UNIVERSITY OF BUCHAREST
FACULTY OF PHYSICS

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2024-11-22 2:18
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Conference: Bucharest University Faculty of Physics 2018 Meeting


Section: Biophysics; Medical Physics


Title:
Localizer for Igor as a drift correction tool in STORM Microscopy


Authors:
Andreea Ioana CIOCĂNARU (1), Claudia CHILOM (1), Karmen ABUZINEH (2), Maram ABADI (2), Satoshi HABUCHI (2)


Affiliation:
1) Faculty of Physics, University of Bucharest

2) King Abdullah University of Science and Technology


E-mail
andreea.c@protonmail.com


Keywords:
STORM, localization, drift correction, fluorescnce microscopy


Abstract:
For a long time it has been thought that optical microscopy will never obtain a better resolution than half the light wavelength, which is typically about 200 nm. It is still the case today that light microscopy is the most popular technique to be used in life sciences because it has the ability to non-destructively images specific molecules in real time. The process of sample imaging involves steps that might lead to propagating errors. One of the issues that can be encountered during imaging is sample instability due to factors such as the use of immersion oil, changes in temperature or mechanical drift. Stochastic Optical Reconstruction Microscopy (STORM) uses subsets of sparse fluorescent molecules that switch from an off to an on-state to reconstruct a high resolution image. We have been working on qualitatively comparing the outcome of two drift correction protocols using Localizer for Igor software to analyze STORM data. ImageJ was used to evaluate the colocalizations of the two protocols in order to evaluate their effectiveness. Results show that the Subimages protocol returned more localizations than the ad-hoc fiducials protocol, using the same threshold and fitting conditions. Furthermore, we aim to estimate the relative position of fluorescent molecules on the KG1a cell membrane using gold nanoparticles as a surface marker.


References:

1. Heilemann M. et al. Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes. Angew Chem-Int. Edit, 47, 6172-6176, 2008

2. Huang B., Wang W. Q., Bates M. & Zhuang X. W. Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy. Science, 319, 810-813, 2008