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@article{362371, author = {Kozubek, Michal}, article_location = {USA}, article_number = {2}, doi = {http://dx.doi.org/10.1002/jemt.1080}, keywords = {resolution; confocal microscopy; wide-field microscopy; image processing; fluorescent bead}, language = {eng}, issn = {1059-910X}, journal = {Microscopy research and technique}, title = {Theoretical versus experimental resolution in optical microscopy}, url = {https://doi.org/10.1002/jemt.1080}, volume = {53}, year = {2001} }
TY - JOUR ID - 362371 AU - Kozubek, Michal PY - 2001 TI - Theoretical versus experimental resolution in optical microscopy JF - Microscopy research and technique VL - 53 IS - 2 SP - 157 EP - 157 PB - Wiley-Liss SN - 1059910X KW - resolution KW - confocal microscopy KW - wide-field microscopy KW - image processing KW - fluorescent bead UR - https://doi.org/10.1002/jemt.1080 L2 - https://doi.org/10.1002/jemt.1080 N2 - The aim of this article is to compare experimental resolution under different conditions with theoretical resolution predicted using electromagnetic diffraction theory. Imaging properties of fluorescent beads of three different diameters (0.1 mum, 0.2 mum, and 0.5 mum) as well as imaging properties of four different fluorescence-stained DNA targets (ABL gene, BCR gene, centromere 6, and centromere 17) are studied. It is shown how the dependence of the resolution on object size varies with wavelength (520 nm versus 580 nm), type of microscopy (wide-field, confocal using Nipkow disk, confocal laser scanning) and basic image processing steps (median and gaussian filters). Furthermore, specimen influence on the resolution was studied (the influence of embedding medium, coverglass thickness, and depth below the coverglass). Both lateral and axial resolutions are presented. The results clearly show that real objects are far from being points and that experimental resolution is often much worse than the theoretical one. Although the article concentrates on fluorescence imaging using high NA objectives, similar dependence can also be expected for other optical arrangements. ER -
KOZUBEK, Michal. Theoretical versus experimental resolution in optical microscopy. \textit{Microscopy research and technique}. USA: Wiley-Liss, 2001, roč.~53, č.~2, s.~157-166. ISSN~1059-910X. Dostupné z: https://dx.doi.org/10.1002/jemt.1080.
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