Open AccessDual-color multiple-particle tracking at 50-nm localization and over 100-µm range in 3D with temporal focusing two-photon microscopy
Author(s) -
Yu Ding,
Chunqiang Li
Publication year - 2016
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.7.004187
Subject(s) - tracking (education) , particle (ecology) , microscopy , photon , optics , fluorescence , range (aeronautics) , particle tracking velocimetry , physics , materials science , two photon excitation microscopy , psychology , particle image velocimetry , pedagogy , turbulence , oceanography , composite material , thermodynamics , geology
Nanoscale particle tracking in three dimensions is crucial to directly observe dynamics of molecules and nanoparticles in living cells. Here we present a three-dimensional particle tracking method based on temporally focused two-photon excitation. Multiple particles are imaged at 30 frames/s in volume up to 180 × 180 × 100 µm 3 . The spatial localization precision can reach 50 nm. We demonstrate its capability of tracking fast swimming microbes at speed of ~200 µm/s. Two-photon dual-color tracking is achieved by simultaneously exciting two kinds of fluorescent beads at 800 nm to demonstrate its potential in molecular interaction studies. Our method provides a simple wide-field fluorescence imaging approach for deep multiple-particle tracking.