Multi-wavelength immunoassays using surface plasmon-coupled emission

doi:10.1016/j.bbrc.2003.12.010

Biochemical and Biophysical Research Communications

Volume 313, Issue 3, 16 January 2004, Pages 721-726

https://doi.org/10.1016/j.bbrc.2003.12.010 Get rights and content

Abstract

We describe a new method for multi-wavelength immunoassays using surface plasmon-coupled emission (SPCE). This phenomenon is coupling of excited fluorophores with a nearby thin metal film, in our case silver, resulting in strongly directional emission into the underlying glass substrate. The angle at which the radiation propagate through the prism depends on the surface plasmon angle for the relevant wavelength. These angles depend on emission wavelength, allowing measurement of multiple analytes using multiple emission wavelengths. We demonstrated this possibility using antibodies labeled with either Rhodamine Red-X or AlexaFluor 647. These antibodies were directed against an antigen protein bound to the silver surface. The emission from each labeled antibody occurred at a different angle on the glass prism, allowing independent measurement of surface binding of each antibody. This method of SPCE immunoassays can be readily extended to 4 or more wavelengths.

Section snippets

Materials and methods

Reagents. Rabbit IgG (11.2 mg/mL) was from Sigma. Rhodamine Red-X-antiRabbit IgG (produced in goat, 2 mg/mL, dye/protein=3.8 mol/mol) and AlexaFluor647-antiRabbit IgG (produced in goat, 2 mg/mL, dye/protein=4.5 mol/mol) conjugates were from Molecular Probes. Buffer components and salts (such as bovine serum albumin, glucose, and sucrose) were from Sigma–Aldrich.

Coating slides with IgG. Standard glass microscope slides (3 × 1 in., 1 mm thick; Corning) were vapor deposited with a continuous 50 nm thick

Theory

The phenomenon of SPCE is closely related to surface plasmon resonance (SPR). The theory of surface plasmons has been described in details [16], [17] as have its use to measure bioaffinity reactions [18], [19]. Surface plasmons are excited when a metal surface is illuminated under specific conditions which allow wavevector matching at the sample metal interface. Surface plasmons cannot be excited from air by incident light. SPR occurs when light is incident on a metal through a higher

Results

The experimental configuration used for two-wavelength SPCE is shown in Scheme 2. The protein-coated silver surface is illuminated at the surface plasmon angle through the glass prism, which is called the Kretschmann (KR) configuration. The free-space emission is observed normal to the sample surface, on the side distal from the prism, using a filter and a fiber optic bundle. SPCE is observed on the prism side of the sample, at two different angles through appropriate long-pass filters for each

Discussion

We expect angle and wavelength-dependent SPCE to have numerous applications in sensing. The silver film serves multiple purposes. It amplifies the incident light [18], efficiently collects the emission, and provides separation of the wavelengths [12], [13]. Detection could be accomplished with imaging or point detectors, to provide a simple yet sensitive device. The number of analytes can be increased by using fluorophores with emission wavelengths ranging from 450 to 800 nm. Still more analytes

Acknowledgements

This work was supported by the National Institute of Biomedical Imaging and Bioengineering, EB-00682 and EB-00981, the National Center for Research Resource, RR-08119 and Philip Morris USA, Inc.

References (33)

T. Vo-Dinh et al.
Immunosensors: principles and applications
Immunomethods
(1993)
W.B. Dandliker et al.
Fluorescence polarization in immunochemistry
Immunochemistry
(1970)
L.E. Morrison
Time-resolved detection of energy transfer: theory and application to immunoassays
Anal. Biochem.
(1988)
E.F. Ullman et al.
Fluorescent excitation transfer immunoassay: a general method for determination of antigens
J. Biol. Chem.
(1976)
E.P. Diamandis
Immunoassays with time-resolved fluorescence spectroscopy: principles and applications
Clin. Biochem.
(1988)
J.R. Lakowicz
Radiative decay engineering. Biophysical and biomedical applications
Anal. Biochem.
(2001)
J.R. Lakowicz et al.
Radiative decay engineering. 2. Effects of silver island films on fluorescence intensity, lifetimes and resonance energy transfer
Anal. Biochem.
(2002)
Z. Salamon et al.
Surface plasmon resonance spectroscopy as a tool for investigating the biochemical and biophysical properties of membrane protein systems. I: theoretical principles
Biochim. Biophys. Acta
(1997)
J. Melendez et al.
A commercial solution for surface plasmon sensing
Sensors Actuators B
(1996)
T. Liebermann et al.
Surface-plasmon field-enhanced fluorescence spectroscopy
Colloids Surf.
(2000)

S.C. Kitson et al.

Photoluminescence from dye molecules on silver gratings

Opt. Commun.

(1996)

G.L. Duveneck et al.

Two-photon fluorescence excitation of macroscopic areas on planar waveguides

Biosensors Bioelectron.

(2003)

I.A. Hemmila

Applications of Fluorescence in Immunoassays

(1992)

M. Fiore et al.

The abbott IMx^TM automated benchtop immunochemistry analyzer system

Clin. Chem.

(1988)

T. Lövgren et al.

Time-resolved fluoroimmunoassay, advantages and limitations

Cited by (70)

Design and numerical simulation of a sensitive plasmonic-based nanosensor utilizing MoS<inf>2</inf> monolayer and graphene
2023, Diamond and Related Materials
Citation Excerpt :
In recent years, surface plasmon resonance (SPR) biosensors have attracted much attention in designing optical sensors. The SPR-based structure is widely used for biosensing applications using a glass substrate coated with a thin metal layer [1–4]. This structure is proper for studying the interaction between immobilized biomolecules and a solution-phase analyte and presents interesting features such as high sensitivity, versatility, real-time, and label-free in comparison to past methods [5].
In this paper, a Kretschmann structure with a graphene-MoS₂ hybrid layer has been used to design a sensitive biosensor for the detection of biomolecules. Here, by using a silicon layer, the sensitivity of the nanosensor is increased. Such a result is obtained from strong light-matter interaction between graphene and semiconductor layer. As a result, by changing the active metal and adding an additional silicon layer at a wavelength of 670 nm to access higher sensitivity and lower FWHM for the effective biosensor eventually we reach a sensitivity equal to 192°/RIU. The designed structure is investigated by numerical analysis and using the FDTD method, we numerically extract the optical characteristics of the surface plasmon polariton sensor. The detection range for the proposed structure is obtained with a refractive index of 0.005. Also, the basic parameters of the sensor to achieve the best conditions have been calculated and evaluated. In addition, the overall size of the sensor is 1112 nm × 2000 nm.
Power dependent surface plasmon coupled emission studies of metal-dielectric-metal planar structure
2022, Materials Today: Proceedings
Citation Excerpt :
These methods give qualitative idea about the presence of molecules but lacks quantative features. On the other hand, the phenomenon of plasmon coupled emission [9,23-26] due to the interaction of fluorophores with nearby metallic films is of much interest due to fluorescence enhancement, control, steering and directionality. This is based on a reverse Kretschmann (RK) configuration [27-31].
Surface plasmon coupled emission (SPCE) was demonstrated in Metal-Dielectric-Metal Planar (MDMP) structure by coupling MDMP structure to a hemicylindrical prism by Reverse Kretchmann configuration. Here silver and Fluorescein dye dispersed in polyvinyl Alcohol (PVA) were used as metal and dielectric layers respectively. Due to the combined effect of (1) confinement of photon mode within MDMP structure and (2) excitation of plasmon modes by RK configuration, highly directional and enhanced emission was observed. Dependence of SPCE emissions on input excitation power were studied. Higher dielectric thickness (∼600 nm) shows a better emission efficiency as compared to lower dielectric thickness (∼140 nm). Our simulation results show that a higher field distribution within MDMP structure for dielectric thickness of 600 nm resulting in better surface plasmon emission coupling and hence in better emission efficiency.
Fluorescence molecular localization in submicronic depth through waveguide mode coupled emission
2020, Optics Communications
Citation Excerpt :
Leakage radiation microscopy (LRM) is a powerful tool for studying the interaction between near-field optics and micro-nano materials [1–6]. Various surface waves such as evanescent waves (EWs), surface plasmon polaritons (SPPs), Bloch surface waves (BSWs) can be characterized in the far field through coupled emission [7–15]. It has excellent applications in many fields such as label-free imaging, real-time measurements, optical tweezers, plasmon focusing, and so on [16–22].
The main challenge in exploiting surface wave for resolving the axial position of fluorophores is to maintain sufficient accuracy far from the interface. In this work, we explore the possibility of detecting the axial position of fluorescent molecules via the proportion of light emitted by independent modes. This technique is based on the modulation of fluorescent molecular radiation by waveguide slides and the coupled emission of corresponding modes. We compare the positioning accuracy in our method with previous work under the same noise conditions, and propose a multi-wavelength method for accurate positioning. The theoretical results provide a robust and simple implementation of axially localized detection for leakage radiation microscopy, which increase detection depth range and the real-time analysis ability.
Blood component detection based on miniaturized self-referenced hybrid Tamm-plasmon-polariton sensor
2018, Sensors and Actuators, B: Chemical
Citation Excerpt :
However, the conventional biochemical devices require significant laboratory space and are quite expensive by virtue of bulky optical sources as well as detection schemes. Surface plasmon resonance (SPR) based sensing schemes utilize the vulnerability of surface modes towards changes in ambient medium and finds wide application for numerous bio-sensing applications [5–7]. At present, SPR based sensing architectures have been employed in various detection schemes of biological as well as chemical entities such as DNA, RNA, pesticides etc.
A self-referenced sensor based on coupled hybrid mode Tamm-plasmon-polariton (TPP) is proposed for detection of different blood group components. The geometry which has been investigated theoretically, is comprised of blood components sandwiched between two ‘metal-distributed Bragg reflectors (DBRs)’ exhibiting two distinguishable absorption peaks within the photonic band-gap (PBG) of DBR. The two absorption maxima are essentially due to excitation of one symmetric (Mode-I) and one anti-symmetric mode (Mode-II) within the PBG. Mode-I remains invariant for a change of blood constituents whereas Mode-II exhibits strong dispersive properties by virtue of significant presence within the sensing region. This gives rise to a hybrid self-reference scheme where the longer wavelength arm (Mode-II) varies significantly providing proper signature of a particular blood component. A straight line fit to the resonance wavelength shift gives rise to a sensitivity of ≈200 nm/RIU with the proposed architecture. The polarization dependence of the spectral shift of hybrid-mode resonance wavelengths for oblique incidence is also investigated and analyzed theoretically. The sharp absorption maxima facilitate precise detection with an improved detection accuracy of a particular blood component. A brief analysis to optimize the sensor design for enhanced sensitivity, as well as detection accuracy, has also been discussed.
Plasmonic optical sensor for determination of refractive index of human skin tissues
2016, Sensors and Actuators, B: Chemical
Citation Excerpt :
Third, the resolution of measurement should be as small as possible so that even the negligibly small changes may be determined. There are a number of biological and biomedical parameters, where SPR-based biosensing has been proposed or utilized for their detection, such as feline calicivirus [3], antibodies [5], membrane proteins [7], immunoassays [8], human blood-groups [9], Avian influenza virus H5N1 [10], estrone [11], and hemoglobin concentration [12]. However, there are several other biological processes where SPR is yet to be explored.
A surface plasmon resonance (SPR) based optical sensor for the determination of refractive index of human skin tissue samples is proposed. Previous experimental results describing variation of refractive index of skin tissue samples with wavelength are considered for theoretical calculations. The angular interrogation method along with glass substrate and Au-Al bimetal layer is considered. The sensor's performance is closely analyzed in terms of well-defined performance indicators in order to achieve reliable and accurate sensing performance. The influence of operating wavelength on the performance of sensor scheme is investigated. Performance comparison for two different substrates (2S2G and SF10) is carried out. The proposed sensor has the potential to provide high sensitivity, precision, and resolution, thereby opening an easy and reliable window for dermatological applications.
Sensitivity enhancement of a grating-based surface plasmon-coupled emission (SPCE) biosensor chip using gold thickness
2014, Chemical Physics Letters
We describe a novel approach to enhance the sensitivity of a grating-based surface plasmon-coupled emission (SPCE) sensor by increasing the thickness of the metal film used in this system. The calculated optical properties of grating-based SPR spectra were significantly affected by both grating depth and by gold thickness. Higher angular sensitivity could be achieved at short wavelengths and under in situ measurement (analysis under aqueous condition). We confirmed the predicated enhancements of SPCE response using Alexa Fluor 647-labeled anti-mouse IgG immobilized on the SPCE sensor chips. Grating-coupled SPCE sensor chips can be used as a useful tool for high contents analysis of chemical and biomolecular interactions.

View all citing articles on Scopus

View full text

Multi-wavelength immunoassays using surface plasmon-coupled emission

Abstract

Section snippets

Materials and methods

Theory

Results

Discussion

Acknowledgements

Immunomethods

Immunochemistry

Anal. Biochem.

J. Biol. Chem.

Clin. Biochem.

Anal. Biochem.

Anal. Biochem.

Biochim. Biophys. Acta

Sensors Actuators B

Colloids Surf.

Opt. Commun.

Biosensors Bioelectron.

Applications of Fluorescence in Immunoassays

The abbott IMxTM automated benchtop immunochemistry analyzer system

Clin. Chem.

Time-resolved fluoroimmunoassay, advantages and limitations

The abbott IMx^TM automated benchtop immunochemistry analyzer system