is 5×10−9 M and 0.4 pmol/cm2, respectively. The fluorescence detection of the DNA target hybridization with a covalently or electrostatically immobilized probe was successfully detected at a surface density of ∼3 pmol/cm2.