Low cost easy to use cell viability tests are needed in the pharmaceutical, biomaterial and environmental industry to measure adverse cellular effects. Herein we present a new methodology to track cell death with high resolution. We achieved dynamic digital quantification of cell viability by simple optical imaging using "Single Cell Adhesion Dot Arrays" (SCADA). Fibronectin (FN) dot arrays were fabricated on cell culture multiwell plates. The dot array was designed to accomodate a single cell on each fibronectin dot. For cytotoxicity measurements, cell-filled SCADA substrates were exposed to K2CrO4, HgSO4 salts and dimethyl sulfoxide (DMSO). Adherent cells commonly detach from the surface when they die. Dynamic monitoring of the toxic effect of DMSO and K2CrO4 was done measuring cell detachment rate during more than 30 hours by quantifying the number of occupied dots in the SCADA array. HgSO4 inhibited cellular detachment from the surface, and cytotoxicity was monitored using Trypan Blue life/death assay directly on the surface.In all cases, the cytotoxicity effects were easily monitored with single cell resolution and the results were comparable to previous reports. Cytotoxicity SCADA tests require only a transparent substrate, with a patterned area of less than 1 mm 2 and a reduced number of cells. SCADA enabled dynamic measurements at the highest resolution due to the digital measuring of this methodology. Integrated into microfluidic platforms, SCADA will provide a practical tool that will extent to fundamental research and commercial applications.
The facet of optical fibers coated with nanostructures enable the development of ultraminiature and sensitive (bio)chemical sensors. The reported sensors until now lack of specificity and the fabrication methods offer poor reproducibility. Here, we demonstrate that by transforming the facet of conventional multimode optical fibers onto plasmon resonance energy transfer (PRET) antenna surfaces the specificity issues may be overcome. To do so, a low cost chemical approach was developed to immobilize gold nanoparticles on the optical fiber facet in a reproducible and controlled manner. Our nanosensors are highly selective as PRET is a nanospectroscopic effect that only occurs when the resonant wavelength of the nanoparticles matches that of the target parameter. As an example, we demonstrate the selective detection of picomolar concentrations of copper ions in water. Our sensor is 1,000 times more sensitive than state of the art technologies. An additional advantage of our nanosensors is their simple interrogation; it comprises of a lowpower light emitting diode, a multimode optical fiber coupler, and a miniature spectrometer. We believe that the PRET-based fiber optic platform reported here may pave the way of the development of a new generation of ultra-miniature, portable, and hypersensitive and selective (bio)chemical sensors.
Optical fiber-based Localized Surface Plasmon Resonance (OF-LSPR) biosensors have emerged as an ultra-sensitive miniaturized tool for a great variety of applications. Their fabrication by the chemical immobilization of gold nanoparticles (AuNPs) on the optic fiber end face is a simple and versatile method. However, it can render poor reproducibility given the number of parameters that influence the binding of the AuNPs. In order to develop a method to obtain OF-LSPR sensors with high reproducibility, we studied the effect that factors such as temperature, AuNPs concentration, fiber core size and time of immersion had on the number and aggregation of AuNPs on the surface of the fibers and their resonance signal. Our method consisted in controlling the deposition of a determined AuNPs density on the tip of the fiber by measuring its LSPR signal (or plasmonic signal, Sp) in real-time. Sensors created thus were used to measure changes in the refractive index of their surroundings and the results showed that, as the number of AuNPs on the probes increased, the changes in the Sp maximum values were ever lower but the wavelength shifts were higher. These results highlighted the relevance of controlling the relationship between the sensor composition and its performance.
A sensor based on gold nanoparticles immobilized onto an optical fiber facet is shown. The innovation is related to the plasmon resonance energy transfer phenomenon application, demonstrating its usefulness for Cytochrome C detection.
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