3D single molecule tracking and superresolution microscopy using multifocal plane microscopy.
The study of cellular processes in three-dimensions is severely limited by the lack of imaging methodologies that allow for fast 3D tracking of cellular events and 3D superresolution imaging of sub-cellular structures. We have developed a 3D imaging modality, multifocal plane microscopy (MUM), that provides a powerful approach for 3D single molecule tracking and 3D superresolution microscopy. Here we review the technical challenges associated with 3D single molecule localization that is fundamental to both 3D tracking and 3D superresolution, and discuss how MUM overcomes these problems.
Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Sripad
S
Ram
Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
E Sally
ES
Ward
Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX, USA; Department of Electrical Engineering, University of Texas at Dallas, Richardson, TX, USA.
Raimund J
RJ
Ober
2D/3D localization accuracy; Cramer-Rao lower bound; Fisher information matrix; maximum likelihood estimator; resolution limits