Multidimensional Systems and Signal Processing

, Volume 17, Issue 1, pp 27–57

A Stochastic Analysis of Performance Limits for Optical Microscopes

Authors

  • Sripad Ram
    • Center for Immunology, NB9.106University of Texas Southwestern Medical Center at Dallas
    • Joint Biomedical Engineering Graduate ProgramUniversity of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas
  • E. Sally Ward
    • Center for Immunology, NB9.106University of Texas Southwestern Medical Center at Dallas
    • Center for Immunology, NB9.106University of Texas Southwestern Medical Center at Dallas
    • Department of Electrical EngineeringUniversity of Texas at Dallas Richardson
Original Article

DOI: 10.1007/s11045-005-6237-2

Cite this article as:
Ram, S., Sally Ward, E. & Ober, R.J. Multidim Syst Sign Process (2006) 17: 27. doi:10.1007/s11045-005-6237-2

Abstract

The optical microscope is a powerful instrument for observing cellular events. Recently, the increased use of microscopy in quantitative biological research, including single molecule microscopy, has generated significant interest in determining the performance limits of an optical microscope. Here, we formulate this problem in the context of a parameter estimation approach in which the acquired imaging data is modeled as a spatio-temporal stochastic process. We derive formulations of the Fisher information matrix for models that allow both stationary and moving objects. The effects of background signal, detector size, pixelation and noise sources are also considered. Further, formulations are given that allow the study of defocused objects. Applications are discussed for the special case of the estimation of the location of objects, especially single molecules. Specific emphasis is placed on the derivation of conditions that guarantee block diagonal or diagonal Fisher information matrices.

Keywords

Spatio-temporal stochastic processesFisher information matrixCramer-Rao lower boundParameter estimationFluorescence microscopyOptical imagingSingle Molecule MicroscopyLocalization accuracy

Copyright information

© Springer Science + Business Media, Inc. 2006