GUI for fitting 3D single-molecule images using the double-helix PSF
This package of MATLAB routines with accompanying GUI allows scientific users to measure the 3D location of single molecules when using the double-helix point spread function (DH-PSF) widefield microscope. Version 2 also includes code that facilitates combining localization data from two spectral channels using a locally-weighted quadratic 3D registration function
Features
- Double-Gaussian estimator is calibrated via an axial scan of bright immobile fluorescent emitters (e.g. beads)
- DH localization is performed through template matching and subsequent double Gaussian fitting
- Tiff stacks of SM images are analyzed using template matching followed by double-Gaussian fitting to extract estimates of the molecule positions
- Two spectral channels are registered by generating a locally-weighted quadratic mapping function from control point pairs
SPIW is a MATLAB toolbox, for automated processing of scanning probe microscope images. Tools are applicable to all SPM images, but the main focus is on images with atomic or molecular resolution.
SPIW is primarily a MATLAB toolbox with functions designed to be called in a user's own script. SPIW also has a very basic GUI for browsing and exporting images.
For information on SPIW's performance see our publication ( http://dx.doi.org/10.1063/1.4827076 ) in the Review of Scientific Instruments.
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This toolkit allows complete control of a microscopy setup from Labview, Matlab, Scilab, Python, .Net, VB, IgorPro, Mathematica and more. Included is a standalone program for image acquisition and scripting control of a scientific microscope.