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Proceedings Article

Point spread function optimization for STORM using adaptive optics

[+] Author Affiliations
Kayvan Forouhesh Tehrani, Peter Kner

The Univ. of Georgia (United States)

Proc. SPIE 8978, MEMS Adaptive Optics VIII, 89780I (March 7, 2014); doi:10.1117/12.2040390
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From Conference Volume 8978

  • MEMS Adaptive Optics VIII
  • Thomas G. Bifano; Joel Kubby; Sylvain Gigan
  • San Francisco, California, United States | February 01, 2014

abstract

Stochastic Optical Reconstruction Microscopy (STORM) requires a high Strehl ratio point spread function (PSF) to achieve high resolution, especially in the presence of background fluorescence. The PSF is degraded by aberrations caused by imperfections in the optics, the refractive index mismatch between the sample and coverslip, and the refractive index variations of the sample. These aberrations distort the shape of the PSF and increase the PSF width directly reducing the resolution of STORM. Here we discuss the use of Adaptive Optics (AO) to correct aberrations, maintaining a high Strehl ratio even in thick tissue. Because the intensity fluctuates strongly from frame to frame, image intensity is not a reliable measure of PSF quality, and the choice of a robust optimization metric is critical. We demonstrate the use of genetic algorithms with single molecule imaging for optimization of the wavefront and introduce a metric that is relatively insensitive to image intensity. We demonstrate the correction of the wavefront from measurements of single quantum dots. © (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Kayvan Forouhesh Tehrani and Peter Kner
" Point spread function optimization for STORM using adaptive optics ", Proc. SPIE 8978, MEMS Adaptive Optics VIII, 89780I (March 7, 2014); doi:10.1117/12.2040390; http://dx.doi.org/10.1117/12.2040390


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