Abstract:We present experimental results showing that long-period gratings in photonic crystal fibers can be used as sensitive biochemical sensors. A layer of biomolecules was immobilized on the sides of the holes of the photonic crystal fiber and by observing the shift in the resonant wavelength of a long-period grating it was possible to measure the thickness of the layer. The long-period gratings were inscribed in a large-mode area silica photonic crystal fiber with a CO 2 laser. The thicknesses of a monolayer of poly-L-lysine and double-stranded DNA was measured using the device. We find that the grating has a sensitivity of approximately 1.4nm/1nm in terms of the shift in resonance wavelength in nm per nm thickness of biomolecule layer.

Abstract:We demonstrate evanescent-wave sensing of Cy5-DNA-molecules in an aqueous solution using a photonic crystal fiber. Less than 0.8pL sample volume placed in the holes of the fiber is sufficient for reliable detection.

We demonstrate selective detection of fluorophore labeled antibodies from minute samples probed by a sensor layer of complementary biomolecules immobilized inside the air holes of microstructured Polymer Optical Fiber (mPOF). The fiber core is defined by a ring of 6 air holes and a simple procedure was applied to selectively capture either alpha-streptavidin or alpha-CRP antibodies inside these air holes. A sensitive and easy-to-use fluorescence method was used for the optical detection. Our results show that mPOF based biosensors can provide reliable and selective antibody detection in ultra small sample volumes.

It was previously shown that enhanced nisin resistance in some mutants was associated with increased expression of three genes, pbp2229, hpk1021, and lmo2487, encoding a penicillin-binding protein, a histidine kinase, and a protein of unknown function, respectively. In the present work, we determined the direct role of the three genes in nisin resistance. Interruption of pbp2229 and hpk1021 eliminated the nisin resistance phenotype. Interruption of hpk1021 additionally abolished the increase in pbp2229 expression. The results indicate that this nisin resistance mechanism is caused directly by the increase in pbp2229 expression, which in turn is brought about by the increase in hpk1021 expression. We also found a degree of cross-protection between nisin and class IIa bacteriocins and investigated possible mechanisms. The expression of virulence genes in one nisin-resistant mutant and two class IIa bacteriocin-resistant mutants of the same wild-type strain was analyzed, and each mutant consistently showed either an increase or a decrease in the expression of virulence genes (prfA-regulated as well as prfA-independent genes). Although the changes mostly were moderate, the consistency indicates that a mutant-specific change in virulence may occur concomitantly with bacteriocin resistance development.Nisin and the class IIa bacteriocins (also called pediocin-like bacteriocins) are antimicrobial peptides that are produced by lactic acid bacteria and that have the greatest potential as biopreservatives for food. One of the main target organisms in this context is Listeria monocytogenes, a food-borne pathogen that causes severe human illness as well as economic losses for the food industry.Nisin exerts its antimicrobial action by forming pores in the cytoplasmic membrane through an interaction with the peptidoglycan precursor lipid II (for a recent review, see reference 17). Enhanced nisin resistance in L. monocytogenes generally constitutes less than a 10-fold increase in the MIC. Nisin resistance in several, but not all, spontaneous mutants of L.

In this paper we present the first incorporation of a microstructured optical fiber (MOF) into biochip applications. A 16-mm-long piece of MOF is incorporated into an optic-fluidic coupler chip, which is fabricated in PMMA polymer using a CO(2) laser. The developed chip configuration allows the continuous control of liquid flow through the MOF and simultaneous optical characterization. While integrated in the chip, the MOF is functionalized towards the capture of a specific single-stranded DNA string by immobilizing a sensing layer on the microstructured internal surfaces of the fiber. The sensing layer contains the DNA string complementary to the target DNA sequence and thus operates through the highly selective DNA hybridization process. Optical detection of the captured DNA was carried out using the evanescent-wave-sensing principle. Owing to the small size of the chip, the presented technique allows for analysis of sample volumes down to 300 nL and the fabrication of miniaturized portable devices.