title: Dual-color multiple-particle tracking at 50-nm localization and over 100-µm range in 3D with temporal focusing two-photon microscopy.
abstract: 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 µm3. 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.
authors.affiliation: Department of Physics, The University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Department of Physics, The University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, USA; Border Biomedical Research Center, The University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, USA.
authors.forename: Chunqiang
authors.initials: C
authors.lastname: Li
keywords: (170.2520) Fluorescence microscopy; (170.7160) Ultrafast technology; (180.4315) Nonlinear microscopy