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Immunogold Electron Microscopy01:20

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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis
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Surface enhanced fluorescence immuno-biosensor based on gold nanorods.

Linus Pauling F Peixoto1, Jacqueline F L Santos2, Gustavo F S Andrade1

  • 1Laboratório de Nanoestruturas Plasmônicas, Núcleo de Espectroscopia e Estrutura Molecular, Centro de Estudos em Materiais, Departamento de Química, Universidade Federal de Juiz de Fora, Juiz De Fora, MG, Brazil.

Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
|September 4, 2022
PubMed
Summary

This study presents a novel gold nanorod (AuNR)-based biosensor for detecting albumin antibodies. The localized surface plasmon resonance (LSPR) property enhances fluorescence detection, enabling rapid and specific immunoassays.

Keywords:
BiosensorsEnhanced spectroscopiesMolecular recognitionNanoparticlesPlasmonicsSEF

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Area of Science:

  • Nanotechnology
  • Biomedical Engineering
  • Analytical Chemistry

Background:

  • Gold nanoparticles (AuNPs) offer unique nanoscale properties and biocompatibility for biosensing applications.
  • Localized surface plasmon resonance (LSPR) enhances electromagnetic fields near metallic surfaces, enabling techniques like surface-enhanced fluorescence (SEF).

Purpose of the Study:

  • To construct a simple, fast immuno-biosensor using gold nanorods (AuNRs) and SEF for detecting albumin antibodies (anti-BSA).
  • To utilize the antigen-antibody (anti-BSA/BSA) interaction as a biorecognition model for the immunoassay.

Main Methods:

  • AuNRs were prepared in suspension (S-AuNRs) and immobilized on glass slides (AuNRs-chip).
  • An extrinsic SEF method monitored the signal of a reporter dye (IR-820) to detect anti-BSA.
  • SEF mapping and a digital protocol were employed for signal analysis and variability reduction.

Main Results:

  • Analyte detection was confirmed by SEF mapping, showing a threefold increase in signal intensity with anti-BSA compared to its absence.
  • The digital protocol indicated approximately a twofold increase in positive events when anti-BSA was present versus absent.
  • The AuNRs-based SEF immuno-biosensor demonstrated efficient and specific biorecognition.

Conclusions:

  • The developed AuNRs-based SEF immuno-biosensor provides an efficient and simple platform for anti-BSA detection.
  • This approach offers specific biorecognition and may serve as a valuable spectroscopy platform for future immuno-biosensing applications.