1999
High-resolution cytometry of FISH dots in interphase cell nuclei
KOZUBEK, Michal, Stanislav KOZUBEK, Emilie LUKÁŠOVÁ, Andrea MAREČKOVÁ, Eva BÁRTOVÁ et. al.Základní údaje
Originální název
High-resolution cytometry of FISH dots in interphase cell nuclei
Autoři
KOZUBEK, Michal (203 Česká republika), Stanislav KOZUBEK (203 Česká republika), Emilie LUKÁŠOVÁ (203 Česká republika), Andrea MAREČKOVÁ, Eva BÁRTOVÁ (203 Česká republika), Magdalena SKALNÍKOVÁ (203 Česká republika) a Adriana JERGOVÁ (203 Česká republika)
Vydání
Cytometry, New York, International Society for Analytical Cyt, 1999, 0196-4763
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
20200 2.2 Electrical engineering, Electronic engineering, Information engineering
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 2.843
Kód RIV
RIV/00216224:14330/99:00001446
Organizační jednotka
Fakulta informatiky
UT WoS
000081578600002
Klíčová slova anglicky
fluorescence in situ hybridization; interphase nuclei; fluorescence microscopy; automated microscopy; image analysis; 3-D analysis; high-resolution cytometry
Štítky
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 7. 5. 2010 17:12, prof. RNDr. Michal Kozubek, Ph.D.
Anotace
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.
Návaznosti
| GA202/97/0874, projekt VaV |
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| GA202/99/P008, projekt VaV |
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| MSM 143300002, záměr |
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| VS97031, projekt VaV |
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