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Appl Opt. 2006 Jul 10;45(20):5037-45.

High-resolution image reconstruction in fluorescence microscopy with patterned excitation.

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  • 1Randall Division of Cell and Molecular Biophysics, Kings College London, London SE1 1UL, UK. rainer.heintzmann@kcl.ac.uk

Abstract

We discuss data treatment strategies in structured illumination microscopy, using simulated and experimental data. In the setup, the illumination pattern is generated by projecting a movable pinhole mask into the sample, and a wide-field fluorescence microscope image is acquired for each pattern position. The structured illumination data obtained from a two-dimensional illumination pattern can be treated by projection strategies such as in video confocal microscopy (sum, maximum, maximum minus minimum, and superconfocal), by a scaled subtraction of the out-of-focus estimate, or by a modified version of the Fourier-space treatment, as is known for data from one-dimensional structured illumination. We investigate the influence of some data processing strategies on unwanted effects such as residual patterning and local deviations from linearity in the reconstructed intensity.

PMID:
16807615
[PubMed - indexed for MEDLINE]
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