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3‐Dimensional Tracking of Non‐blinking ‘Giant’ Quantum Dots in Live Cells
Author(s) -
Keller Aaron M.,
Ghosh Yagnaseni,
DeVore Matthew S.,
Phipps Mary E.,
Stewart Michael H.,
Wilson Bridget S.,
Lidke Diane S.,
Hollingsworth Jennifer A.,
Werner James H.
Publication year - 2014
Publication title -
advanced functional materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.069
H-Index - 322
eISSN - 1616-3028
pISSN - 1616-301X
DOI - 10.1002/adfm.201400349
Subject(s) - quantum dot , tracking (education) , materials science , intermittency , microscopy , particle (ecology) , photonics , nanotechnology , biological system , optoelectronics , physics , optics , biology , thermodynamics , psychology , ecology , pedagogy , turbulence
While semiconductor quantum dots (QDs) have been used successfully in numerous single particle tracking (SPT) studies due to their high photoluminescence efficiency, photostability, and broad palette of emission colors, conventional QDs exhibit fluorescence intermittency or ‘blinking,’ which causes ambiguity in particle trajectory analysis and limits tracking duration. Here, non‐blinking ‘giant’ quantum dots (gQDs) are exploited to study IgE‐FcεRI receptor dynamics in live cells using a confocal‐based 3D SPT microscope. There is a 7‐fold increase in the probability of observing IgE‐FcεRI for longer than 1 min using the gQDs compared to commercially available QDs. A time‐gated photon‐pair correlation analysis is implemented to verify that selected SPT trajectories are definitively from individual gQDs and not aggregates. The increase in tracking duration for the gQDs allows the observation of multiple changes in diffusion rates of individual IgE‐FcεRI receptors occurring on long (>1 min) time scales, which are quantified using a time‐dependent diffusion coefficient and hidden Markov modeling. Non‐blinking gQDs should become an important tool in future live cell 2D and 3D SPT studies, especially in cases where changes in cellular dynamics are occurring on the time scale of several minutes.