Single-Molecule Localization Microscopy  •  Software Benchmarking

Tubulins I

Updated 01 February 2013

This sample consists to a realistic structure of 7 tubulins (constant diameter 25 nm) and 1 tubulin (constant diameter 40 nm).
The depth of the sample is from 0 to 300 nm.
100000 fluorophores are activated over 2400 frames.
Low level of read-out noise autofluorescence background.

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Sequence Download all frames
sequence.zip [181 Mb]

Acquisition Parameters

Camera
Photon converter factor or Quantum efficiency (QE)1.00e-/Ph.
QE = QE-Gain x QE-interacting [Ref. 1]
Resolution256pixels
Pixelsize150.00nm
Field of view38400.00nm
Optics
Wavelength723.00nm
Numerical aperture (NA)1.40
Diffraction limit258.21nm
Drift
DeriveDisable
ShakingDisable
Analog Digital Conversion
Electron conversion - Gain1.00DN/e-
Electron conversion - Offset0.00DN
Baseline100.00DN
Saturation16383.00DN
Quantization14-bit

Reference

[1] James R. Janesick, Photon Transfer, SPIE Press Monograph Vol. PM170, 2007.

Point-Spread Function (PSF)

Physical Model
Wavelength723.00nm
Numerical aperture (NA)1.40
Diffraction limit258.21nm
Offset focal plane 0nm
XY functionGaussian
FWHM258.21nm
Z functionExponential
Focal plane1 x FWHM at 0.0 nm
Defocus plane2 x FWHM at 500.0 nm

Reference

[1] H. Kirshner, F. Aguet, D. Sage, M. Unser, "3-D PSF Fitting for Fluorescence Microscopy: Implementation and Localization Applications", Journal of Microscopy, in press.

[2] PSF Generator An ImageJ plugin to generate 3D microscope Point-Spread Function (PSF).

© 2017 Biomedical Imaging Group, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
Last update: 31 Mar 2017