Open AccessThree dimensional time-gated tracking of non-blinking quantum dots in live cells
Author(s) -
Matthew S. DeVore,
Dominik Stich,
Aaron M. Keller,
Yagnaseni Ghosh,
Peter M. Goodwin,
Mary E. Phipps,
Michael H. Stewart,
Cédric Cleyrat,
Bridget S. Wilson,
Diane S. Lidke,
Jennifer A. Hollingsworth,
James H. Werner
Publication year - 2015
Publication title -
proceedings of spie, the international society for optical engineering/proceedings of spie
Language(s) - English
Resource type - Conference proceedings
SCImago Journal Rank - 0.192
H-Index - 176
eISSN - 1996-756X
pISSN - 0277-786X
DOI - 10.1117/12.2082943
Subject(s) - tracking (education) , quantum dot , microscope , signal (programming language) , noise (video) , microscopy , computer science , optoelectronics , biological system , materials science , computer vision , optics , physics , psychology , pedagogy , image (mathematics) , biology , programming language
Single particle tracking has provided a wealth of information about biophysical processes such as motor protein transport and diffusion in cell membranes. However, motion out of the plane of the microscope or blinking of the fluorescent probe used as a label generally limits observation times to several seconds. Here, we overcome these limitations by using novel non-blinking quantum dots as probes and employing a custom 3D tracking microscope to actively follow motion in three dimensions (3D) in live cells. Signal-to-noise is improved in the cellular milieu through the use of pulsed excitation and time-gated detection.