In-field
analysis (e.g., clinical and diagnostics) using nanostructured
porous silicon (PSi) for label-free optical biosensing has been hindered
so far by insufficient sensitivity of PSi biosensors. Here we report
on a label-free PSi interferometric aptasensor able to specifically
detect tumor necrosis factor alpha (TNFα, a protein biomarker
of inflammation and sepsis) at concentration down to 3.0 nM with signal-to-noise
ratio (S/N) of 10.6 and detection limit (DL) of 200 pM. This represents
a 10 000-fold improvement with respect to direct (i.e., nonamplified)
label-free PSi biosensors and pushes PSi biosensors close to the most
sensitive optical and label-free transduction techniques, e.g., surface
plasmon resonance (SPR) for which a lowest DL of 100 pM in aptasensing
has been reported. A factor 1000 in improvement is achieved by introducing
a novel signal-processing technique for the optical readout of PSi
interferometers, namely, interferogram average over wavelength (IAW)
reflectance spectroscopy. The IAW reflectance spectroscopy is shown
to significantly improve both sensitivity and reliability of PSi biosensors
with respect to commonly used fast Fourier transform (FFT) reflectance
spectroscopy. A further factor 10 is achieved by enabling preparation
of PSi interferometers with enlarged pore sizes (up to a 3× increase
in diameter) at constant current density (i.e., constant porosity
and, in turn, constant refractive index). This method is in contrast
to standard PSi preparation where pore size is increased by increasing
etching current density (i.e., porosity), and allows tackling mass-limited
diffusion of biomolecules into the nanopores without worsening PSi
interferometer optical features.