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Orthogonal subspace signal processing algorithms (OSPA) have been developed to extract the optical thickness of a porous silicon layer to within one part in 10 5 from its reflectivity spectrum. This is equivalent to a limit of detection (LOD) of -40 pm change in optical thickness for a 3 μm thick layer, or an LOD of 1/2000 of a monolayer coverage with antibodies, of molecular weight 160 k Daltons, within a layer with pores of 100 nm diameter. A large molecule {horseradish peroxidase (HRP), MWt 40 kDa} has been detected at a concentration of 1 μg/ml by measuring its direct binding to anti-HRP antibodies immobilised within a porous silicon layer. A competitive assay has been demonstrated for the detection of a small molecule {2, 4, 6 trinitrotoluene (TNT), MWt 227 Da} at 10 μg/ml. The projected LODs for HRP and TNT by these assays are 50 ng/ml and 1 μg/ml respectively.

The sensitivity of the optical reflectivity of porous silicon structures to the refractive index of liquid within the pores is studied for a single layer, a Bragg mirror and a microcavity. Sucrose solutions of concentration in the range 0.05 to 1.0% by weight are introduced into the pores within a flow cell in order to change the refractive index of the liquid in the pores from 1.3330 to 1.3344. Optimum wavelengths for detection via reflectivity changes are determined based on a signal to noise analysis. The optical thickness of the single layer is also monitored by measuring the fringe spacing via a Fourier transform technique. It is just possible to detect the effect of a change in refractive index of liquid in the pores of 0.00007 for both the reflectivity and optical thickness approaches.

To use porous silicon as an optical interferometric biosensor, the pores must be sufficiently large to allow easy ingress of reagents and the layer must also display Fabry-Perot optical cavity modes. Here the detection antibody is rabbit IgG and the analyte is α-rabbit IgG conjugated to horseradish peroxidase (HRP). For this model system, the pores should be >50 nm in diameter. Such diameters have been obtained in 0.05 Ω cm n-type silicon using anodisation followed by chemical etching in ethanolic KOH and also by anodising 0.005 Ω cm p-type material. The latter also displays optical cavity modes. The silicon surface is oxidised in ozone, silanised using aminopropylmethoxysilanes with one, two or three methoxy groups, and cross linked to IgG using glutaraldehyde. High specific binding is found for mono-, di- and tri-methoxy silanes, but the lowest non-specific binding is found for silanisation with the tri-methoxy silane.