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Publicações

Publicações por CAP

2018

Alternative SNP detection platforms, HRM and biosensors, for varietal identification in Vitis vinifera L. using F3H and LDOX genes

Autores
Gomes, S; Castro, C; Barrias, S; Pereira, L; Jorge, P; Fernandes, JR; Martins Lopes, P;

Publicação
SCIENTIFIC REPORTS

Abstract
The wine sector requires quick and reliable methods for Vitis vinifera L. varietal identification. The number of V. vinifera varieties is estimated in about 5,000 worldwide. Single Nucleotide Polymorphisms (SNPs) represent the most basic and abundant form of genetic sequence variation, being adequate for varietal discrimination. The aim of this work was to develop DNA-based assays suitable to detect SNP variation in V. vinifera, allowing varietal discrimination. Genotyping by sequencing allowed the detection of eleven SNPs on two genes of the anthocyanin pathway, the flavanone 3-hydroxylase (F3H, EC: 1.14.11.9), and the leucoanthocyanidin dioxygenase (LDOX, EC 1.14.11.19; synonym anthocyanidin synthase, ANS) in twenty V. vinifera varieties. Three High Resolution Melting (HRM) assays were designed based on the sequencing information, discriminating five of the 20 varieties: Alicante Bouschet, Donzelinho Tinto, Merlot, Moscatel Galego and Tinta Roriz. Sanger sequencing of the HRM assay products confirmed the HRM profiles. Three probes, with different lengths and sequences, were used as bio-recognition elements in an optical biosensor platform based on a long period grating (LPG) fiber optic sensor. The label free platform detected a difference of a single SNP using genomic DNA samples. The two different platforms were successfully applied for grapevine varietal identification.

2018

Plasmonic Optical Fiber Sensor Based on Double Step Growth of Gold Nano-Islands

Autores
de Almeida, JMMM; Vasconcelos, H; Jorge, PAS; Coelho, L;

Publicação
SENSORS

Abstract
It is presented the fabrication and characterization of optical fiber sensors for refractive index measurement based on localized surface plasmon resonance (LSPR) with gold nano-islands obtained by single and by repeated thermal dewetting of gold thin films. Thin films of gold deposited on silica (SiO2) substrates and produced by different experimental conditions were analyzed by Scanning Electron Microscope/Dispersive X-ray Spectroscopy (SEM/EDS) and optical means, allowing identifying and characterizing the formation of nano-islands. The wavelength shift sensitivity to the surrounding refractive index of sensors produced by single and by repeated dewetting is compared. While for the single step dewetting, a wavelength shift sensitivity of similar to 60 nm/RIU was calculated, for the repeated dewetting, a value of similar to 186 nm/RIU was obtained, an increase of more than three times. It is expected that through changing the fabrication parameters and using other fiber sensor geometries, higher sensitivities may be achieved, allowing, in addition, for the possibility of tuning the plasmonic frequency.

2018

Experimental and Theoretical Evaluation of the Trapping Performance of Polymeric Lensed Optical Fibers: Single Biological Cells versus Synthetic Structures

Autores
Paiva, JS; Ribeiro, RSR; Jorge, PAS; Rosa, CC; Azevedo, MM; Sampaio, P; Cunha, JPS;

Publicação
BIOPHOTONICS: PHOTONIC SOLUTIONS FOR BETTER HEALTH CARE VI

Abstract
Optical Tweezers (OTs) have been widely applied in Biology, due to their outstanding focusing abilities, which make them able to exert forces on micro-sized particles. The magnitude of such forces (pN) is strong enough to trap their targets. However, the most conventional OT setups are based on complex configurations, being associated with focusing difficulties with biologic samples. Optical Fiber Tweezers (OFTs), which consist in optical fibers with a lens in one of its extremities are valuable alternatives to Conventional Optical Tweezers (COTs). OFTs are flexible, simpler, low-cost and easy to handle. However, its trapping performance when manipulating biological and complex structures remains poorly characterized. In this study, we experimentally characterized the optical trapping of a biological cell found within a culture of rodent glial neuronal cells, using a polymeric lens fabricated through a photo-polymerization method on the top of a fiber. Its trapping performance was compared with two synthetic microspheres (PMMA, polystyrene) and two simple cells (a yeast and a Drosophila Melanogaster cell). Moreover, the experimental results were also compared with theoretical calculations made using a numerical model based on the Finite Differences Time Domain. It was found that, although the mammalian neuronal cell had larger dimensions, the magnitude of forces exerted on it was the lowest among all particles. Our results allowed us to quantify, for the first time, the complexity degree of manipulating such "demanding" cells in comparison with known targets. Thus, they can provide valuable insights about the influence of particle parameters such as size, refractive index, homogeneity degree and nature (biologic, synthetic). Furthermore, the theoretical results matched the experimental ones which validates the proposed model.

2018

Fabrication of Multimode-Single Mode Polymer Fiber Tweezers for Single Cell Trapping and Identification with Improved Performance

Autores
Rodrigues, SM; Paiva, JS; Ribeiro, RSR; Soppera, O; Cunha, JPS; Jorge, PAS;

Publicação
SENSORS

Abstract
Optical fiber tweezers have been gaining prominence in several applications in Biology and Medicine. Due to their outstanding focusing abilities, they are able to trap and manipulate microparticles, including cells, needing any physical contact and with a low degree of invasiveness to the trapped cell. Recently, we proposed a fiber tweezer configuration based on a polymeric micro-lens on the top of a single mode fiber, obtained by a self-guided photopolymerization process. This configuration is able to both trap and identify the target through the analysis of short-term portions of the back-scattered signal. In this paper, we propose a variant of this fabrication method, capable of producing more robust fiber tips, which produce stronger trapping effects on targets by as much as two to ten fold. These novel lenses maintain the capability of distinguish the different classes of trapped particles based on the back-scattered signal. This novel fabrication method consists in the introduction of a multi mode fiber section on the tip of a single mode (SM) fiber. A detailed description of how relevant fabrication parameters such as the length of the multi mode section and the photopolymerization laser power can be tuned for different purposes (e.g., microparticles trapping only, simultaneous trapping and sensing) is also provided, based on both experimental and theoretical evidences.

2018

Towards a Single Parameter Sensing for Bacteria Sorting through Optical Fiber Trapping and Back-Scattered Signal Analysis

Autores
Paiva, JS; Ribeiro, RSR; Jorge, PAS; Rosa, CC; Cunha, JPS;

Publicação
26th International Conference on Optical Fiber Sensors

Abstract

2018

Improved fabrication of polymeric optical fiber tweezers for single cell detection

Autores
Rodrigues, SM; Paiva, JS; Ribeiro, RSR; Soppera, O; Jorge, PAS;

Publicação
Optics InfoBase Conference Papers

Abstract
A new fabrication method of polymeric optical fiber tweezers with a multi-mode tip is presented. Preliminary results show higher robustness, improved ability for 2D trapping and differentiation of particles based on back-scattering analysis. © OSA 2018 © 2018 The Author(s)

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