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001-es BibID:	BIBFORM050042
035-os BibID:	PMID:23504771
Első szerző:	Fábián Ákos István (aneszteziológus)
Cím:	TripleFRET measurements in flow cytometry / Ákos Fábián, Gábor Horváth, György Vámosi, György Vereb, János Szöllősi
Dátum:	2013
Megjegyzések:	A frequently used method for viewing protein interactions and conformation, Forster (fluorescence) resonance energy transfer (FRET), has traditionally been restricted to two fluorophores. Lately, several methods have been introduced to expand FRET methods to three species. We present a method that allows the determination of FRET efficiency in three-dye systems on a flow cytometer. TripleFRET accurately reproduces energy transfer efficiency values measured in two-dye systems, and it can indicate the presence of trimeric complexes, which is not possible with conventional FRET methods. We also discuss the interpretation of energy transfer values obtained with tripleFRET in relation to spatial distribution of labeled molecules, specifically addressing the limitations of using total energy transfer to determine molecular distance
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban analysis Antibodies Antibodies,Monoclonal,Humanized article Biophysics cell biology Cell Line chemistry cytometry Dyes Energy Transfer ENERGY-TRANSFER FLOW Flow Cytometry Fluorescence Fluorescence Resonance Energy Transfer fluorescent dye Fluorescent Dyes FRET Humans Hungary Immunoconjugates immunology methods Protein Binding PROTEIN INTERACTIONS Research Research Support resonance energy transfer Staining and Labeling statistics & numerical data Support
Megjelenés:	Cytometry Part A. - 83 : 4 (2013), p. 375-385. -
További szerzők:	Horváth Gábor (1974-) (biofizikus) Vámosi György (1967-) (biofizikus) Vereb György (1965-) (biofizikus, orvos) Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:	NK 101337 OTKA K75752 OTKA TÁMOP-4.2.2-08/1-2008-0019 TÁMOP TÁMOP-4.2.1/B-09/1/KONV-2010-0007 TÁMOP Receptor tirozin kinázok mint terápiás célpontok : működésük szabályozásának, és a közöttük fellépő molekuláris kölcsönhatások vizsgálata TÁMOP-4.2.2.A-11/1/KONV-2012-0025 TÁMOP Baross Gábor Program REG_EA_09-1-2009-0010 Egyéb K77600 OTKA K103965 OTKA
Internet cím:	DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM042654
035-os BibID:	(scopus)19444380683 (wos)000229353200007
Első szerző:	Horváth Gábor (biofizikus)
Cím:	Selecting the right fluorophores and flow cytometer for fluorescence resonance energy transfer measurements / Gábor Horváth, Miklós Petrás, Gergely Szentesi, Ákos Fábián, John W. Park, György Vereb, János Szöllősi
Dátum:	2005
ISSN:	1552-4922 1552-4930
Megjegyzések:	BACKGROUND:Fluorescence resonance energy transfer applied in flow cytometry (FCET) is an excellent tool for determining supramolecular organization of biomolecules at the cell surface or inside the cell. Availability of new fluorophores and cytometers requires the establishment of fluorophore dye pairs most suitable for FCET measurements.METHODS:A gastric tumor cell line (N87) was labeled for major histocompatibility complex class I heavy chain and beta2-microglobulin with antibodies conjugated with fluorescein- and indocarbocyanine-like fluorophores and analyzed in FCET measurements on a cell-by-cell basis using three flow cytometers: FACSCalibur, FACSDiVa, and FACSArray.RESULTS:Normalized fluorescence intensity values were measured and normalized energy transfer efficiencies, spectral overlap integrals, and crucial dye- and instrument-dependent parameters were calculated for all matching pairs of seven fluorophores on the three commercial cytometers. The most crucial parameter in determining the applicability of the donor-acceptor pairs was the normalized fluorescence intensity and the least important one was the spectral overlap.CONCLUSIONS:On the basis of available laser lines, the optimal dye pair for all three cytometers is the Alexa546-Alexa647 pair, which produces high energy transfer efficiency values and has the best spectral characteristics with regard to laser excitation, detection of emission, and spectral overlap.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk fluorescence resonance energy transfer flow cytometry donor-acceptor pair egyetemen (magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A. - 65A : 2 (2005), p. 148-157. -
További szerzők:	Petrás Miklós (1977-) (orvos) Szentesi Gergely (1976-) (kémia-fizika tanár) Fábián Ákos István (1982-) (aneszteziológus) Park, John W. Vereb György (1965-) (biofizikus, orvos) Szöllősi János (1953-) (biofizikus)
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM004566
Első szerző:	Kovács Tamás (szülész-nőgyógyász, humángenetikus)
Cím:	DNA flow cytometry of human spermatozoa : consistent stoichiometric staining of sperm DNA using a novel decondensation protocol / Kovács T., Békési G., Fábián A., Rákosy Z., Horváth G., Mátyus L., Balázs M., Jenei A.
Dátum:	2008
Megjegyzések:	Rapid flow cytometric measurement of the frequency of aneuploid human sperms is in increasing demand but development of an exploitable method is hindered by difficulties of stoichiometric staining of sperm DNA. An aggressive decondensation protocol is needed after which cell integrity still remains intact. We used flow cytometry to examine the effect of lithium diiodosalicylate (LIS, chaotropic agent) on fluorescence intensity of propidium iodide-treated human spermatozoa from 10 normozoospermic men. When flow cytometric identification of diploid spermatozoa was achieved, validation was performed after sorting by three-color FISH. In contrast with the extremely variable histograms of nondecondensed sperms, consistent identification of haploid and diploid spermatozoa was possible if samples were decondensed with LIS prior to flow cytometry. A 76-fold enrichment of diploid sperms was observed in the sorted fractions by FISH. A significant correlation was found between the proportion of sorted cells and of diploid sperms by FISH. Application of LIS during the preparation of sperm for flow cytometry appears to ensure the stoichiometric staining of sperm DNA, making quantification of aneuploid sperm percentage possible. To our knowledge this is the first report in terms of separating spermatozoa with confirmedly abnormal chromosomal content. High correlation between the proportion of cells identified as having double DNA content by flow cytometry and diploid sperm by FISH allows rapid calculation of diploidy rate. Copyright 2008 International Society for Advancement of Cytometry.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban diploid spermatozoa aneuploid spermatozoa propidium iodide lithium diiodosalicylate egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A 73 : 10 (2008), p. 965-970. -
További szerzők:	Békési Gyöngyi Fábián Ákos István (1982-) (aneszteziológus) Rákosy Zsuzsa (1978-) (sejtbiológus, molekuláris biológus, genetikus) Horváth Gábor (1974-) (biofizikus) Mátyus László (1956-) (biofizikus) Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Jenei Attila (1966-) (biofizikus)
Internet cím:	elektronikus változat DOI elektronikus változat
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001-es BibID:	BIBFORM004886
Első szerző:	Szentesi Gergely (kémia-fizika tanár)
Cím:	Computer program for analyzing donor photobleaching FRET image series / Szentesi, G., Vereb, G., Horvath, G., Bodnar, A., Fabian, A., Matko, J., Gaspar, R., Damjanovich, S., Matyus, L., Jenei, A.
Dátum:	2005
ISSN:	1552-4922
Megjegyzések:	The photobleaching fluorescence resonance energy transfer (pbFRET) technique is a spectroscopic method to measure proximity relations between fluorescently labeled macromolecules using digital imaging microscopy. To calculate the energy transfer values one has to determine the bleaching time constants in pixel-by-pixel fashion from the image series recorded on the donor-only and donor and acceptor double-labeled samples. Because of the large number of pixels and the time-consuming calculations, this procedure should be assisted by powerful image data processing software. There is no commercially available software that is able to fulfill these requirements. METHODS: New evaluation software was developed to analyze pbFRET data for Windows platform in National Instrument LabVIEW 6.1. This development environment contains a mathematical virtual instrument package, in which the Levenberg-Marquardt routine is also included. As a reference experiment, FRET efficiency between the two chains (beta2-microglobulin and heavy chain) of major histocompatibility complex (MHC) class I glycoproteins and FRET between MHC I and MHC II molecules were determined in the plasma membrane of JY, human B lymphoma cells. RESULTS: The bleaching time constants calculated on pixel-by-pixel basis can be displayed as a color-coded map or as a histogram from raw image format. CONCLUSION: In this report we introduce a new version of pbFRET analysis and data processing software that is able to generate a full analysis pattern of donor photobleaching image series under various conditions.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Algorithms analysis beta 2-Microglobulin Biophysics Cell Line,Tumor Cells Energy Transfer Fluorescence Fluorescence Resonance Energy Transfer Glycoproteins Histocompatibility Antigens Human Humans Hungary Lymphoma Major Histocompatibility Complex metabolism methods Microscopy Photobleaching Research Software Support egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A. - 67 : 2 (2005), p. 119-128. -
További szerzők:	Vereb György (1965-) (biofizikus, orvos) Horváth Gábor (1974-) (biofizikus) Dóczy-Bodnár Andrea (1970-) (biofizikus) Fábián Ákos István (1982-) (aneszteziológus) Matkó János (1952-) (biológus) Gáspár Rezső (1944-) (biofizikus) Damjanovich Sándor (1936-2017) (biofizikus) Mátyus László (1956-) (biofizikus) Jenei Attila (1966-) (biofizikus)
Internet cím:	elektronikus változat DOI
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