KOZUBEK, Michal. Theoretical versus experimental resolution in optical microscopy. Microscopy research and technique. USA: Wiley-Liss, 2001, vol. 53, No 2, p. 157-166. ISSN 1059-910X. Available from: https://dx.doi.org/10.1002/jemt.1080.
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Basic information
Original name Theoretical versus experimental resolution in optical microscopy
Authors KOZUBEK, Michal (203 Czech Republic, guarantor, belonging to the institution).
Edition Microscopy research and technique, USA, Wiley-Liss, 2001, 1059-910X.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 20200 2.2 Electrical engineering, Electronic engineering, Information engineering
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 2.165
RIV identification code RIV/00216224:14330/01:00004204
Organization unit Faculty of Informatics
Doi http://dx.doi.org/10.1002/jemt.1080
UT WoS 000168393000008
Keywords in English resolution; confocal microscopy; wide-field microscopy; image processing; fluorescent bead
Tags cbia-web, confocal microscopy, fluorescent bead, image processing, RESOLUTION, wide-field microscopy
Tags International impact, Reviewed
Changed by Changed by: prof. RNDr. Michal Kozubek, Ph.D., učo 3740. Changed: 29/1/2021 18:32.
Abstract
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.
Links
GA202/99/P008, research and development projectName: Využití analýzy obrazu při studiu struktury interfázního jádra
Investor: Czech Science Foundation, Image analysis in the study of interphase nucleus structure
MSM 143300002, plan (intention)Name: Využití počítačové analýzy obrazu v optické mikroskopii
Investor: Ministry of Education, Youth and Sports of the CR, Application of computer image analysis in optical microscopy
VS97031, research and development projectName: Využití analýzy obrazu při studiu mechanismů vzniku, v diagnostice a pro prevenci závažných onemocnění člověka
Investor: Ministry of Education, Youth and Sports of the CR, Image analysis in the study of mechanisms of induction, in diagnosis and for prevention of deleterious human diseases
PrintDisplayed: 21/6/2024 18:35