Single Nucleotide Polymorphism Detection Using Gold Nanoprobes and Bio-Microfluidic Platform With Embedded Microlenses

Iwona Bernacka-Wojcik, Hugo Aguas, Fabio Ferreira Carlos, Paulo Lopes, Pawel Jerzy Wojcik, Mafalda Nascimento Costa, Bruno Veigas, Rui Igreja, Elvira Fortunato, Pedro Baptista, Rodrigo Martins

Research output: Contribution to journalArticle

  • 5 Citations

Abstract

The use of microfluidics platforms combined with the optimal optical properties of gold nanopartides has found plenty of application in molecular biosensing. This paper describes a biotnicrofluidic platform coupled to a non-cross-linking colorimetric gold nanoprobe assay to detect a single nucleotide polymorphism associated with increased risk of obesity fat-mass and obesity-associated (FTO) rs9939609 (Carlos et al., 2014). The system enabled significant discrimination between positive and negative assays using a target DNA concentration of 5 ng/mu l below the limit of detection of the conventionally used microplate reader (i.e., 15 ng/mu l) with 10 times lower solution volume (i.e., 3 mu l.). A set of optimization of our previously reported bio-microfluidic platform (Bemacka-Wojcik et al., 2013) resulted in a 160% improvement of colorimetric analysis results. Incorporation of planar microlenses increased 6 times signal-to-loss ratio reaching the output optical fiber improving by 34% the colorimetric analysis of gold nanopartides, while the implementation of an optoelectronic acquisition system yielded increased accuracy and reduced noise. The microfluidic chip was also integrated with a miniature fiber spectrometer to analyze the assays' cobrimetric changes and also the LEDs transmission spectra when illuminating through various solutions. Furthermore, by coupling an optical micmscope to a digital camera with a long exposure time (30s), we could visualise the different scatter intensities of gold nanoparticles within channels following salt addition. These intensities correlate well to the expected difference in aggregation between FTO positive (none to small aggregates) and negative samples (large aggregates). (C) 2015 Wiley Periodicals, Inc.

LanguageEnglish
Pages1210-1219
Number of pages10
JournalBiotechnology and Bioengineering
Volume112
Issue number6
DOIs
StatePublished - 10 Mar 2015

Keywords

  • microfluidics
  • DNA
  • gold nanoparticles
  • fiberoptics
  • single nucleotide polymorphism
  • MYCOBACTERIUM-TUBERCULOSIS
  • CHIP
  • AMPLIFICATION
  • NANOPARTICLES
  • DIAGNOSTICS
  • DEVICES
  • BIOCHIP
  • SILVER
  • PCR

Cite this

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title = "Single Nucleotide Polymorphism Detection Using Gold Nanoprobes and Bio-Microfluidic Platform With Embedded Microlenses",
abstract = "The use of microfluidics platforms combined with the optimal optical properties of gold nanopartides has found plenty of application in molecular biosensing. This paper describes a biotnicrofluidic platform coupled to a non-cross-linking colorimetric gold nanoprobe assay to detect a single nucleotide polymorphism associated with increased risk of obesity fat-mass and obesity-associated (FTO) rs9939609 (Carlos et al., 2014). The system enabled significant discrimination between positive and negative assays using a target DNA concentration of 5 ng/mu l below the limit of detection of the conventionally used microplate reader (i.e., 15 ng/mu l) with 10 times lower solution volume (i.e., 3 mu l.). A set of optimization of our previously reported bio-microfluidic platform (Bemacka-Wojcik et al., 2013) resulted in a 160{\%} improvement of colorimetric analysis results. Incorporation of planar microlenses increased 6 times signal-to-loss ratio reaching the output optical fiber improving by 34{\%} the colorimetric analysis of gold nanopartides, while the implementation of an optoelectronic acquisition system yielded increased accuracy and reduced noise. The microfluidic chip was also integrated with a miniature fiber spectrometer to analyze the assays' cobrimetric changes and also the LEDs transmission spectra when illuminating through various solutions. Furthermore, by coupling an optical micmscope to a digital camera with a long exposure time (30s), we could visualise the different scatter intensities of gold nanoparticles within channels following salt addition. These intensities correlate well to the expected difference in aggregation between FTO positive (none to small aggregates) and negative samples (large aggregates). (C) 2015 Wiley Periodicals, Inc.",
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author = "Iwona Bernacka-Wojcik and Hugo Aguas and Carlos, {Fabio Ferreira} and Paulo Lopes and Wojcik, {Pawel Jerzy} and Costa, {Mafalda Nascimento} and Bruno Veigas and Rui Igreja and Elvira Fortunato and Pedro Baptista and Rodrigo Martins",
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pages = "1210--1219",
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Single Nucleotide Polymorphism Detection Using Gold Nanoprobes and Bio-Microfluidic Platform With Embedded Microlenses. / Bernacka-Wojcik, Iwona; Aguas, Hugo; Carlos, Fabio Ferreira; Lopes, Paulo; Wojcik, Pawel Jerzy; Costa, Mafalda Nascimento; Veigas, Bruno; Igreja, Rui; Fortunato, Elvira; Baptista, Pedro; Martins, Rodrigo.

In: Biotechnology and Bioengineering, Vol. 112, No. 6, 10.03.2015, p. 1210-1219.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Single Nucleotide Polymorphism Detection Using Gold Nanoprobes and Bio-Microfluidic Platform With Embedded Microlenses

AU - Bernacka-Wojcik,Iwona

AU - Aguas,Hugo

AU - Carlos,Fabio Ferreira

AU - Lopes,Paulo

AU - Wojcik,Pawel Jerzy

AU - Costa,Mafalda Nascimento

AU - Veigas,Bruno

AU - Igreja,Rui

AU - Fortunato,Elvira

AU - Baptista,Pedro

AU - Martins,Rodrigo

PY - 2015/3/10

Y1 - 2015/3/10

N2 - The use of microfluidics platforms combined with the optimal optical properties of gold nanopartides has found plenty of application in molecular biosensing. This paper describes a biotnicrofluidic platform coupled to a non-cross-linking colorimetric gold nanoprobe assay to detect a single nucleotide polymorphism associated with increased risk of obesity fat-mass and obesity-associated (FTO) rs9939609 (Carlos et al., 2014). The system enabled significant discrimination between positive and negative assays using a target DNA concentration of 5 ng/mu l below the limit of detection of the conventionally used microplate reader (i.e., 15 ng/mu l) with 10 times lower solution volume (i.e., 3 mu l.). A set of optimization of our previously reported bio-microfluidic platform (Bemacka-Wojcik et al., 2013) resulted in a 160% improvement of colorimetric analysis results. Incorporation of planar microlenses increased 6 times signal-to-loss ratio reaching the output optical fiber improving by 34% the colorimetric analysis of gold nanopartides, while the implementation of an optoelectronic acquisition system yielded increased accuracy and reduced noise. The microfluidic chip was also integrated with a miniature fiber spectrometer to analyze the assays' cobrimetric changes and also the LEDs transmission spectra when illuminating through various solutions. Furthermore, by coupling an optical micmscope to a digital camera with a long exposure time (30s), we could visualise the different scatter intensities of gold nanoparticles within channels following salt addition. These intensities correlate well to the expected difference in aggregation between FTO positive (none to small aggregates) and negative samples (large aggregates). (C) 2015 Wiley Periodicals, Inc.

AB - The use of microfluidics platforms combined with the optimal optical properties of gold nanopartides has found plenty of application in molecular biosensing. This paper describes a biotnicrofluidic platform coupled to a non-cross-linking colorimetric gold nanoprobe assay to detect a single nucleotide polymorphism associated with increased risk of obesity fat-mass and obesity-associated (FTO) rs9939609 (Carlos et al., 2014). The system enabled significant discrimination between positive and negative assays using a target DNA concentration of 5 ng/mu l below the limit of detection of the conventionally used microplate reader (i.e., 15 ng/mu l) with 10 times lower solution volume (i.e., 3 mu l.). A set of optimization of our previously reported bio-microfluidic platform (Bemacka-Wojcik et al., 2013) resulted in a 160% improvement of colorimetric analysis results. Incorporation of planar microlenses increased 6 times signal-to-loss ratio reaching the output optical fiber improving by 34% the colorimetric analysis of gold nanopartides, while the implementation of an optoelectronic acquisition system yielded increased accuracy and reduced noise. The microfluidic chip was also integrated with a miniature fiber spectrometer to analyze the assays' cobrimetric changes and also the LEDs transmission spectra when illuminating through various solutions. Furthermore, by coupling an optical micmscope to a digital camera with a long exposure time (30s), we could visualise the different scatter intensities of gold nanoparticles within channels following salt addition. These intensities correlate well to the expected difference in aggregation between FTO positive (none to small aggregates) and negative samples (large aggregates). (C) 2015 Wiley Periodicals, Inc.

KW - microfluidics

KW - DNA

KW - gold nanoparticles

KW - fiberoptics

KW - single nucleotide polymorphism

KW - MYCOBACTERIUM-TUBERCULOSIS

KW - CHIP

KW - AMPLIFICATION

KW - NANOPARTICLES

KW - DIAGNOSTICS

KW - DEVICES

KW - BIOCHIP

KW - SILVER

KW - PCR

U2 - 10.1002/bit.25542

DO - 10.1002/bit.25542

M3 - Article

VL - 112

SP - 1210

EP - 1219

JO - Biotechnology and Bioengineering

T2 - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 6

ER -