Confocal, three-dimensional tracking of individual quantum dots in high-background environments

Nathan P. Wells; Guillaume A. Lessard; James H. Werner

doi:10.1021/ac8021899

Confocal, three-dimensional tracking of individual quantum dots in high-background environments

Nathan P. Wells
, Guillaume A. Lessard
, James H. Werner

Los Alamos National Laboratory

Research output: Contribution to journal › Article › peer-review

68 Scopus citations

Abstract

We demonstrate a custom confocal fluorescence-microscope that is capable of tracking individual quantum dots undergoing three-dimensional Brownian motion (diffusion coefficient ∼ 0.5 μm²/s) in environments with a signal-to-background ratio as low as 2:1, significantly worse than observed in a typical cellular environment. By utilizing a pulsed excitation source and time-correlated single photon counting, the time-resolved photon stream can be used to determine changes in the emission lifetime as a function of position and positively identify single quantum dots via photon-pair correlations. These results indicate that this microscope will be capable of following protein and RNA transport throughout the full three-dimensional volume of a live cell for durations up to 15 s.

Original language	English
Pages (from-to)	9830-9834
Number of pages	5
Journal	Analytical Chemistry
Volume	80
Issue number	24
DOIs	https://doi.org/10.1021/ac8021899
State	Published - Dec 15 2008

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title = "Confocal, three-dimensional tracking of individual quantum dots in high-background environments",

abstract = "We demonstrate a custom confocal fluorescence-microscope that is capable of tracking individual quantum dots undergoing three-dimensional Brownian motion (diffusion coefficient ∼ 0.5 μm2/s) in environments with a signal-to-background ratio as low as 2:1, significantly worse than observed in a typical cellular environment. By utilizing a pulsed excitation source and time-correlated single photon counting, the time-resolved photon stream can be used to determine changes in the emission lifetime as a function of position and positively identify single quantum dots via photon-pair correlations. These results indicate that this microscope will be capable of following protein and RNA transport throughout the full three-dimensional volume of a live cell for durations up to 15 s.",

author = "Wells, \{Nathan P.\} and Lessard, \{Guillaume A.\} and Werner, \{James H.\}",

year = "2008",

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day = "15",

doi = "10.1021/ac8021899",

language = "English",

volume = "80",

pages = "9830--9834",

journal = "Analytical Chemistry",

issn = "0003-2700",

publisher = "American Chemical Society",

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T1 - Confocal, three-dimensional tracking of individual quantum dots in high-background environments

AU - Wells, Nathan P.

AU - Lessard, Guillaume A.

AU - Werner, James H.

PY - 2008/12/15

Y1 - 2008/12/15

N2 - We demonstrate a custom confocal fluorescence-microscope that is capable of tracking individual quantum dots undergoing three-dimensional Brownian motion (diffusion coefficient ∼ 0.5 μm2/s) in environments with a signal-to-background ratio as low as 2:1, significantly worse than observed in a typical cellular environment. By utilizing a pulsed excitation source and time-correlated single photon counting, the time-resolved photon stream can be used to determine changes in the emission lifetime as a function of position and positively identify single quantum dots via photon-pair correlations. These results indicate that this microscope will be capable of following protein and RNA transport throughout the full three-dimensional volume of a live cell for durations up to 15 s.

AB - We demonstrate a custom confocal fluorescence-microscope that is capable of tracking individual quantum dots undergoing three-dimensional Brownian motion (diffusion coefficient ∼ 0.5 μm2/s) in environments with a signal-to-background ratio as low as 2:1, significantly worse than observed in a typical cellular environment. By utilizing a pulsed excitation source and time-correlated single photon counting, the time-resolved photon stream can be used to determine changes in the emission lifetime as a function of position and positively identify single quantum dots via photon-pair correlations. These results indicate that this microscope will be capable of following protein and RNA transport throughout the full three-dimensional volume of a live cell for durations up to 15 s.

UR - https://www.scopus.com/pages/publications/58149113044

U2 - 10.1021/ac8021899

DO - 10.1021/ac8021899

M3 - Article

SN - 0003-2700

VL - 80

SP - 9830

EP - 9834

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 24

ER -

Confocal, three-dimensional tracking of individual quantum dots in high-background environments

Abstract

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