Detailed Information on Publication Record
2001
Theoretical versus experimental resolution in optical microscopy
KOZUBEK, MichalBasic 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
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
20200 2.2 Electrical engineering, Electronic engineering, Information engineering
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 2.165
RIV identification code
RIV/00216224:14330/01:00004204
Organization unit
Faculty of Informatics
UT WoS
000168393000008
Keywords in English
resolution; confocal microscopy; wide-field microscopy; image processing; fluorescent bead
Tags
Tags
International impact, Reviewed
Změněno: 29/1/2021 18:32, prof. RNDr. Michal Kozubek, Ph.D.
Abstract
V originále
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 project |
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| MSM 143300002, plan (intention) |
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| VS97031, research and development project |
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