Detailed Information on Publication Record
1999
High-resolution cytometry of FISH dots in interphase cell nuclei
KOZUBEK, Michal, Stanislav KOZUBEK, Emilie LUKÁŠOVÁ, Andrea MAREČKOVÁ, Eva BÁRTOVÁ et. al.Basic information
Original name
High-resolution cytometry of FISH dots in interphase cell nuclei
Authors
KOZUBEK, Michal (203 Czech Republic), Stanislav KOZUBEK (203 Czech Republic), Emilie LUKÁŠOVÁ (203 Czech Republic), Andrea MAREČKOVÁ, Eva BÁRTOVÁ (203 Czech Republic), Magdalena SKALNÍKOVÁ (203 Czech Republic) and Adriana JERGOVÁ (203 Czech Republic)
Edition
Cytometry, New York, International Society for Analytical Cyt, 1999, 0196-4763
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
Impact factor
Impact factor: 2.843
RIV identification code
RIV/00216224:14330/99:00001446
Organization unit
Faculty of Informatics
UT WoS
000081578600002
Keywords in English
fluorescence in situ hybridization; interphase nuclei; fluorescence microscopy; automated microscopy; image analysis; 3-D analysis; high-resolution cytometry
Tags
Tags
International impact, Reviewed
Změněno: 7/5/2010 17:12, prof. RNDr. Michal Kozubek, Ph.D.
Abstract
V originále
Background. Flow cytometry (FCM) and laser scanning cytometry (LSCM) provide indispensable tools for measuring large number of cells with low resolution. Confocal microscopy, on the other hand, is used for measuring small number of cells with high resolution. In this paper, we present a reasonable compromise between the two extremes. Methods. We have developed a completely automated, high-resolution system (high-resolution cytometer, HRCM) capable of analyzing microscope slides with FISH-stained interphase nuclei in 2-D as well as in 3-D using a fully motorized epi-fluorescence microscope and a cooled digital CCD camera fully controlled by a high-performance computer which performs both acquisition and related on-line image analysis. The images of different dyes are acquired sequentially using highly-specific filters and superimposed in computer memory. For each nucleus and each hybridization dot, user-selected attributes (such as position, size, intensity, etc.) are computed off-line using another processor or computer connected with a network. Results. Using HRCM, it is possible to analyze multi-color preparations including UV-excited dyes as well as repeatedly hybridized preparations re-acquiring individual nuclei. The speed of the acquisition and analysis is about 50 nuclei per minute in 2-D and 1 nucleus per minute in 3-D but depends on the density of nuclei on the slide; the precision of the lateral and axial measurements is approximately 100 nm. Conclusions. Thus, using overnight acquisition, quantities comparable to those of FCM or LSCM measurements can be analyzed with an accuracy comparable to confocal microscopy. HRCM is suitable for a number of clinical and scientific tasks: routine diagnostics, follow-up of therapy, studies of chromatin structure and many other different aspects of cell research.
Links
| GA202/97/0874, research and development project |
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| 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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