New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS)

Zygmunt Gryczynski, Ignacy Gryczynski, E. G. Matveeva, N. Calander, R. Grygorczyk, I. Akopova, S. Bharill, P. Muthu, S. Klidgar, Julian Borejdo

Research output: Chapter in Book/Report/Conference proceedingConference contributionResearchpeer-review

4 Citations (Scopus)

Abstract

We report new approach to Fluorescence Correlation Spectroscopy (FCS) and Single Molecule Detection (SMD) based on Surface Plasmon-Coupled Emission (SPCE) technology. The use of SPCE offers significant reduction of fluorescence volume (detection volume) reduction decreasing background contribution. Fluorophore interaction with surface plasmons increases the rate of photon detection and makes fluorescence very sensitive to change in a position of emitting molecule. The effective thickness of the fluorescence volume in SPCE experiments depends on two factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The excitation with the laser beam at Surface Plasmon Resonance (SPR) angle (Kretschmann configuration) through the high numerical aperture objective makes observation volume very shallow below 100 nm. The layer thickness is further reduced by the metal quenching of excited fluorophores immediately close to the interface (∼10 nm). The fluorescence light is emitted through the metal film only at the SPCE angle. Any fluorescence occurring at the distances greater than the coupling distance is effectively reflected (∼92%) by the metal film and not transmitted to the objective. The thickness of the detected volume can be 20-50 nm, depending on the prism dielectric constants and orientation of the excited dipoles. In addition the signal is very sensitive to the change in fluorophore position and orientation. Such strong dependence of the coupling to the surface plasmons on the orientation of excited dipoles opens new possibilities to study conformational changes of macromolecules in real time.

Original languageEnglish
Title of host publicationUltrasensitive and Single-Molecule Detection Technologies II
Volume6444
DOIs
StatePublished - 30 Apr 2007
EventUltrasensitive and Single-Molecule Detection Technologies II - San Jose, CA, United States
Duration: 20 Jan 200723 Jan 2007

Other

OtherUltrasensitive and Single-Molecule Detection Technologies II
CountryUnited States
CitySan Jose, CA
Period20/01/0723/01/07

Fingerprint

Plasmons
Fluorescence Spectrometry
plasmons
Fluorescence
Spectroscopy
Fluorophores
fluorescence
Molecules
spectroscopy
Metals
molecules
metal films
Surface Plasmon Resonance
Photons
dipoles
Lasers
evanescent waves
wave excitation
Surface plasmon resonance
numerical aperture

Keywords

  • Fluorescence
  • Fluorescence correlation spectroscopy
  • Macromolecular conformational changes
  • Minimized detection volume
  • Surface plasmon-coupled emission

Cite this

Gryczynski, Z., Gryczynski, I., Matveeva, E. G., Calander, N., Grygorczyk, R., Akopova, I., ... Borejdo, J. (2007). New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS). In Ultrasensitive and Single-Molecule Detection Technologies II (Vol. 6444). [64440G] https://doi.org/10.1117/12.715268
Gryczynski, Zygmunt ; Gryczynski, Ignacy ; Matveeva, E. G. ; Calander, N. ; Grygorczyk, R. ; Akopova, I. ; Bharill, S. ; Muthu, P. ; Klidgar, S. ; Borejdo, Julian. / New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS). Ultrasensitive and Single-Molecule Detection Technologies II. Vol. 6444 2007.
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abstract = "We report new approach to Fluorescence Correlation Spectroscopy (FCS) and Single Molecule Detection (SMD) based on Surface Plasmon-Coupled Emission (SPCE) technology. The use of SPCE offers significant reduction of fluorescence volume (detection volume) reduction decreasing background contribution. Fluorophore interaction with surface plasmons increases the rate of photon detection and makes fluorescence very sensitive to change in a position of emitting molecule. The effective thickness of the fluorescence volume in SPCE experiments depends on two factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The excitation with the laser beam at Surface Plasmon Resonance (SPR) angle (Kretschmann configuration) through the high numerical aperture objective makes observation volume very shallow below 100 nm. The layer thickness is further reduced by the metal quenching of excited fluorophores immediately close to the interface (∼10 nm). The fluorescence light is emitted through the metal film only at the SPCE angle. Any fluorescence occurring at the distances greater than the coupling distance is effectively reflected (∼92{\%}) by the metal film and not transmitted to the objective. The thickness of the detected volume can be 20-50 nm, depending on the prism dielectric constants and orientation of the excited dipoles. In addition the signal is very sensitive to the change in fluorophore position and orientation. Such strong dependence of the coupling to the surface plasmons on the orientation of excited dipoles opens new possibilities to study conformational changes of macromolecules in real time.",
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author = "Zygmunt Gryczynski and Ignacy Gryczynski and Matveeva, {E. G.} and N. Calander and R. Grygorczyk and I. Akopova and S. Bharill and P. Muthu and S. Klidgar and Julian Borejdo",
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Gryczynski, Z, Gryczynski, I, Matveeva, EG, Calander, N, Grygorczyk, R, Akopova, I, Bharill, S, Muthu, P, Klidgar, S & Borejdo, J 2007, New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS). in Ultrasensitive and Single-Molecule Detection Technologies II. vol. 6444, 64440G, Ultrasensitive and Single-Molecule Detection Technologies II, San Jose, CA, United States, 20/01/07. https://doi.org/10.1117/12.715268

New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS). / Gryczynski, Zygmunt; Gryczynski, Ignacy; Matveeva, E. G.; Calander, N.; Grygorczyk, R.; Akopova, I.; Bharill, S.; Muthu, P.; Klidgar, S.; Borejdo, Julian.

Ultrasensitive and Single-Molecule Detection Technologies II. Vol. 6444 2007. 64440G.

Research output: Chapter in Book/Report/Conference proceedingConference contributionResearchpeer-review

TY - GEN

T1 - New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS)

AU - Gryczynski, Zygmunt

AU - Gryczynski, Ignacy

AU - Matveeva, E. G.

AU - Calander, N.

AU - Grygorczyk, R.

AU - Akopova, I.

AU - Bharill, S.

AU - Muthu, P.

AU - Klidgar, S.

AU - Borejdo, Julian

PY - 2007/4/30

Y1 - 2007/4/30

N2 - We report new approach to Fluorescence Correlation Spectroscopy (FCS) and Single Molecule Detection (SMD) based on Surface Plasmon-Coupled Emission (SPCE) technology. The use of SPCE offers significant reduction of fluorescence volume (detection volume) reduction decreasing background contribution. Fluorophore interaction with surface plasmons increases the rate of photon detection and makes fluorescence very sensitive to change in a position of emitting molecule. The effective thickness of the fluorescence volume in SPCE experiments depends on two factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The excitation with the laser beam at Surface Plasmon Resonance (SPR) angle (Kretschmann configuration) through the high numerical aperture objective makes observation volume very shallow below 100 nm. The layer thickness is further reduced by the metal quenching of excited fluorophores immediately close to the interface (∼10 nm). The fluorescence light is emitted through the metal film only at the SPCE angle. Any fluorescence occurring at the distances greater than the coupling distance is effectively reflected (∼92%) by the metal film and not transmitted to the objective. The thickness of the detected volume can be 20-50 nm, depending on the prism dielectric constants and orientation of the excited dipoles. In addition the signal is very sensitive to the change in fluorophore position and orientation. Such strong dependence of the coupling to the surface plasmons on the orientation of excited dipoles opens new possibilities to study conformational changes of macromolecules in real time.

AB - We report new approach to Fluorescence Correlation Spectroscopy (FCS) and Single Molecule Detection (SMD) based on Surface Plasmon-Coupled Emission (SPCE) technology. The use of SPCE offers significant reduction of fluorescence volume (detection volume) reduction decreasing background contribution. Fluorophore interaction with surface plasmons increases the rate of photon detection and makes fluorescence very sensitive to change in a position of emitting molecule. The effective thickness of the fluorescence volume in SPCE experiments depends on two factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The excitation with the laser beam at Surface Plasmon Resonance (SPR) angle (Kretschmann configuration) through the high numerical aperture objective makes observation volume very shallow below 100 nm. The layer thickness is further reduced by the metal quenching of excited fluorophores immediately close to the interface (∼10 nm). The fluorescence light is emitted through the metal film only at the SPCE angle. Any fluorescence occurring at the distances greater than the coupling distance is effectively reflected (∼92%) by the metal film and not transmitted to the objective. The thickness of the detected volume can be 20-50 nm, depending on the prism dielectric constants and orientation of the excited dipoles. In addition the signal is very sensitive to the change in fluorophore position and orientation. Such strong dependence of the coupling to the surface plasmons on the orientation of excited dipoles opens new possibilities to study conformational changes of macromolecules in real time.

KW - Fluorescence

KW - Fluorescence correlation spectroscopy

KW - Macromolecular conformational changes

KW - Minimized detection volume

KW - Surface plasmon-coupled emission

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U2 - 10.1117/12.715268

DO - 10.1117/12.715268

M3 - Conference contribution

SN - 0819465577

SN - 9780819465573

VL - 6444

BT - Ultrasensitive and Single-Molecule Detection Technologies II

ER -

Gryczynski Z, Gryczynski I, Matveeva EG, Calander N, Grygorczyk R, Akopova I et al. New surface plasmons approach to Single Molecule Detection (SMD) and Fluorescence Correlation Spectroscopy (FCS). In Ultrasensitive and Single-Molecule Detection Technologies II. Vol. 6444. 2007. 64440G https://doi.org/10.1117/12.715268