Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence

Rafal Luchowski, Tanya Shtoyko, Elisa Apicella, Pabak Sarkar, Irina Akopova, Sangram Limbaji Raut, Rafal Fudala, Julian Borejdo, Zygmunt Gryczynski, Ignacy Gryczynski

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

This paper describes a method to significantly enhance single-molecule fluorescence detection in confocal microscopy, demonstrating that fractallike silver structures significantly improve dye stability and brightness. The experiments compared two immunoassay models based on the deposition of rabbit IgG on silver structures. The experiments were performed with the fluorophore-labeled protein at low picomolar concentrations. Well-separated bright spots were still easily distinguishable. Under standardized conditions we observed increased photostability and brightness enhancement for the dyes that were immobilized on the surface using a primary antibody. In contrast, when the unlabeled primary antibody was immobilized on the surface and the labeled secondary antibody was placed at a larger distance, we observed only a modest enhancement of fluorescence. Furthermore, based on backscattered reflected light images, it was proven that the observed fluorescence enhancements originate from the areas with deposited silver nanostructures. Fractal-like substrates are relatively easy to prepare. We believe that with their superior performance, they should find wide applications in single-molecule studies in which a longer observation time is required.

Original languageEnglish
Pages (from-to)174-180
Number of pages7
JournalApplied Spectroscopy
Volume65
Issue number2
DOIs
StatePublished - 1 Feb 2011

Fingerprint

antibodies
Silver
Antibodies
Fractals
Luminance
fractals
brightness
Fluorescence
silver
fluorescence
Molecules
augmentation
Coloring Agents
Dyes
dyes
molecules
immunoassay
Fluorophores
Confocal microscopy
rabbits

Keywords

  • FLIM
  • Fluorescence lifetime imaging microscopy
  • Fractal-like structures
  • Metalenhanced fluorescence
  • Single-molecule detection

Cite this

Luchowski, Rafal ; Shtoyko, Tanya ; Apicella, Elisa ; Sarkar, Pabak ; Akopova, Irina ; Raut, Sangram Limbaji ; Fudala, Rafal ; Borejdo, Julian ; Gryczynski, Zygmunt ; Gryczynski, Ignacy. / Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence. In: Applied Spectroscopy. 2011 ; Vol. 65, No. 2. pp. 174-180.
@article{526f4638ebb94053b4e5a5f32c635db1,
title = "Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence",
abstract = "This paper describes a method to significantly enhance single-molecule fluorescence detection in confocal microscopy, demonstrating that fractallike silver structures significantly improve dye stability and brightness. The experiments compared two immunoassay models based on the deposition of rabbit IgG on silver structures. The experiments were performed with the fluorophore-labeled protein at low picomolar concentrations. Well-separated bright spots were still easily distinguishable. Under standardized conditions we observed increased photostability and brightness enhancement for the dyes that were immobilized on the surface using a primary antibody. In contrast, when the unlabeled primary antibody was immobilized on the surface and the labeled secondary antibody was placed at a larger distance, we observed only a modest enhancement of fluorescence. Furthermore, based on backscattered reflected light images, it was proven that the observed fluorescence enhancements originate from the areas with deposited silver nanostructures. Fractal-like substrates are relatively easy to prepare. We believe that with their superior performance, they should find wide applications in single-molecule studies in which a longer observation time is required.",
keywords = "FLIM, Fluorescence lifetime imaging microscopy, Fractal-like structures, Metalenhanced fluorescence, Single-molecule detection",
author = "Rafal Luchowski and Tanya Shtoyko and Elisa Apicella and Pabak Sarkar and Irina Akopova and Raut, {Sangram Limbaji} and Rafal Fudala and Julian Borejdo and Zygmunt Gryczynski and Ignacy Gryczynski",
year = "2011",
month = "2",
day = "1",
doi = "10.1366/10-06112",
language = "English",
volume = "65",
pages = "174--180",
journal = "Applied Spectroscopy",
issn = "0003-7028",
publisher = "SAGE Publications Inc.",
number = "2",

}

Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence. / Luchowski, Rafal; Shtoyko, Tanya; Apicella, Elisa; Sarkar, Pabak; Akopova, Irina; Raut, Sangram Limbaji; Fudala, Rafal; Borejdo, Julian; Gryczynski, Zygmunt; Gryczynski, Ignacy.

In: Applied Spectroscopy, Vol. 65, No. 2, 01.02.2011, p. 174-180.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence

AU - Luchowski, Rafal

AU - Shtoyko, Tanya

AU - Apicella, Elisa

AU - Sarkar, Pabak

AU - Akopova, Irina

AU - Raut, Sangram Limbaji

AU - Fudala, Rafal

AU - Borejdo, Julian

AU - Gryczynski, Zygmunt

AU - Gryczynski, Ignacy

PY - 2011/2/1

Y1 - 2011/2/1

N2 - This paper describes a method to significantly enhance single-molecule fluorescence detection in confocal microscopy, demonstrating that fractallike silver structures significantly improve dye stability and brightness. The experiments compared two immunoassay models based on the deposition of rabbit IgG on silver structures. The experiments were performed with the fluorophore-labeled protein at low picomolar concentrations. Well-separated bright spots were still easily distinguishable. Under standardized conditions we observed increased photostability and brightness enhancement for the dyes that were immobilized on the surface using a primary antibody. In contrast, when the unlabeled primary antibody was immobilized on the surface and the labeled secondary antibody was placed at a larger distance, we observed only a modest enhancement of fluorescence. Furthermore, based on backscattered reflected light images, it was proven that the observed fluorescence enhancements originate from the areas with deposited silver nanostructures. Fractal-like substrates are relatively easy to prepare. We believe that with their superior performance, they should find wide applications in single-molecule studies in which a longer observation time is required.

AB - This paper describes a method to significantly enhance single-molecule fluorescence detection in confocal microscopy, demonstrating that fractallike silver structures significantly improve dye stability and brightness. The experiments compared two immunoassay models based on the deposition of rabbit IgG on silver structures. The experiments were performed with the fluorophore-labeled protein at low picomolar concentrations. Well-separated bright spots were still easily distinguishable. Under standardized conditions we observed increased photostability and brightness enhancement for the dyes that were immobilized on the surface using a primary antibody. In contrast, when the unlabeled primary antibody was immobilized on the surface and the labeled secondary antibody was placed at a larger distance, we observed only a modest enhancement of fluorescence. Furthermore, based on backscattered reflected light images, it was proven that the observed fluorescence enhancements originate from the areas with deposited silver nanostructures. Fractal-like substrates are relatively easy to prepare. We believe that with their superior performance, they should find wide applications in single-molecule studies in which a longer observation time is required.

KW - FLIM

KW - Fluorescence lifetime imaging microscopy

KW - Fractal-like structures

KW - Metalenhanced fluorescence

KW - Single-molecule detection

UR - http://www.scopus.com/inward/record.url?scp=79951907038&partnerID=8YFLogxK

U2 - 10.1366/10-06112

DO - 10.1366/10-06112

M3 - Article

AN - SCOPUS:79951907038

VL - 65

SP - 174

EP - 180

JO - Applied Spectroscopy

JF - Applied Spectroscopy

SN - 0003-7028

IS - 2

ER -

Luchowski R, Shtoyko T, Apicella E, Sarkar P, Akopova I, Raut SL et al. Fractal-like silver aggregates enhance the brightness and stability of single-molecule fluorescence. Applied Spectroscopy. 2011 Feb 1;65(2):174-180. https://doi.org/10.1366/10-06112