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Porous silicon (PSi) has been widely used as a biosensor in recent years due to its large surface area and its optical properties. Most PSi biosensors consist in close-ended porous layers, and, because of the diffusion-limited infiltration of the analyte, they lack sensitivity and speed of response. In order to overcome these shortcomings, PSi membranes (PSiMs) have been fabricated using electrochemical etching and standard microfabrication techniques. In this work, PSiMs have been used for the optical detection of Bacillus cereus lysate. Before detection, the bacteria are selectively lysed by PlyB221, an endolysin encoded by the bacteriophage Deep-Blue targeting B. cereus. The detection relies on the infiltration of bacterial lysate inside the membrane, which induces a shift of the effective optical thickness. The biosensor was able to detect a B. cereus bacterial lysate, with an initial bacteria concentration of 105 colony forming units per mL (CFU/mL), in only 1 h. This proof-of-concept also illustrates the specificity of the lysis before detection. Not only does this detection platform enable the fast detection of bacteria, but the same technique can be extended to other bacteria using selective lysis, as demonstrated by the detection of Staphylococcus epidermidis, selectively lysed by lysostaphin.

A robust fabrication method for stable mesoporous silicon membranes using standard microfabrication techniques is presented. The porous silicon membranes were passivated through the atomic layer deposition of different metal oxides, namely aluminium oxide Al2O3, hafnium oxide HfO2 and titanium oxide TiO2. The fabricated membranes were characterized in terms of morphology, optical properties and chemical properties. Stability tests and optical probing noise level determination were also performed. Preliminary results using an Al2O3 passivated membranes for a biosensing application are also presented for selective optical detection of Bacillus Cereus bacterial lysate. The biosensor was able to detect the bacterial lysate, with an initial bacteria concentration of 106 colony forming units per mL (CFU/mL), in less than 10 min.

Porous silicon (PSi) has been widely used as a biosensor over the last years due to its large surface area and its optical properties. Most PSi biosensors consist in close-ended porous layers, and, because of the diffusion-limited infiltration of the analyte, they lack sensitivity and speed of response. In order to overcome these shortcomings, PSi membranes (PSiMs) have been fabricated using electrochemical etching and standard microfabrication techniques. In this work, PSiMs have been used for the optical detection of Bacillus cereus lysate. Before detection, the bacteria are selectively lysed by PlyB221, an endolysin encoded by the bacteriophage Deep-Blue targeting B. cereus. The detection relies on the infiltration of bacterial lysate inside the membrane, which induces a shift of the effective optical thickness. The biosensor was able to detect a B. cereus bacterial lysate, with an initial bacteria concentration of 106 colony forming units per mL (CFU/mL), in less than 10 min. This work also demonstrates the selectivity of the lysis before detection. Not only does this detection platform enable the fast detection of bacteria, but the same technique can be extended to other bacteria using selective lysis.

In this work, we developed a biosensor for the indirect detection of bacteria via their lysate. The developed sensor is based on porous silicon membranes, which are known for their many attractive optical and physical properties. Unlike traditional porous silicon biosensors, the selectivity of the bioassay presented in this work does not rely on bio-probes attached to the sensor surface; the selectivity is added to the analyte itself, by the addition of lytic enzymes that target only the desired bacteria. The resulting bacterial lysate is then able to penetrate into the porous silicon membrane and affects its optical properties, while intact bacteria accumulate on top of the sensor. The porous silicon sensors, fabricated using standard microfabrication techniques, are coated with TiO 2 layers using atomic layer deposition. These layers serve as passivation but also enhance the optical properties. The performance of the TiO 2 -coated biosensor is tested for the detection of Bacillus cereus, using the bacteriophage-encoded PlyB221 endolysin as the lytic agent. The sensitivity of the biosensor is much improved compared to previous works, reaching 10 3 CFU/mL, with a total assay time of 1 h 30 min. The selectivity and versatility of the detection platform are also demonstrated, as is the detection of B. cereus in a complex analyte.

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: The rapid detection of hazardous bacteria is important for healthcare situations, where such identification can lead to substantial gains for patient treatment and recovery and a reduced usage of broad-spectrum antibiotics. Potential biosensors must be able to provide a fast, sensitive and selective response with as little sample preparation as possible. Indeed, some of these pathogens, such as Staphylococcus aureus, can be yet harmful at very low concentrations in the blood stream, e.g., below 10 colony forming units per mL (CFU/mL). These stringent requirements limit the number of candidates, especially for point-of-care applications. Amongst several biosensing techniques, optical sensing using porous silicon (PSi) substrate has been widely suggested in recent years thanks to unique features such as a large surface area, tunable optical characteristics, and above all relatively easy and affordable fabrication techniques. In most configurations, PSi optical biosensors are close-ended porous layers; this limits their sensitivity and responsiveness due to diffusion-limited infiltration of the analytes in the porous layer. Also, PSi is a reactive material, its oxidation in buffer solutions results in time-varying shifts. Despite its attractive properties, several challenges must still be overcome in order to reach practical applications. Our work addresses three main improvement points. The first one is the stability over time in saline solutions helped by atomic layer deposition of metal oxides inside the pores. Besides a better stability, our solution is helping with an increase of the optical signal to noise ratio, thus reducing the limit of detection. The second one is to perform the lysis of the bacteria prior to its exposure to the sensor, such that the selective detection is based upon the percolation of bacterial residues inside the pores rather than the bacteria themselves. The third one is to remove the bulk silicon below a PSi layer to create a membrane, that allows for flow-through of the analytes, thus enhancing the interactions between the lysate and the sensor’s surface. This approach allows us to avoid the step of surface functionalization used in classical biosensors. We tested thanks to these improvements the selective detection of Bacillus cereus lysate with concentrations between 103 and 105 CFU/mL. Future works are dedicated to further improvements, including optical signal enhancement techniques and dielectrophoretic assisted percolation in the porous silicon membrane.