title: Three-dimensional particle tracking via bifocal imaging.
abstract: We introduce a bifocal imaging method that enables three-dimensional (3D) tracking of both fluorescent and nonfluorescent particles. We accomplish this by simultaneously imaging a focused plane, for in-plane position (x,y), and a defocused plane, for out-of-plane position (z), of a molecule using a single CCD camera. We applied our method to several systems including in vivo melanosome tracking and phagocytosed fluorescent bead tracking. We have achieved 2-5 nm accuracy with a 2-50 ms time resolution.
authors.affiliation: Department of Biophysics, University of Illinois Urbana Champaign, Urbana, Illinois 61801, USA.
authors.forename: Erdal
authors.initials: E
authors.lastname: Toprak
authors.forename: Hamza
authors.initials: H
authors.lastname: Balci
authors.forename: Benjamin H
authors.initials: BH
authors.lastname: Blehm
authors.forename: Paul R
authors.initials: PR
authors.lastname: Selvin
mesh_terms: D002460:Cell Line; D006801:Humans; D021621:Imaging, Three-Dimensional; D008863:Microspheres