Three-dimensional particle tracking via bifocal imaging.
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.
Toprak
Erdal
E
Department of Biophysics, University of Illinois Urbana Champaign, Urbana, Illinois 61801, USA.
Balci
Hamza
H
Blehm
Benjamin H
BH
Selvin
Paul R
PR
D002460:Cell Line; D006801:Humans; D021621:Imaging, Three-Dimensional; D008863:Microspheres