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Biopolymer-based packaging films were prepared from pectin (PEC) and sodium alginate (SA), with the incorporation of 10 % MCC and different concentrations of geraniol (GER at 2.5, 5.0, 7.5, and 10.0 %). Rheological properties suggested that film-forming solutions and film-forming emulsions exhibited a shear-thinning or pseudo-plastic non-Newtonian behaviour. The dried films were crosslinked with 2.0 % CaCl 2 . The addition of MCC into PEC/SA film enhanced the TS but reduced it with the impregnation of GER without influencing the EAB and toughness of the film. The water solubility of the films significantly reduced with the rise in the GER levels but enhanced the water vapor and oxygen barrier attributes. TGA demonstrated that incorporating MCC reduced the film's thermal degradation (44.92 % to 28.81 %), but GER had an insignificant influence on the thermal stability. FTIR spectra revealed that hydrogen bond formation was positively linked with the GER addition in the film formulation. X-ray diffractograms showed that prepared films were predominantly amorphous. Antimicrobial studies showed a complete reduction of Escherichia coli and Bacillus cereus in 24 h. Overall, the composite film displayed excellent physical and active properties and PEC/SA/MCC/5.0 %GER/CaCl 2 film was considered the best formulation for food packaging applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The present work investigated the structure-function relationship of dry fractionated oat flour (DFOF) as a techno-functional ingredient using bread as a model system. Mechanically, DFOF fractions (F), that is, F1: <224 µm, F2: 250-280 µm, F3: 280-500 µm, F4: 500-600 µm, and whole oat flour (F5) were blended with white wheat flour at 10%, 30%, and 50% substitution levels for bread making. The blended flours, doughs, and bread samples were assessed for their techno-functional, nutritional, and structural characteristics. The results of Mixolab and the Rapid Visco Analyzer show that the 50% substituted F3 fraction exhibits the highest water absorption properties (69.53%), whereas the 50% F1 fraction exhibits the highest peak viscosity of the past slurry. Analysis of bread samples revealed a lower particle size of DFOF fractions and higher supplementation levels, increased β-glucan levels (0.13-1.29 g/100 bread (db), reduced fermentable monosaccharides, that is, glucose (1.44-0.33 g/100 g), and fructose (1.06-0.28 g/100 g). The effect of particle size surpassed the substitution level effect on bread volume reduction. The lowest hardness value for F1 is 10%, and the highest value for F2 is 50%. The total number of cells in the bread slice decreased from the control to the F4 fraction (50%). Multi-criteria analysis indicated that DFOF fractions produced breads with similar structure and higher nutritional value developed from white wheat flour. PRACTICAL APPLICATION: The use of mechanically fractionated oat flours fractions in white wheat flour breads can improve the nutritional profile without affecting the physical properties of the bread product. Based on the oat flour fractions, bakers and food processing companies can tailor the bread formulations for high β-glucan, high fiber, and low reduced sugar claims., (© 2024 The Authors. Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

The market has observed a rapid increase in the demand for plant-based foods as an alternative to animal meat products. Technologies such as high-moisture extrusion (HME) have the potential to develop anisotropic structures using alternative protein ingredients. This article discusses the different possible mechanisms responsible for structure formation and the effect of extrusion process parameters and outlines the recent advances in the long cooling dies (LCDs) used for meat alternative development. The role of different protein ingredients and the impact of combining them with other biopolymers were also evaluated. The underlying mechanism behind anisotropic structure formation during HME is a synergistic effect, with substantial dependence on the source of ingredients and their processing background. Formulation including proteins derived from plants, insects, animals, and microalgae with other biopolymers could pave the way to develop structured meat alternatives and fill nutritional interstices. Dynamic or rotating annular gap cooling dies operating at freely controllable shear and static annular gap dies are recent developments and assist to produce layered or fibrous structures. The complex chemical sites created during the HME of plant protein favour flavour and colour retention. This paper summarises the recent information published in the scientific literature and patents, which could further help researchers to fill the present knowledge gaps.

Suitable packaging material in combination with high-pressure processing (HPP) can retain nutritional and organoleptic qualities besides extending the product's shelf life of food products. However, the selection of appropriate packaging materials suitable for HPP is tremendously important because harsh environments like high pressure and high temperature during the processing can result in deviation in the visual and functional properties of the packaging materials. Traditionally, fossil-based plastic packaging is preferred for the HPP of food products, but these materials are of serious concern to the environment. Therefore, bio-based packaging systems are proposed to be a promising alternative to fossil-based plastic packaging. Some studies have scrutinized the impact of HPP on the functional properties of biopolymer-based packaging materials. This review summarizes the HPP application on biopolymer-based film-forming solutions and pre-formed biopolymer-based films. The impact of HPP on the key packaging properties such as structural, mechanical, thermal, and barrier properties in addition to the migration of additives from the packaging material into food products were systemically analyzed. HPP can be applied either to the film-forming solution or preformed packages. Structural, mechanical, hydrophobic, barrier, and thermal characteristics of the films are enhanced when the film-forming solution is exposed to HPP overcoming the shortcomings of the native biopolymers-based film. Also, biopolymer-based packaging mostly PLA based when exposed to HPP at low temperature showed no significant deviation in packaging properties indicating the suitability of their applications. HPP may induce the migration of packaging additives and thus should be thoroughly studied. Overall, HPP can be one way to enhance the properties of biopolymer-based films and can also be used for packaging food materials intended for HPP.

Background: Many adolescents meeting diagnostic criteria for social anxiety disorder (SAD) do not seek help or access timely professional supports. The current study investigated two hypothesized barriers to adolescents' seeking help for SAD: stigma and SAD symptoms., Method: Three hundred fifteen adolescents (52% female; mean age = 14.85 years) read a vignette describing a hypothetical peer with symptoms of SAD and completed measures of their own help-seeking intentions, SAD symptoms, stigma awareness and agreement., Results: The majority of adolescents reported that the vignette character needed help. Adolescents with more SAD symptoms, and those with greater stigma awareness, were less likely to indicate they would seek help from an adult they know (e.g., parent or teacher) for similar symptoms., Conclusions: Both SAD symptoms and perceptions of societal stigma may act as barriers to help-seeking for young people indicating the importance of intervention with young people, and the adults with whom they interact regularly., (© 2021 The Authors. Early Intervention in Psychiatry published by John Wiley & Sons Australia, Ltd.)

Background: Pregestational diabetes mellitus (PGDM) is associated with adverse pregnancy outcomes. Studies assessing interventions to improve maternal and infant outcomes have increased exponentially over recent years. Several outcomes in this field of maternal diabetes are rare, making it difficult to synthesise evidence., Objectives: To collect outcomes reported in studies assessing treatment interventions in pregnant women with PGDM., Search Strategy: CENTRAL, Web of Science, Medline, CINAHL, Embase and ClinicalTrials.gov from their inception until 27 January 2020., Selection Criteria: Any randomised controlled trial assessing treatment interventions in pregnant women with PGDM reported in English., Data Collection and Analysis: Two independent reviewers assessed the suitability of articles and retrieved the data. Outcomes extracted from the literature were broadly categorised into maternal, fetal/infant or other outcomes by the study advisory group., Main Results: Sixty-seven of the 1475 studies identified fulfilled the inclusion criteria. The median number of outcomes reported per study was 15 (range 1-46). The majority of studies were from North America and Europe. Insulin and metformin were the most commonly investigated pharmacological interventions. Glucose monitoring was the most assessed technological intervention. In all, 131 unique outcomes were extracted: maternal (n = 69), fetal/infant (n = 61) and other (n = 1)., Conclusions: Outcome reporting in treatment interventions trials of pregnant women with PGDM is varied, making it difficult to synthesise evidence, especially for rare outcomes. Systems are needed to standardise outcome reporting in future clinical trials and so facilitate evidence synthesis in this area of maternal diabetes., Registration: The systematic review was registered prospectively with the International Prospective Register of Systematic Reviews (PROSPERO) database (Registration number CRD42020173549)., Tweetable Abstract: Outcome reporting is heterogeneous in intervention trials of pregnant women with diabetes existing before pregnancy., (© 2021 The Authors. BJOG: An International Journal of Obstetrics and Gynaecology published by John Wiley & Sons Ltd.)

Objective: To develop a core outcome set (COS) for randomised controlled trials (RCTs) evaluating the effectiveness of interventions for the treatment of pregnant women with pregestational diabetes mellitus (PGDM)., Design: A consensus developmental study., Setting: International., Population: Two hundred and five stakeholders completed the first round., Methods: The study consisted of three components. (1) A systematic review of the literature to produce a list of outcomes reported in RCTs assessing the effectiveness of interventions for the treatment of pregnant women with PGDM. (2) A three-round, online eDelphi survey to prioritise these outcomes by international stakeholders (including healthcare professionals, researchers and women with PGDM). (3) A consensus meeting where stakeholders from each group decided on the final COS., Main Outcome Measures: All outcomes were extracted from the literature., Results: We extracted 131 unique outcomes from 67 records meeting the full inclusion criteria. Of the 205 stakeholders who completed the first round, 174/205 (85%) and 165/174 (95%) completed rounds 2 and 3, respectively. Participants at the subsequent consensus meeting chose 19 outcomes for inclusion into the COS: trimester-specific haemoglobin A1c, maternal weight gain during pregnancy, severe maternal hypoglycaemia, diabetic ketoacidosis, miscarriage, pregnancy-induced hypertension, pre-eclampsia, maternal death, birthweight, large for gestational age, small for gestational age, gestational age at birth, preterm birth, mode of birth, shoulder dystocia, neonatal hypoglycaemia, congenital malformations, stillbirth and neonatal death., Conclusions: This COS will enable better comparison between RCTs to produce robust evidence synthesis, improve trial reporting and optimise research efficiency in studies assessing treatment of pregnant women with PGDM., Tweetable Abstract: 165 key stakeholders have developed #Treatment #CoreOutcomes in pregnant women with #diabetes existing before pregnancy., (© 2021 The Authors. BJOG: An International Journal of Obstetrics and Gynaecology published by John Wiley & Sons Ltd.)

Background: The stigma associated with mental disorders in adolescence has a range of detrimental consequences, negatively impacting help-seeking behaviours and quality of life. Social anxiety typically has its onset during adolescence, but the associated stigma is not well understood. This study was designed to improve understanding of social anxiety and depression stigma in adolescence by examining demographic and personal variables that predict them both., Methods: Three hundred and fifteen adolescents (150 males, 165 females), from 2nd and 4th year of secondary school (mean ages 13.92 and 15.92 years respectively), completed the Peer Mental Health Stigmatization Scale to measure both personal and perceived stigma towards vignettes depicting social anxiety and depression. They also answered demographic questions and completed the Social Anxiety Scale for Adolescents., Results: Hierarchical multiple regression revealed that gender predicted both personal and perceived stigma towards depression and personal stigma towards social anxiety. Males scored higher on measures of personal stigma whereas females had higher scores on perceived stigma. Ability to accurately identify symptoms described in a vignette, predicted lower personal stigma towards social anxiety and higher perceived stigma towards depression. Depression was more stigmatized than social anxiety., Limitations: The study used vignettes to represent individuals with depression and social anxiety so participant responses may not reflect their behaviour towards real peers., Conclusions: Findings emphasise the importance of separately considering personal and perceived stigma for each condition and highlight gender differences in stigma responses that need to be considered in intervention design., (Copyright © 2020. Published by Elsevier B.V.)

A 73-year-old female patient with epilepsy presented to hospital with a progressive, diffuse macular rash over the trunk and limbs with associated mucosal blistering and discharge. Ocular symptoms initially predominated and she was treated for presumed bacterial conjunctivitis by her General Practitioner the previous day. On the acute medical unit supportive management was initiated for suspected adverse drug reaction (ADR) to a recent lamotrigine dose increase. Skin biopsy confirmed a diagnosis of toxic epidermal necrolysis. We present this case to highlight the importance of medication history taking and raise awareness of indolent presentations of life-threatening ADRs. Caution should be applied following dose changes to anti-epileptics, even if previously stable.

A 73-year-old female attended the Emergency Department with a twenty four hour history of a progressive, diffuse macular rash, predominantly affecting limbs and trunk, with associated oral and ocular discharge.

Core Outcome Sets (COS) contain an agreed minimum set of outcomes to be measured and reported in all studies in a specific area, with the objective of standardizing outcome reporting. COS may minimize research waste by identifying outcomes important to key stakeholders, allowing for improved evidence synthesis, and facilitating translation of research findings to clinical practice. Over the past 5 years, there has been significant progress in developing COS relevant to studies of diabetes in pregnancy. This review summarizes work in this area, reviews the role of patient and public involvement in COS development, and suggests areas for future research., (© 2020 by the American Diabetes Association.)

Materials have been developed that encapsulate a homogeneous catalyst and enable it to operate as a heterogeneous catalyst in water. A hydrophobic ionic liquid within the material was used to dissolve Fe-TAML and keep it from leaching into the aqueous phase. One-pot processes were used to entrap Fe-TAML in basic ionic liquid gels, and ionic liquid gel spheres structured via a modified Stöber synthesis forming SiO 2 particles of uniform size. Catalytic activity was demonstrated via the oxidative degradation of dyes. Fe-TAML entrapped in a basic ionic liquid gel exhibited consistent activity in five recycles. This discovery of heterogenized H 2 O 2 activators prepared by sol-gel and Stöber processes opens new possibilities for the creation of engineered catalytic materials for water purification.

This article reports on a fiber-based ratiometric optical pH sensor for use in real-time and continuous in vivo pH monitoring in human tissue. Stable hybrid sol-gel-based pH sensing material is deposited on a highly flexible plastic optical fiber tip and integrated with excitation and detection electronics. The sensor is extensively tested in a laboratory environment before it is applied in vivo in a human model. The pH sensor performance in the laboratory environment outperforms the state-of-the-art reported in the current literature. It exhibits the highest sensitivity in the physiological pH range, resolution of 0.0013 pH units, excellent sensor to sensor reproducibility, long-term stability, short response time of <2 min, and drift of 0.003 pH units per 22 h. The sensor also exhibits promising performance in in vitro whole blood samples. In addition, human evaluations conducted under this project demonstrate successful short-term deployment of this sensor in vivo., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Plasmonic cavities are widely studied for the large amplifications in fluorophore emission intensity that they can achieve. Exploiting these properties for biological sensing applications requires strategies to selectively insert the target antigen into a resonant cavity, which are often of similar size or smaller than the target molecule. Here we demonstrate that using relatively simple solution processing, cavity structures can be grown at the stochastic locations where antigen binding takes place, which yields large enhancements in fluorophore emission intensities and over an order of magnitude improvement in bioassay response. The fluorescence amplification generated by the in situ growth of plasmonic structures is sufficiently large to enable both single antigen and single low-quantum efficiency fluorophore detection. The simplicity of the process demonstrated negates the requirement for complex surfaces or geometries and is readily adaptable for a wide range of diagnostic applications.

Rapid tranquillization is a pharmacological intervention sometimes employed in mental health care for the management of acute behavioural disturbance. It is a form of restrictive practice, which, along with seclusion and restraint, is a conventional and controversial intervention in the therapeutic management of risk in mental health settings. This study surveyed mental health nurses practice in rapid tranquillization. A self-report questionnaire was utilized which addressed aspects such as definitions of rapid tranquillization, presence of rapid tranquillization policy, types of incidents where it is used and postintervention monitoring. The results demonstrate that rapid tranquillization is an intervention used in the management of acute behavioural disturbance in various mental health settings in Ireland. Respondents showed a basic understanding of rapid tranquillization as an intervention; however, some areas reported not having a specific rapid tranquillization policy. There was some evidence of a variation in postrapid tranquillization monitoring of psychiatric/mental health and physical health. Service user debriefing following rapid tranquillization was reported to be common; however, the content of this was not elaborated on. In the light of variations in practice, specific training and the development of rapid tranquillization policies are recommended., (© 2018 Australian College of Mental Health Nurses Inc.)

In this work, ultra-bright fluorescent silica nanoparticles (NPs) labels have been shown to selectively bind to a model circulating tumour cell (CTC) line, MCF-7, a metastatic breast cancer by targeting epithelial cellular adhesion molecule (EpCAM) present on the MCF-7 cell membrane. Silica NPs approximately 40nm in diameter were doped with different concentrations of Cyanine5 dye molecules, using the reverse microemulsion method. The NPs were two orders of magnitude brighter than Cyanine5 free dye and the measured fluorescence intensity matched a homo-Förster Resonance Energy Transfer model. NPs were conjugated with anti-EpCAM antibody to the NP surface for immunospecific targeting. In flow cytometry experiments the NPs were twice as bright as two commercial anti-EpCAM red fluorophore conjugates, APC and AlexaFluor®647. This increase is achieved while keeping non-specific binding low as established in control tests with a non-metastatic cancer cell line (HeLa). The NPs were also immunospecific in fluorescence microscopy experiments performed at room temperature on non-fixed cells. Confocal microscopy was used to confirm the NPs were located on the surface of the cells, matching with the location of the EpCAM marker. These NPs labels have excellent potential in biomedical diagnostics, particularly when high signal to noise and good photostability are needed, for example, in point-of-care testing., (Copyright © 2016 Elsevier B.V. All rights reserved.)

We demonstrate a method to maximise the fluorescence enhancement from a dye using gold coated diffraction gratings. Rotations about the azimuth provides a convenient approach to maximise the coupling between the grating and excitation source while achieving enhancements comparable to traditional optical configurations where the grating and in plane light vectors are parallel. This approach yields a 30 fold enhancement in the fluorescence signal over metal free substrates, while opening up the range of possible orientations and configurations suitable for fluorescence enhancement applications.

Herein we report the use of high brightness Cyanine5-doped silica nanoparticles (NPs) for the detection of antibodies or DNA in microarray bioassays. NP labels showed negligible non-specific binding, greater sensitivity and lower limits of detection when compared to free dye-labelled biomolecules. Moreover, the spotted microarrays used in this study required low NP and antibody concentrations to generate large data sets with improved statistical accuracy. These NPs have significant potential for use in biosensing for disease detection.

We report a new flow control method for centrifugal microfluidic systems; CO₂ is released from on-board stored baking powder upon contact with an ancillary liquid. The elevated pressure generated drives the sample into a dead-end pneumatic chamber sealed by a dissolvable film (DF). This liquid incursion wets and dissolves the DF, thus opening the valve. The activation pressure of the DF valve can be tuned by the geometry of the channel upstream of the DF membrane. Through pneumatic coupling with properly dimensioned disc architecture, we established serial cascading of valves, even at a constant spin rate. Similarly, we demonstrate sequential actuation of valves by dividing the disc into a number of distinct pneumatic chambers (separated by DF membranes). Opening these DFs, typically through arrival of a liquid to that location on a disc, permits pressurization of these chambers. This barrier-based scheme provides robust and strictly ordered valve actuation, which is demonstrated by the automation of a multi-step/multi-reagent DNA-based hybridization assay.

Fluorescent silica nanoparticle (NP) labels are of great interest in biomedical diagnostics, however, when used in bioassays under physiological conditions they rapidly agglomerate and precipitate from solution leading to high levels of non-specific binding. In this work, using size and zeta-potential data obtained from Dynamic and Electrophoretic Light Scattering analysis, the improvement in colloidal stability of silica NPs under physiological conditions was correlated with an increase in the concentration of three additives: (1) a protein, bovine serum albumin (BSA); (2) a neutral surfactant, Tween 20®; and (3) a charged surfactant, sodium dodecyl sulfate (SDS). The number of BSA molecules present in the NP corona at each concentration was calculated using UV-Vis spectroscopy and a bicinchoninic acid protein assay (BCA). The optimal concentration of each additive was also effective in stabilizing antibody labeled fluorescent nanoparticles (αNPs) under physiological conditions. Using a fourth additive, trehalose, the colloidal stability of αNPs after freeze-drying and long-term storage also significantly improved. Both as-prepared and freeze-dried αNPs were tested in a standard fluorescence immunoassay for the detection of human IgG. The as-prepared assay showed a higher sensitivity at low concentration and a lower limit of detection when compared to a free dye assay. Assays performed with freeze dried αNPs after 4 and 22 days also showed good reproducibility., (Copyright © 2015 Elsevier Inc. All rights reserved.)

This paper describes the fabrication of oligonucleotide-coated Cy5-doped silica nanoparticles using a combination of multivalent linkers and their use in surface-based DNA sandwich hybridization assays. Dipodal silane is introduced as a means to fabricate amine-coated silica nanoparticles and its advantages compared to monopodal silanes are discussed. The use of dipodal silane in conjunction with three different polymer linkers (oxidized dextran, linear and 8-arm polyethylene glycol (PEG)) to immobilize single-stranded DNA to Cy5-doped nanoparticles is investigated and dynamic light scattering measurements and Fourier transform infrared spectroscopy are used to follow the progression of the functionalization of the nanoparticles. We observe a significant improvement in the binding stability of the single-stranded DNA when the dipodal silane and 8-arm PEG are used in combination, when compared to alternative conjugation strategies. Both 8mer and 22mer oligonucleotides are securely conjugated to the high-brightness nanoparticles and their availability to hybridize with a complementary strand is confirmed using solution-based DNA hybridization experiments. In addition, a full surface-based sandwich assay demonstrates the potential these nanoparticles have in the detection of less than 500 femtomolar of a DNA analogue of micro RNA, miR-451.

Human cancers are incredibly diverse with regard to molecular aberrations, dependence on oncogenic signaling pathways, and responses to pharmacological intervention. We wished to assess how cellular dependence on the canonical PI3K vs. MAPK pathways within HER2+ cancers affects responses to combinations of targeted therapies, and biomarkers predictive of their activity. Through an integrative analysis of mechanistic model simulations and in vitro cell line profiling, we designed a six-arm decision tree to stratify treatment of HER2+ cancers using combinations of targeted agents. Activating mutations in the PI3K and MAPK pathways (PIK3CA and KRAS), and expression of the HER3 ligand heregulin determined sensitivity to combinations of inhibitors against HER2 (lapatinib), HER3 (MM-111), AKT (MK-2206), and MEK (GSK-1120212; trametinib), in addition to the standard of care trastuzumab (Herceptin). The strategy used to identify effective combinations and predictive biomarkers in HER2-expressing tumors may be more broadly extendable to other human cancers.

In this work we present a centrifugal microfluidic system enabling highly efficient collective trapping and alignment of particles such as microbeads and cells, their multi-colour fluorescent detection and subsequent manipulation by optical tweezers. We demonstrate array-based capture and imaging followed by "cherry-picking" of individual particles, first for fluorescently labelled polystyrene (PS) beads and then for cells. Different cell lines are discriminated based on intracellular as well as surface-based markers.

Carpal tunnel syndrome (CTS) is the most common entrapment neuropathy, affecting 9% of women, and it is responsible for significant morbidity and occupational absence. Clinical assessment is used for initial diagnosis and nerve conduction (NC) studies are currently the principal test used to confirm the diagnosis. Sensitivity of NC studies is >85% and specificity is >95%. There is now good evidence that US can be used as an alternative to NC studies to diagnose CTS. US can assess the anatomy of the median nerve and also identify pathology of the surrounding structures that may compress the nerve. Median nerve enlargement (cross-sectional area ≥10 mm(2) at the level of the pisiform bone or tunnel inlet) is the most commonly used parameter to diagnose CTS on US, and sensitivity has been reported to be as high as 97.9% using this parameter. US may also be used to guide therapeutic corticosteroid injection into the carpal tunnel--thus avoiding median nerve injury--and to objectively monitor the response to treatment. There is now sufficient evidence to propose a new paradigm for the diagnosis of CTS that incorporates US. US is proposed as the initial diagnostic test in CTS based on similar sensitivity and specificity to NC studies but higher patient acceptability, lower cost and additional capability to assess carpal tunnel anatomy and guide injection., (© The Author 2014. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

An in-depth understanding of biochemical processes occurring within biological systems is key for early diagnosis of disease and identification of appropriate treatments. Nanobiophotonics offers huge potential benefits for intracellular diagnostics and therapeutics. Intracellular sensing using fluorescent nanoparticles is a potentially useful tool for real-time, in vivo monitoring of important cellular analytes. This work is focused on synthesis of optical chemical nanosensors for the quantitative analysis of pH inside living cells. The structure of the nanosensor comprises a biofriendly silica matrix with co-encapsulated Texas Red, acting as a reference dye, and pH-sensitive fluorescein isothiocyanate enabling ratiometric quantitative environmental detection. In order to obtain silica-based nanoparticles -70 nm in size, a modified sol-gel-based Stöber method was employed. The potential of these nanosensors for intracellular pH monitoring is demonstrated inside a live human embryonic kidney cell line whereby a significant change in fluorescence is observed when the cell pH is switched from acidic to basic. High loading efficiencies of nanoparticles into the cells is seen, with little effect on cell morphology even following extended nanoparticle exposure (up to 72 h). Nanoparticle incubation time and the fast response of the nanosensor (-2 s) make it a very powerful tool in monitoring the processes occurring within the cytosol.

Episodic ataxias (EAs) are rare neurological channelopathies that are characterized by spells of imbalance and a lack of co-ordination. There are seven clinically recognized EAs and multiple isolated cases. Five disease-causing genes have been identified to date. We describe a novel form of autosomal dominant EA in a large three-generation Irish family. This form of EA presents in early childhood with periods of unsteadiness generalized weakness and slurred speech during an attack, which may be triggered by physical tiredness or stress. Linkage analysis undertaken in 13 related individuals identified a single disease locus (1p36.13-p34.3) with a LOD score of 3.29. Exome sequencing was performed. Following data analysis, which included presence/absence within the linkage peak, two candidate variants were identified. These are located in the HSPG2 and UBR4 genes. UBR4 is an ubiquitin ligase protein that is known to interact with calmodulin, a Ca(2+) protein, in the cytoplasm. It also co-localizes with ITPR1 a calcium release channel that is a major determinant of mammal co-ordination. Although UBR4 is not an ion channel gene, the potential for disrupted Ca(2+) control within neuronal cells highlights its potential for a role in this form of EA.

There is increasing interest in the use of nanoparticles (NPs) for biomedical applications. In particular, nanobiophotonic approaches using fluorescence offers the potential of high sensitivity and selectivity in applications such as cell imaging and intracellular sensing. In this review, we focus primarily on the use of fluorescent silica NPs for these applications and, in so doing, aim to enhance and complement the key recent review articles on these topics. We summarize the main synthetic approaches, namely the Stöber and microemulsion processes, and, in this context, we deal with issues in relation to both covalent and physical incorporation of different types of dyes in the particles. The important issue of NP functionalization for conjugation to biomolecules is discussed and strategies published in the recent literature are highlighted and evaluated. We cite recent examples of the use of fluorescent silica NPs for cell imaging in the areas of cancer, stem cell and infectious disease research, and we review the current literature on the use of silica NPs for intracellular sensing of oxygen, pH and ionic species. We include a short final section which seeks to identify the main challenges and obstacles in relation to the potential widespread use of these particles for in vivo diagnostics and therapeutics.

Critical limitation of nanoparticles (NP) is their aggregation after functionalisation and antibody cross-linking. We analysed the cause of this aggregation with respect to functionalities (carboxyls and amines) on the NP surface. We have devised a low cost novel method to reduce such aggregations during protein cross-linking and validated it by probing the platelet surface with platelet surface-specific anti-CD41 antibody conjugated NPs.

We present a polymer biochip with embedded optics which allows the detection of supercritical angle fluorescence (SAF) without losses due to total internal reflection within the substrate. The chip design comprises structured spherical and aspherical optical elements on the bottom, while the top is chemically functionalized for direct binding of biomolecules. Furthermore, this design facilitates integration in lab-on-a-chip systems with appropriate microfluidics. In the confocal optical setup an ellipsoidal mirror is used for collection of SAF light above the critical angle of the water-polymer interface, which is detected by a photon-counting detector. The work presented here represents a proof of concept for performing sensitive and rapid point-of-care testing, using this low-cost, robust and disposable optical biochip platform. The performance of the platform was validated using direct binding DNA and human IgG assays which yielded low limits of detection 10 pM for DNA and 10 pg/ml for human IgG.

This paper describes the synthesis and characterization of sol-gel silica nanoparticles (NPs) derived from tetraethoxysilane (TEOS) and from tetraethoxysilane and methyltriethoxysilane (TEOS-MTEOS) in which is encapsulated, an in-house synthesized, stable oxygen-sensitive ruthenium complex, ruthenium (II) (bis-2,2-bipyridyl)-2(4-carboxylphenyl) imidazo[4,5-f][1,10]phenanthroline. These NPs were characterized using dynamic light scattering, transmission electron microscopy, scanning electron microscopy, Fourier transform infrared spectroscopy and Brunauer-Emmett-Teller analysis. The spherical, stable and monodispersed NPs have been prepared using the Stöber method. It was found that the addition of prehydrolyzed MTEOS-based sol prepared in an acidic environment to the reaction mixture containing TEOS NPs synthesized for 6 h produced material with increased porosity when compared to pure silica NPs. Oxygen sensitivity, stability, photobleaching and leaching have been characterized. The hybrid NPs exhibit enhanced O₂ sensitivity but a high degree of leaching when compared to pure silica NPs, which have minimum O₂ sensitivity and no leaching.

Platelets are central in maintaining normal haemostasis and are responsible for the cessation of blood loss following vascular injury. Platelet adhesion, leading to thrombus formation, involves rapid and distinct morphological changes culminating in cell aggregation. Rapid and highly specific platelet diagnostic tests are currently being developed to enable monitoring of drug response in patients with cardiovascular diseases. For a diagnostic device to be effective it needs to be rapid and simple, requiring minimal user interaction and providing results with minimal delay. This study describes the development of a rapid, label free platform for assay platelet function and demonstrates its use to monitor the influence of anti-thrombotic drugs. A rapid, single-step cell purification surface immobilises platelets from whole blood onto a protein array at specific locations used to define the assay site. Morphological changes and cell aggregation properties of activated platelets are exploited and combined within a label free, rapid, dye displacement chamber to determine cell morphology. Stimulation-dependent changes in cell morphology are described using both high resolution AFM imaging and a dye displacement platform.

In this paper we report a centrifugal microfluidic "lab-on-a-disc" system for at-line monitoring of human immunoglobulin G (hIgG) in a typical bioprocess environment. The novelty of this device is the combination of a heterogeneous sandwich immunoassay on a serial siphon-enabled microfluidic disc with automated sequential reagent delivery and surface-confined supercritical angle fluorescence (SAF)-based detection. The device, which is compact, easy-to-use and inexpensive, enables rapid detection of hIgG from a bioprocess sample. This was achieved with, an injection moulded SAF lens that was functionalized with aminopropyltriethoxysilane (APTES) using plasma enhanced chemical vapour deposition (PECVD) for the immobilization of protein A, and a hybrid integration with a microfluidic disc substrate. Advanced flow control, including the time-sequenced release of on-board liquid reagents, was implemented by serial siphoning with ancillary capillary stops. The concentration of surfactant in each assay reagent was optimized to ensure proper functioning of the siphon-based flow control. The entire automated microfluidic assay process is completed in less than 30 min. The developed prototype system was used to accurately measure industrial bioprocess samples that contained 10 mg mL(-1) of hIgG., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Nanoparticles are increasingly used as diagnostic tools due to the ease with which their surface chemistry, optical and physical properties can be controlled. Molecules, drugs, enzymes and fluorophores can be protected within the particle core or conjugated externally conferring nanoparticle biocompatibility, target specificity or environmental sensitivity. This study details the development and characterisation of stable, bright, dye-doped silica nanoparticles which are surface functionalised with PAMAM dendrimers to enable efficient conjugation to platelet activation-specific antibodies. We present the physical and optical properties and demonstrate colloidal stability. We also provide the first evidence of how NPs can be employed to specifically label human platelets immobilised on a lab-on-a-chip platform. Using a single step protocol, we demonstrate highly specific platelet labelling with the distribution of antibody-conjugated NPs matching that expected for the platelet GPIIb/IIIa receptor. The work highlights the potential of functionalized fluorescent NPs as diagnostic tools for cardiovascular disease., From the Clinical Editor: This study details the development and characterization of PAMAM dendrimer functionalized, stable, and bright dye-doped silica nanoparticles that enable efficient conjugation to platelet activation-specific antibodies. These fluorescent NPs may specifically label human platelets that can be used as diagnostic tools for cardiovascular disease., (Copyright © 2013 Elsevier Inc. All rights reserved.)

A fourth generation PAMAM dendrimer has been successfully employed for the development of a single step synthesis strategy for self-assembled Ag-Au nanohybrid structures. The surface plasmon resonance properties and the degree of self-assembly of the nanohybrid are strongly correlated with the stoichiometry of the metals which gives rise to enhanced plasmonic properties. The enhanced plasmonic response of the nanohybrids is modeled and is validated experimentally in a model HRP (horseradish peroxidise) bioassay carried out on an SPR-based biochip platform.

Fluorescent nanoparticle (NP) labels are of great interest for point-of-care medical diagnostics where high fluorescence signals combined with low limits of detection are required. In this work, hydrophilic and hydrophobic fluorescein dye derivatives were covalently doped into silica NPs. The NPs were prepared in a range of sizes from 16 to 80 nm using both ternary and quaternary microemulsion methods where the diameter varied linearly with changes in the water to surfactant ratio. The morphology and colloidal stability of the NPs were characterised using transmission electron microscopy and photon correlation spectroscopy; NPs doped with hydrophobic fluorescein dye were significantly smaller and more polydispersed. Optical properties including absorption, fluorescence and quantum efficiency were also determined. Representative NPs from each microemulsion method (ternary, Ø = 25 nm and quaternary, Ø = 80 nm) were tested as labels in a fluorescence based immunoassay for the detection of human IgG and human chorionic gonadotropin. Both sets of nanoparticle assays showed lower limits of detection and better coefficients of variance than a free dye label with good day to day reproducibility. The optimal surface coverage of detection antibody was also found to depend on the size of the nanoparticle.

In this work, gold-silica plasmonic nanohybrids have been synthesized as model systems which enable tuning of dye fluorescence enhancement/quenching interactions. For each system, a dye-doped silica core is surrounded by a 15 nm spacer region, which in turn is surrounded by gold nanoparticles (GNPs). The GNPs are either covalently conjugated via mercapto silanization to the spacer or encapsulated in a separate external silica shell. The intermediate spacer region can be either dye doped or left undoped to enable quenching and plasmonic enhancement effects respectively. The study indicates that there is a larger enhancement effect when GNPs are encapsulated in the outer shell compared to the system of external conjugation. This is due to the environmental shielding provided by shell encapsulation compared to the exposure of the GNPs to the solvent environment for the externally conjugated system. The fluorescence signal enhancement of the nanohybrid systems was evaluated using a standard HRP-anti-HRP fluorescence based assay platform.

A highly bright and photostable, fluorescent nanohybrid particle is presented which consists of gold nanoparticles (GNPs) embedded in dye-doped silica in a core-shell configuration. The dye used is the near-infrared emitting 4,5-benzo-5'-(iodoacetaminomethyl)-1',3,3,3',3'-pentamethyl-1-(4-sulfobutyl) indodicarbo cyanine. The nanohybrid architecture comprises a GNP core which is separated from a layer of dye molecules by a 15 nm buffer layer and has an outer protective, undoped silica shell. Using this architecture, a brightness factor of 550 has been achieved compared to the free dye. This hybrid system, referred to as Noble metal Enhanced Optical Nanohybrid (NEON) in this paper, is the first nanohybrid construct to our knowledge which demonstrates such tunable fluorescence property. NEON has enhanced photostability compared to the free dye and compared to a control particle without GNPs. Furthermore, the NEON particle, when used as a fluorescent label in a model bioassay, shows improved performance over assays using a conventional single dye molecule label.

Background: Neuroblastoma is one of the most challenging malignancies of childhood, being associated with the highest death rate in paediatric oncology, underlining the need for novel therapeutic approaches. Typically, patients with high risk disease undergo an initial remission in response to treatment, followed by disease recurrence that has become refractory to further treatment. Here, we demonstrate the first silica nanoparticle-based targeted delivery of a tumor suppressive, pro-apoptotic microRNA, miR-34a, to neuroblastoma tumors in a murine orthotopic xenograft model. These tumors express high levels of the cell surface antigen disialoganglioside GD2 (GD(2)), providing a target for tumor-specific delivery., Principal Findings: Nanoparticles encapsulating miR-34a and conjugated to a GD(2) antibody facilitated tumor-specific delivery following systemic administration into tumor bearing mice, resulted in significantly decreased tumor growth, increased apoptosis and a reduction in vascularisation. We further demonstrate a novel, multi-step molecular mechanism by which miR-34a leads to increased levels of the tissue inhibitor metallopeptidase 2 precursor (TIMP2) protein, accounting for the highly reduced vascularisation noted in miR-34a-treated tumors., Significance: These novel findings highlight the potential of anti-GD(2)-nanoparticle-mediated targeted delivery of miR-34a for both the treatment of GD(2)-expressing tumors, and as a basic discovery tool for elucidating biological effects of novel miRNAs on tumor growth.

Fluorescence lifetime correlation spectroscopy (FLCS) is presented as a single-step label-free detection method for probing the amine silanization-driven spontaneous 3D self-assembly of freestanding gold nanoparticles (GNPs) in solution. Unlike the conventional methods of studying self-assembly, for example, UV-vis spectroscopy and electron microscopy, FLCS utilizes the intrinsic gold fluorescence. The significance of this approach is to amalgamate the measurement of optical and hydrodynamic size properties simultaneously to achieve a more coherent description of the self-assembly pathway. GNP self-assembly has two-stage kinetics. Electrostatic interaction drives the initial amine silanization, and this is followed by siloxane bond formation between hydrolyzed ethoxy groups of GNP-attached APTES, resulting in the formation of micrometer-sized superstructures. The self-assembly has resulted in a 5-fold increase in the fluorescence lifetime (FL), and the FLCS study has shown an 8- to 10-fold increase in the diffusion coefficient using the pure diffusion model. This result is consistent with the transmission electron microscopy (TEM) observation, which shows a few hundred fold increase in the diameter due to assembly formation by the GNPs., (© 2011 American Chemical Society)

This article describes the synthesis and characterisation of fluorescent composite nanoparticles consisting of a silica core and a dextran shell. The silica core contains a rhodamine-based reference dye, which allows ratiometric measurements and the dextran shell is labelled with the Ca(2+)-sensitive dye Fluo-4. The nanoparticles have an average hydrodynamic diameter of 95 nm, good colloidal stability and show a 2.9-fold increase in fluorescence intensity upon binding to Ca(2+) ions. The apparent dissociation constant of K'(d) ≈ 520 nM is well suited for measurements in the physiological range.

We present a novel approach to the enhancement of surface plasmon-coupled emission (SPCE) using surface plasmon excitation in a bimetal (Ag/Au) layer and we validate the enhancement by presenting the results of a model human IgG immunoassay. Theoretical calculations using Fresnel's equations have been carried out to determine the optimum bimetallic composition and the resulting electric field enhancement. Signal enhancement of SPCE was confirmed using a range of bimetallic layers which were deposited on the surface of a high collection efficiency polymer array biochip and subsequently immobilized with Alexa Fluor 647 labeled anti-human IgG. The bimetallic film of Ag/Au (36/10nm) was determined as an optimum substrate for maximum SPCE signal which was a compromise between the long-term stability of the metal layer and the optimized evanescent field enhancement. An enhanced dose-dependent response was also demonstrated which was ∼3 times greater than that detected with a pure gold layer. A human IgG immunoassay showed a dose-dependent response yielding a limit of detection of 1pg/ml by the 3σ rule. The improved performance of the bimetal layer compared to that of an assay carried out on a pure gold layer is attributed to the enhanced evanescent field intensity of surface plasmons in the bimetal combination which excites more fluorescence hence producing an enhanced SPCE signal. This result demonstrates the potential of the SPCE-based array biochips as a sensitive and high-throughput analysis platform for biomolecular interactions., (Copyright © 2010 Elsevier B.V. All rights reserved.)

This Article addresses the important issue of the characterization of surface functional groups for optical bioassay applications. We use a model system consisting of spherical dye-doped silica nanoparticles (NPs) that have been functionalized with amine groups whereby the encapsulated cyanine-based near-infrared dye fluorescence acts as a probe of the NP surface environment. This facilitates the identification of the optimum deposition parameters for the formation of a stable ordered amine monolayer and also elucidates the functionalization profile of the amine-silanization process. Specifically, we use a novel approach where the techniques of fluorescence correlation spectroscopy (FCS) and fluorescence lifetime measurement (FL) are used in conjunction with the more conventional analytical techniques of zeta potential measurement and Fourier transfer infrared spectroscopy (FTIR). The dynamics of the ordering of the amine layer in different stages of the reaction have been characterized by FTIR, FL, and FCS. The results indicate an optimum reaction time for the formation of a stable amine layer, which is optimized for further biomolecular conjugation, whereas extended reaction times lead to a disordered cross-linked layer. The results have been validated using an immunoglobulin (IgG) plate-based direct binding assay where the maximum number of IgG-conjugated aminated NPs were captured by immobilized anti-IgG antibodies for the NP sample corresponding to the optimized amine-silanization condition. Importantly, these results point to the potential of FCS and FL as useful analytical tools in diverse fields such as characterization of surface functionalization.

The focus of this work is on the development and characterisation of a fluorescence-based ratiometric sol-gel-derived dissolved carbon dioxide (dCO(2)) sensor for use in environmental monitoring applications. Fluorescence-based dCO(2) sensors are attractive as they facilitate the development of portable and low-cost systems that can be easily deployed outside the laboratory environment. The sensor developed for this work exploits a pH fluorescent dye 1-hydroxypyrene-3,6,8-trisulfonic acid, ion-paired with cetyltrimethylammonium bromide (HPTS-IP), which has been entrapped in a hybrid sol-gel-based matrix derived from n-propyltriethoxysilane along with the liphophilic organic base. The sensor spot deposited on a cover slip has been interrogated with a robust, ratiometric optical probe that combines effective fluorescence excitation and detection and thus facilitates the production of a highly sensitive sensor system using low-cost optoelectronic components. The probe design involves the use of dual-LED excitation in order to facilitate ratiometric operation and uses a silicon PIN photodiode. HPTS-IP exhibits two pH-dependent changes in excitation bands, which allows for dual excitation ratiometric detection as an indirect measure of the dCO(2). Such measurements are insensitive to changes in dye concentration, leaching and photobleaching of the fluorophore and instrument fluctuations unlike unreferenced fluorescence intensity measurements. The performance of the sensor system is characterised by a high degree of repeatability, reversibility and stability. Calculated limit of detection for the sensor was 35 ppb. The sensor probe was used to monitor dCO(2) levels in a laboratory-based aquatic habitat, and the expected diurnal pattern was clearly visible. The influence of temperature, biofouling and photobleaching on sensor performance has been also investigated.

The technique of surface plasmon-coupled emission (SPCE) involves the coupling of light which is emitted from a fluorophore into the surface plasmon of an adjacent thin metal film, giving rise to highly directional emission. We have combined the advantages of SPCE with the high light collection efficiency of supercritical angle fluorescence by carrying out an immunoassay on a paraboloid array biochip in the absence of the conventional SPCE spacer layer normally used to minimize metal quenching of the fluorescence. In this work, we have successfully demonstrated an SPCE-based assay by utilizing the protein assay layer as the spacer layer. A novel 3 × 3 injection molded polymer biochip with paraboloid elements was used. The paraboloid elements served to enhance the light collection efficiency while the top surface was coated with a gold layer to use excitation of surface plasmons and detection of SPCE emission. Theoretical modeling of the gold-protein layer structure showed that the surface plasmon resonance angles were located in the detection range of the paraboloid biochip. The polarization dependence of SPCE emission was also demonstrated. Finally, a human IgG sandwich immunoassay was carried out which exhibited a limit of detection of ~10 ng/ml using 3σ. The results demonstrate the potential of the SPCE-based paraboloid array biochip as a novel platform for high-throughput analysis of biomolecular interactions.

Fluorescence lifetime (FL) analysis and fluorescence correlation spectroscopy (FCS) have been successfully employed to reveal detailed information about the internal architecture of fluorescent silica nanoparticles (NPs). The dual-component lifetime behavior shows a two-domain dye distribution in the NP as a function of solvent accessibility. The introduction of an undoped silica shell serves to stabilize the outer dye fraction that is manifest as an increase in lifetime. The FCS not only shows a size-dependent increase in the cross-correlation decay constant but also demonstrates a significant relaxation in the FCS signal with the introduction of an undoped silica shell.

This paper describes the fabrication and performance of a range of highly sensitive luminescence-based oxygen sensor films based on the fluorinated sol-gel precursor 3,3,3-trifluoropropyltrimethoxysilane (TFP-TMOS). The oxygen-sensitive ruthenium complex [Ruthenium(II)-tris(4,7-diphenyl-1,10-phenanthroline)] dichloride, [Ru(dpp)(3)](2+) was entrapped in a wide range of ORMOSILs (organically modified silicates) matrices composed of alkyl and TFP-TMOS sol-gel precursors in different relative ratios. The influence of TFP-TMOS on sensor sensitivity, humidity-sensitivity and long-term stability was investigated and performance was compared to that of similar but non-fluorinated films. The optimum limit of detection was found to be 0.002% of oxygen for the propyltriethoxysilane (PTEOS)/TFP-TMOS-derived film compared to 0.09% for PTEOS-derived films reported previously. Photobleaching of the luminescent complex in fluorinated and non-fluorinated matrices was also investigated. It was established that photobleaching was reduced but not eliminated in fluorinated films. All films produced in this study exhibit very good reproducibility, reversibility, enhanced sensitivity, humidity-insensitivity and long-term stability., (2010 Elsevier B.V. All rights reserved.)

We have carried out a human IgG immunoassay on a novel disposable optical array biochip using surface plasmon-coupled emission (SPCE) detection. The work successfully combines the advantages of the highly directional SPCE emission profile and enhanced surface plasmon excitation with the high light collection efficiency achieved using supercritical angle fluorescence (SAF). This is achieved using an array of transparent paraboloid polymer elements which have been coated with a thin gold layer to facilitate SPCE. Moreover, since only the emission of molecules which are close to the metal surface couple into the surface plasmon, the detection is highly surface-specific leading to background suppression and increased signal-to-noise ratio. Theoretical calculations have been carried out in order to match the surface plasmon resonance angles and SPCE emission angles to the paraboloid array features for light collection. A sandwich assay format was used and a dose response curve was obtained in the concentration range 2 ng/ml to 200 microg/ml yielding a limit of detection of 20 ng/ml. This is the first demonstration of an SPCE-based assay on a disposable biochip platform and indicates the potential of SPCE-based arrays for high-throughput analysis of biomolecular interactions., (Crown Copyright 2009. Published by Elsevier B.V. All rights reserved.)

In this work, we used a model assay system (polyclonal human IgG-goat antihuman IgG) to elucidate some of the key factors that influence the analytical performance of bioassays that employ metal-enhanced fluorescence (MEF) using silver nanoparticles (NPs). Cy5 dye was used as the fluorescent label, and results were compared with a standard assay performed in the absence of NPs. Two sizes of silver NPs were prepared with respective diameters of 60 +/- 10 and 149 +/- 16 nm. The absorption spectra of the NPs in solution were fitted accurately using Mie theory, and the dipole resonance of the 149-nm NPs in solution was found to match well with the absorption spectrum of Cy5. Such spectral matching is a key factor in optimizing MEF. NPs were deposited uniformly and reproducibly on polyelectrolyte-coated polystyrene substrates. Compared to the standard assay performed without the aid of NPs, significant improvements in sensitivity and in limit of detection (LOD) were obtained for the assay with the 149-nm NPs. An important observation was that the relative enhancement of fluorescence increased as the concentration of antigen increased. The metal-assisted assay data were analyzed using standard statistical methods and yielded a LOD of 0.086 ng/mL for the spectrally matched NPs compared to a value of 5.67 ng/mL obtained for the same assay in the absence of NPs. This improvement of approximately 66x in LOD demonstrates the potential of metal-enhanced fluorescence for improving the analytical performance of bioassays when care is taken to optimize the key determining parameters.

In this paper, we describe a novel technique for depositing metal nanoparticles (NPs) on a planar substrate whereby the NPs are micro-patterned on the surface by a simple stamp-printing procedure. The method exploits the attractive force between negatively charged colloidal metal NPs and positively-charged polyelectrolyte layers which have been selectively deposited on the surface. Using this technique, large uniform areas of patterned metal NPs, with different plasmonic properties, were achieved by optimisation of the stamping process. We report the observation of unusual fluorescence emission from these structures. The emission was measured using epifluorescence microscopy. Fluorescence lifetime behaviour was also measured. Furthermore, the mu-patterned NPs exhibited blinking behaviour under 469 nm excitation and the fluorescence spectrum was multi-peaked. It has been established that the fluorescence is independent of the plasmon resonance properties of the NPs. As well as optimising the novel NP mu-patterning technique, this work discusses the origin and characteristics of the anomalous fluorescence behaviour in order to characterise and minimise this unwanted background contribution in the use of metal NPs for plasmonic enhancement of fluorescence for optical biochip applications.