Optical microscope illumination analysis using through-focus scanning optical microscopy

Misalignment of the aperture diaphragm present in optical microscopes results in angular illumination asymmetry (ANILAS) at the sample plane. Here we show that through-focus propagation of ANILAS results in a lateral image shift with a focus position. This could lead to substantial errors in quantit...

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Veröffentlicht in:Optics letters 2017-06, Vol.42 (12), p.2306-2309
Hauptverfasser: Attota, Ravi Kiran, Park, Haesung
Format: Artikel
Sprache:eng
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Zusammenfassung:Misalignment of the aperture diaphragm present in optical microscopes results in angular illumination asymmetry (ANILAS) at the sample plane. Here we show that through-focus propagation of ANILAS results in a lateral image shift with a focus position. This could lead to substantial errors in quantitative results for optical methods that use through-focus images such as three-dimensional nanoparticle tracking, confocal microscopy, and through-focus scanning optical microscopy (TSOM). A correlation exists between ANILAS and the slant in TSOM images. Hence, the slant in the TSOM image can be used to detect, analyze, and rectify the presence of ANILAS.
ISSN:0146-9592
1539-4794
DOI:10.1364/OL.42.002306