FandPLimitTool

Single-molecule Localization and Resolution Accuracies

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Introduction

Parameter estimation problems play a central role in single molecule image analysis. Examples include determination of the 2D location of single molecules, estimating the level of defocus of single molecules, determining the distance of separation between two single molecules, estimation of photon count etc. Common to all these problems is that it is helpful for the experimenter to have tools available that can quantify the accuracy with which various parameters can be estimated.

The FandPLimitTool is a GUI based software package that allows users to calculate the limits to the accuracy with which parameters can be estimated from single molecule imaging data. The software supports calculation of limits for the 2D/3D location estimation problem and the 2D/3D distance-estimation/resolution problem. The location estimation problem is concerned with the task of determining the position of a single molecule and the distance-estimation/resolution problem is concerned with the task of determining the distance of separation between two single molecules. The user can calculate limits for a variety of imaging scenarios.

Localization Accuracy

To understand how to use the FandPlimitTool for the calculation of localization accuracy, watch the following YouTube Tutorial (Part I: Localization Accuracy).

Resolution Limit

To understand how to use the FandPlimitTool for the calculation of resolution limits, watch the following YouTube Tutorial (Part II: Resolution Limit).

Download Software

For more information and to dowanload the software, please visit the FandPLimitTool homepage.

  • Software developed in MATLAB (Optimization Toolbox, Image Processing Toolbox, Compiler)
  • Based on object-oriented programming (has a graphical user interface)
  • Add-ons using Autohotkey
  • Hybrid app development (css, html, javascript)

An iPhone app was also developed for this software.

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Related publications

1
SPIE
A. Tahmasbi, S. Ram, J. Chao, A. V. Abraham, E. S. Ward, and R. J. Ober, "An information-theoretic approach to designing the focal plane spacing for multifocal plane microscopy," in Proc. SPIE International Symposium on Biomedical and Optics (BiOS), 9330: 933011, San Francisco, CA, February 11, 2015.
2
QBI
A. Tahmasbi, S. Ram, J. Chao, A. V. Abraham, E. S. Ward, and R. J. Ober, "Mumdesigntool: A software module for designing the plane spacing for multifocal plane microscopy," Quantitative BioImaging 2015, Institut Pasteur, Paris, France, January 7, 2015.
3
IF 3.488
A. Tahmasbi, S. Ram, J. Chao, A. V. Abraham, F. W. Tang, E. S. Ward, and R. J. Ober, "Designing the focal plane spacing for multifocal plane microscopy," Optics Express, vol. 22, no. 14, pp. 16706-16721, 2014.
4
WOL
A. Tahmasbi, "FandPLimitTool and MUMDesignTool user's manual," Ward Ober Lab, University of Texas Southwestern Medical Center, Dallas, USA, 2012.

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