Spatial-Division Multiplexing Approach for Simultaneous Detection of Fiber-Optic Ball Resonator Sensors: Applications for Refractometers and Biosensors.

Fiber-optic ball resonators are an attractive technology for refractive index (RI) sensing and optical biosensing, as they have good sensitivity and allow for a rapid and repeatable manufacturing process. An important feature for modern biosensing devices is the multiplexing capacity, which allows for interrogating multiple sensors (potentially, with different functionalization methods) simultaneously, by a single analyzer. In this work, we report a multiplexing method for ball resonators, which is based on a spatial-division multiplexing approach. The method is validated on four ball resonator devices, experimentally evaluating both the cross-talk and the spectral shape influence of one sensor on another. We show that the multiplexing approach is highly efficient and that a sensing network with an arbitrary number of ball resonators can be designed with reasonable penalties for the sensing capabilities. Furthermore, we validate this concept in a four-sensor multiplexing configuration, for the simultaneous detection of two different cancer biomarkers across a widespread range of concentrations.

Cost-Effective Fiber Optic Solutions for Biosensing.

In the last years, optical fiber sensors have proven to be a reliable and versatile biosensing tool. Optical fiber biosensors (OFBs) are analytical devices that use optical fibers as transducers, with the advantages of being easily coated and biofunctionalized, allowing the monitorization of all functionalization and detection in real-time, as well as being small in size and geometrically flexible, thus allowing device miniaturization and portability for point-of-care (POC) testing. Knowing the potential of such biosensing tools, this paper reviews the reported OFBs which are, at the moment, the most cost-effective. Different fiber configurations are highlighted, namely, end-face reflected, unclad, D- and U-shaped, tips, ball resonators, tapered, light-diffusing, and specialty fibers. Packaging techniques to enhance OFBs' application in the medical field, namely for implementing in subcutaneous, percutaneous, and endoscopic operations as well as in wearable structures, are presented and discussed. Interrogation approaches of OFBs using smartphones' hardware are a great way to obtain cost-effective sensing approaches. In this review paper, different architectures of such interrogation methods and their respective applications are presented. Finally, the application of OFBs in monitoring three crucial fields of human life and wellbeing are reported: detection of cancer biomarkers, detection of cardiovascular biomarkers, and environmental monitoring.

Ultra-wide, attomolar-level limit detection of CD44 biomarker with a silanized optical fiber biosensor.

Measuring cancer biomarkers at ultralow detection limit and high sensitivity could be a promising tool for early diagnosis, monitoring treatment and post-treatment recurrence. Soluble CD44 is a promising diagnostic and prognostic biomarker in several types of cancer including gastric, colon and breast cancer. Several highly sensitive biosensors have been built to measure this important biomarker. However, they did not reach attomolar level of detection. The aim of this work was to build a biosensor capable of detecting CD44 concentrations down to attomolar (aM) level while measuring it in a wide concentration range. Herein, we demonstrate a biosensor that offers 4 key advantages over existing platforms for CD44 detection: 1) detection of CD44 was carried out in a diluted serum down to attomolar level (4.68 aM) which is about 6 orders of magnitude lower than that of a traditional ELISA; 2) fabrication of the sensor is done in a fast way using inexpensive materials making it a disposable fiber optic biosensor; 3) detection of CD44 was performed in a wide dynamic range previously not shown in other similar biosensors; 4) a proof-of-concept experiment was performed using the biosensor to embed it in a catheter to measure the protein in flow conditions.

Label-free fiber-optic spherical tip biosensor to enable picomolar-level detection of CD44 protein.

Increased level of CD44 protein in serum is observed in several cancers and is associated with tumor burden and metastasis. Current clinically used detection methods of this protein are time-consuming and use labeled reagents for analysis. Therefore exploring new label-free and fast methods for its quantification including its detection in situ is of importance. This study reports the first optical fiber biosensor for CD44 protein detection, based on a spherical fiber optic tip device. The sensor is easily fabricated from an inexpensive material (single-mode fiber widely used in telecommunication) in a fast and robust manner through a CO2 laser splicer. The fabricated sensor responded to refractive index change with a sensitivity of 95.76 dB/RIU. The spherical tip was further functionalized with anti-CD44 antibodies to develop a biosensor and each step of functionalization was verified by an atomic force microscope. The biosensor detected a target of interest with an achieved limit of detection of 17 pM with only minor signal change to two control proteins. Most importantly, concentrations tested in this work are very broad and are within the clinically relevant concentration range. Moreover, the configuration of the proposed biosensor allows its potential incorporation into an in situ system for quantitative detection of this biomarker in a clinical setting.

Optical Fiber Ball Resonator Sensor Spectral Interrogation through Undersampled KLT: Application to Refractive Index Sensing and Cancer Biomarker Biosensing.

Optical fiber ball resonators based on single-mode fibers in the infrared range are an emerging technology for refractive index sensing and biosensing. These devices are easy and rapid to fabricate using a CO2 laser splicer and yield a very low finesse reflection spectrum with a quasi-random pattern. In addition, they can be functionalized for biosensing by using a thin-film sputtering method. A common problem of this type of device is that the spectral response is substantially unknown, and poorly correlated with the size and shape of the spherical device. In this work, we propose a detection method based on Karhunen-Loeve transform (KLT), applied to the undersampled spectrum measured by an optical backscatter reflectometer. We show that this method correctly detects the response of the ball resonator in any working condition, without prior knowledge of the sensor under interrogation. First, this method for refractive index sensing of a gold-coated resonator is applied, showing 1594 RIU-1 sensitivity; then, this concept is extended to a biofunctionalized ball resonator, detecting CD44 cancer biomarker concentration with a picomolar-level limit of detection (19.7 pM) and high specificity (30-41%).

Shallow-Tapered Chirped Fiber Bragg Grating Sensors for Dual Refractive Index and Temperature Sensing.

In this work, we present a gold-coated shallow-tapered chirped fiber Bragg grating (stCFBG) for dual refractive index (RI) and temperature sensing. The stCFBG has been fabricated on a 15-mm long chirped FBG, by tapering a 7.29-mm region with a waist of 39 µm. The spectral analysis shows two distinct regions: a pre-taper region, in which the stCFBG is RI-independent and can be used to detect thermal changes, and a post-taper region, in which the reflectivity increases significantly when the RI increments. We estimate the RI and thermal sensitivities as 382.83 dB/RIU and 9.893 pm/°C, respectively. The cross-talk values are low (-1.54 × 10-3 dB/°C and 568.1 pm/RIU), which allows an almost ideal separation between RI and thermal characteristics. The stCFBG is a compact probe, suitable for long-term and temperature-compensated biosensing and detection of chemical analytes.

Fiber Optic Refractive Index Sensors Based on a Ball Resonator and Optical Backscatter Interrogation.

In this work, we introduced fabrication and interrogation of simple and highly sensitive fiber-optic refractive index (RI) sensors based on ball resonators built on the tip of single-mode fibers. The probes have been fabricated through a CO2 fiber splicer, with a fast (~600 s) and repeatable method. The ball resonator acted as a weak interferometer with a return loss below -50 dB and was interrogated with an optical backscatter reflectometer measuring the reflection spectrum. The ball resonators behaved as weak interferometers with a shallow fringe and a spectrum that appeared close to a random signal, and RI sensitivity could be measured either through wavelength shift or amplitude change. In this work, we reported four samples having sensitivity ranges 48.9-403.3 nm/RIU and 256.0-566.2 dB/RIU (RIU = refractive index unit). Ball resonators appeared as a sensitive and robust platform for RI sensing in liquid and can be further functionalized for biosensing.

Functionalized etched tilted fiber Bragg grating aptasensor for label-free protein detection.

An aptasensor based on etched tilted fiber Bragg grating (eTFBG) is developed on a single-mode optical fiber targeting biomolecule detection. TFBGs were chemically etched using hydrofluoric acid (HF) to partially remove the fiber cladding. The sensor response was coarsely interrogated, resulting on a sensitivity increase from 1.25 nm/RIU (refractive index unit) at the beginning of the process, up to 23.38 nm/RIU at the end of the etching, for a RI range from 1.3418 to 1.4419 RIU. The proposed aptasensor showed improved RI sensitivity as compared to the unetched TFBG, without requiring metal depositions on the fiber surface or polarization control during the measurements. The proposed sensor was tested for the detection of thrombin-aptamer interactions based on silane-coupling surface chemistry, with thrombin concentrations ranging from 2.5 to 40 nM. Functionalized eTFBGs provided a competitive platform for biochemical interaction measurements, showing sensitivity values ranging from 2.3 to 3.3 p.m./nM for the particular case of thrombin detection.

Wavelet-Based Demodulation of Multimode Etched Fiber Bragg Grating Refractive Index Sensor.

Etched fiber Bragg grating (EFBG)-based sensors are used as evanescent field sensors for refractive index detection. When the fiber thickness is thin and the refractive index sensitivity increases, the number of propagating modes increases, resulting in a spectral enlargement that complicates the interrogation of the sensor. In this work, we present a method to analyze the spectrum of a multimode etched fiber Bragg grating (MMEFBG) in the wavelet domain, which analyzes the amount of spectral density independently from the peak reflectivity value. The proposed interrogation method permits defining the integral of the spectral density as a novel and unconventional estimator. With respect to the conventional estimators based on wavelength shift, this estimator can better exploit the larger amount of information given by the spectral enlargement typical of multimode behavior. Results were obtained by etching an MMEFBG in hydrofluoric acid and using water/sucrose mixtures to evaluate the refractive index sensitivity, validating the interrogation method.

Fiber optic refractive index sensors through spectral detection of Rayleigh backscattering in a chemically etched MgO-based nanoparticle-doped fiber.

We demonstrate and experimentally validate a fiber optic refractive index (RI) sensor obtained by simply etching a high-scattering MgO-based nanoparticle-doped single-mode fiber in hydrofluoric acid (HF). The fiber has 32.3 dB stronger Rayleigh scattering than a standard fiber, allowing a detection of scattering spectral signatures with an optical backscatter reflectometer, even when the core is exposed to the outer RI. The obtained sensitivity is 1.53 nm/RIU (RI units), measured by correlating the scattering spectra. We prove the possibility of implementing a distributed RI detection (seven locations spaced by 1 mm). The fabrication method for this RI sensor is simplified, since it simply requires etching in an HF bath, without the need of inscribing reflective elements or fabricating microstructures in the fiber.

Etched Fiber Bragg Grating Biosensor Functionalized with Aptamers for Detection of Thrombin.

A biosensor based on an etched Fiber Bragg Grating (EFBG) for thrombin detection is reported. The sensing system is based on a Fiber Bragg Grating (FBG) with a Bragg wavelength of 1550 nm, wet-etched in hydrofluoric acid (HF) for ~27 min, to achieve sensitivity to a refractive index (RI) of 17.4 nm/RIU (refractive index unit). Subsequently, in order to perform a selective detection of thrombin, the EFBG has been functionalized with silane-coupling agent 3-(aminopropyl)triethoxysilane (APTES) and a cross-linker, glutaraldehyde, for the immobilization of thrombin-binding aptamer. The biosensor has been validated for thrombin detection in concentrations ranging from 10 nM to 80 nM. The proposed sensor presents advantages with respect to other sensor configurations, based on plasmonic resonant tilted FBG or Long Period Grating (LPG), for thrombin detection. Firstly, fabricating an EFBG only requires chemical etching. Moreover, the functionalization method used in this study (silanization) allows the avoidance of complicated and expensive fabrications, such as thin film sputtering or chemical vapor deposition. Due to their characteristics, EFBG sensors are easier to multiplex and can be used in vivo. This opens new possibilities for the detection of thrombin in clinical settings.

Fabrication and Interrogation of Refractive Index Biosensors Based on Etched Fiber Bragg Grating (EFBG).

We discuss the fabrication and interrogation of a fiber-optic refractive index sensors based on wet-etched fiber Bragg grating (EFBG). The fabrication is based on chemical etching of an FBG through an HF-free etching solution (ammonium fluoride and sulfuric acid), which progressively depletes the fiber cladding exposing the core to the outer medium. Microscope inspection of the fiber and real-time detection of the Bragg wavelength allow controlling the sensitivity. The proposed interrogation method is based on a spline interpolation, that measures the change of Bragg wavelength when the FBG is exposed to variations of the refractive index in the surrounding medium. An experimental validation has been carried out, for small refractive index variations (up to $1.85\times 10^{-3}$ RIU), in order to verify the progressive change of sensitivity through fiber etching. The proposed EFBG sensing unit is a building block for functionalized fiber optic biosensors.

Partially etched chirped fiber Bragg grating (pECFBG) for joint temperature, thermal profile, and refractive index detection.

In this work, a partially etched chirped fiber Bragg grating (pECFBG) is introduced, as a compact sensor for multi-parametric measurement of temperature, thermal gradients over the active length, and refractive index. The sensor is fabricated by wet-etching a portion of a 14-mm linearly chirped FBG with linear chirp profile. The resulting device has two active areas: the unetched part of the grating (2 mm) can be used either as a uniform temperature sensor, or to detect thermal gradients experienced through the grating length by means of a spectral reconstruction technique; the etched part (12 mm), besides having a similar thermal sensitivity, is exposed to refractive index sensing through the introduction of a sensitivity to external refractive index. Overall, the pECFBG structure behaves as a compact sensor with multi-parameter capability, that can both measure temperature and refractive index on the same grating, but also spatially resolve temperature detection through the grating section. The results have been validated through both a model and experimental setup, showing that the mutual correlation algorithm applied to different spectral parts of the grating is able to discriminate between uniform and gradient-shaped temperature profiles, and refractive index changes.

Biocompatible scaffolds based on natural polymers for regenerative medicine.

The chitosan and gelatine are commonly used biopolymers for the tissue engineering applications. In the previous methods for the cryogels synthesis, multistep preparation methods using toxic cross-linking agents such as glutaraldehyde are reported. Here, we present a two-step preparation method of gelatin macroporous cryogels and one-step preparation method of chitosan or gelatin cryogels. The physico-chemical properties of obtained scaffolds were characterized using FTIR, zeta potential, SEM and laser confocal microscopy. Non-toxic and biodegradable cross-linking agents such as oxidized dextran and 1,1,3,3-tetramethoxypropane are utilized. The one-step chitosan cryogels had degradation degree ~2 times higher compared to the cryogels prepared with a two-step method i.e. reduced by borohydride. Scaffolds cross-linked by glutaraldehyde had about 40% viability, whereas nine various compositions of cryogels showed significantly higher viability (~80%) of fibroblast cells in vitro. The cryogels were obtained without using the harmful compounds and therefore can be used straightforward as biocompatible and biodegradable scaffolds for the cell culturing purposes and other biomedical applications.

The effects of antiviral treatment on breast cancer cell line.

BACKGROUND: Recent studies have revealed the positive antiproliferative and cytotoxic effects of antiviral agents in cancer treatment. The real effect of adjuvant antiviral therapy is still controversial due to the lack of studies in biochemical mechanisms. Here, we studied the effect of the antiviral agent acyclovir on morphometric and migratory features of the MCF7 breast cancer cell line. Molecular levels of various proteins have also been examined. METHODS: To evaluate and assess the effect of antiviral treatment on morphometric, migratory and other cellular characteristics of MCF7 breast cancer cells, the following experiments were performed: (i) MTT assay to measure the viability of MCF7 cells; (ii) Colony formation ability by soft agar assay; (iii) Morphometric characterization by immunofluorescent analysis using confocal microscopy; (iv) wound healing and transwell membrane assays to evaluate migration and invasion capacity of the cells; (v) ELISA colorimetric assays to assess expression levels of caspase-3, E-cadherin and enzymatic activity of aldehyde dehydrogenase (ALDH). RESULTS: We demonstrate the suppressive effect of acyclovir on breast cancer cells. Acyclovir treatment decreases the growth and the proliferation rate of cells and correlates with the upregulated levels of apoptosis associated cytokine Caspase-3. Moreover, acyclovir inhibits colony formation ability and cell invasion capacity of the cancer cells while enhancing the expression of E-cadherin protein in MCF7 cells. Breast cancer cells are characterized by high ALDH activity and associated with upregulated proliferation and invasion. According to this study, acyclovir downregulates ALDH activity in MCF7 cells. CONCLUSIONS: These results are encouraging and demonstrate the possibility of partial suppression of cancer cell proliferation using an antiviral agent. Acyclovir antiviral agents have a great potential as an adjuvant therapy in the cancer treatment. However, more research is necessary to identify relevant biochemical mechanisms by which acyclovir induces a potent anti-cancer effect.

Disruption of Bcl-2 and Bcl-xL by viral proteins as a possible cause of cancer.

The Bcl proteins play a critical role in apoptosis, as mutations in family members interfere with normal programmed cell death. Such events can cause cell transformation, potentially leading to cancer. Recent discoveries indicate that some viral proteins interfere with Bcl proteins either directly or indirectly; however, these data have not been systematically described. Some viruses encode proteins that reprogramme host cellular signalling pathways controlling cell differentiation, proliferation, genomic integrity, cell death, and immune system recognition. This review analyses and summarises the existing data and discusses how viral proteins interfere with normal pro- and anti-apoptotic functions of Bcl-2 and Bcl-xL. Particularly, this article focuses on how viral proteins, such as Herpesviruses, HTLV-1, HPV and HCV, block apoptosis and how accumulation of such interference predisposes cancer development. Finally, we discuss possible ways to prevent and treat cancers using a combination of traditional therapies and antiviral preparations that are effective against these viruses.