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1.

001-es BibID:BIBFORM046090
Első szerző:Bacsó Zsolt (biofizikus)
Cím:Raft and cytoskeleton associations of an ABC transporter : P-glycoprotein / Zsolt Bacso, Henrietta Nagy, Katalin Goda, László Bene, Ferenc Fenyvesi, János Matkó, Gábor Szabó
Dátum:2004
ISSN:0196-4763
Megjegyzések:A novel flow cytometric assay has been described in an accompanying report (Gombos et al., METHODS: The kinetics of the decrease in immunofluorescence intensity was analyzed after the addition of the raft-preserving Triton X-100 or Nonidet P-40, both of which disrupt the entire membrane. Mild treatments by both detergents leave cells attached to only those proteins that are anchored to the cytoskeleton by rafts or independent of rafts. Agents that affect microfilaments and modulate membrane levels of cholesterol by cyclodextrin were used to distinguish between the raft-mediated and non-raft-related associations of the Pgp. Confocal microscopy and flow cytometric fluorescence energy transfer measurements were used to confirm colocalization of Pgp with raft constituents. RESULTS: The assay was proved to be sensitive enough to resolve differences between the resistance of UIC2-labeled cell-surface Pgps to Triton X-100 versus Nonidet P-40. Approximately 34% of the UIC2 Fab-labeled Pgp molecules were associated with the cytoskeleton through detergent-resistant, cholesterol-sensitive microdomains or directly, whereas approximately 15% were found to be directly linked to the cytoskeleton. Accordingly, confocal microscopy showed that Pgps colocalize with raft markers, mainly in microvilli. Fluorescence resonance energy transfer efficiency data indicating molecular proximity between Pgp and the raft markers CD44, CD59, and G(M1)-gangliosides also suggested that a significant fraction of Pgps resides in raft microdomains. Raft association of Pgp appears to be of functional significance because its modulation markedly affected drug pumping. CONCLUSIONS: By using the flow cytometric detergent resistance assay in kinetic mode, we were able to assess the extent of raft association and actin cytoskeleton anchorage of Pgp expressed at physiologically relevant levels. We demonstrated that a significant fraction of Pgp is raft associated on LS-174-T human colon carcinoma cells and that this localization may influence its transporter function. The kinetic flow cytometric detergent resistance assay presented in this report is considered to be generally applicable for the analysis of molecular interactions of membrane proteins expressed at low levelsújratöltve - BIBFORM004828
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:Cytometry. - 61 : 2 (2004), p. 105-116. -
További szerzők:Nagy Henrietta Goda Katalin (1969-) (biofizikus) Bene László (1963-) (biofizikus) Fenyvesi Ferenc (1977-) (gyógyszerész, gyógyszertechnológus) Matkó János (1952-) (biológus) Szabó Gábor (1953-) (biofizikus)
Pályázati támogatás:T032563
OTKA
034393
OTKA
T046945
OTKA
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DOI
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2.

001-es BibID:BIBFORM106370
035-os BibID:(scopus)85145182203 (wos)000903915900001
Első szerző:Bene László (biofizikus)
Cím:Deep-learning FRET visualization in flow cytometry : at the cross road of the signaling and FRET pathways / Bene László, Damjanovich László
Dátum:2022
ISSN:1552-4922 1552-4930
Megjegyzések:Present day flow cytometers and microscopes enable combined signal detection as a function of many parameters such as the color and polarization of the exciting and emitted light, and the spatial position and orientation of the detected object, as well as the time point of the data registration.
Tárgyszavak:Orvostudományok Elméleti orvostudományok hozzászólás
folyóiratcikk
artificial neural network
convolutional neural network
reduction of dimensionality
signal multiplexing
supervised and unsupervised deep-learning
Megjelenés:Cytometry Part A. - 12 (2022), p. 1-7. -
További szerzők:Damjanovich László (1960-) (általános sebész)
Pályázati támogatás:TÁMOP-4.2.2.A-11/1/KONV-2012-0045
TÁMOP
OSTRAT/ 810/213
OTKA
1G3DBLR0TUDF-247
Egyéb
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3.

001-es BibID:BIBFORM102133
035-os BibID:(WOS)000788442500001 (Scopus)85128962098
Első szerző:Bene László (biofizikus)
Cím:Spectral flow cytometric FRET : Towards a hyper dimensional flow cytometry / Bene László, Damjanovich László
Dátum:2022
ISSN:1552-4922
Tárgyszavak:Orvostudományok Klinikai orvostudományok hozzászólás
folyóiratcikk
biochemical resolution
Fisher information
fluorescence lifetime
fluorescence polarization
anisotropy
kinase reporter assay
signal multiplexing
spectral phasor analysis
Megjelenés:Cytometry Part A. - 101 : 6 (2022), p. 468-473. -
További szerzők:Damjanovich László (1960-) (általános sebész)
Pályázati támogatás:TÁMOP-4.2.2.A-11/1/KONV-2012-0045
TÁMOP
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4.

001-es BibID:BIBFORM090876
035-os BibID:(WoS)000569478200001 (Scopus)85090924650
Első szerző:Bene László (biofizikus)
Cím:When the Complex Makes It Easy : phasor Plotting as a Model Independent Representation of Fluorescence Decay in Flow Cytometry / Bene László, Damjanovich László
Dátum:2020
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry Part A. - 97 : 12 (2020), p. 1211-1216. -
További szerzők:Damjanovich László (1960-) (általános sebész)
Pályázati támogatás:TAMOP-4.2.2.A-11/1/KONV-2012-0045
TÁMOP
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5.

001-es BibID:BIBFORM082467
035-os BibID:(Wos)000482423800001 (Scopus)85065183846
Első szerző:Bene László (biofizikus)
Cím:Förster Resonance Energy Transfer Pioneers Biomechanics : molecular-Scale Force Meters for Visualizing Intracellular Stress Fields / László Bene, László Damjanovich
Dátum:2019
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry Part A. - 95 : 8 (2019), p. 819-822. -
További szerzők:Damjanovich László (1960-) (általános sebész)
Pályázati támogatás:TÁMOP-4.2.2.A-11/1/KONV-2012-0045
TÁMOP
Sebészet Kutatócsoport
Internet cím:DOI
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6.

001-es BibID:BIBFORM057536
Első szerző:Bene László (biofizikus)
Cím:À la Fizeau in flow : pulse shape-assisted fluorescence lifetime / László Bene, János Szöllősi
Dátum:2014
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok hozzászólás
folyóiratcikk
fluorescence
resonance
energy
transfer
Megjelenés:Cytometry Part A. - 85 : 12 (2014), p. 991-994. -
További szerzők:Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:OTKA Bridging Fund
OTKA
OSTRAT/810/213 University of Debrecen
Egyéb
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7.

001-es BibID:BIBFORM056762
Első szerző:Bene László (biofizikus)
Cím:The other side of the coin : time-domain fluorescence lifetime in flow / László Bene, László Damjanovich
Dátum:2015
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:Cytometry Part A. - 87 : 2 (2015), p. 101-103. -
További szerzők:Damjanovich László (1960-) (általános sebész)
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8.

001-es BibID:BIBFORM049378
Első szerző:Bene László (biofizikus)
Cím:Oxonol has the potential to probe membrane fields / László Bene
Dátum:2013
ISSN:1552-4922 1552-4930
Megjegyzések:MEMBRANE potential is an important physiological parameterof the living cell, which besides its traditional role in neurosciences,gains increasing attention also in the context of circulatingcells by recognizing its role in different transmembranesignalingprocesses such as immune-recognition (1), and in apoptosis(2,3). Besides its direct measurement with electrophysiologicaltechniques (electrodes and patch-clamp), indirecttechniques also exist. These are based on measuring fluorescenceof either the ionic dyes?such as the cationic 3,30-dihexyloxacarbocyanine(DiOC6(3)) and the anionic bis(1,3-dibutylbarbituricacid) trimethine oxonol (DiBAC4(3)) or bis-oxonol?responding with altered translocation or ratiometric dyes?suchas the merocyanine 540 (MC540), the 5,50,6,60-tetrachloro-1,10,3,30 tetraethylbenzimidazolocarbocyanine iodide (JC-1), the1-(3-sulfonatopropyl)24-{b[2-(di-n-octylamino)26-naphthyl]vinyl}pyridinium betaine (di-4-ANEPPS), and the 3-hydroxiflavones (e.g. F2N12S)?responding with spectralshifts (electrochromism) to the changes of membrane electricfields.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
absolute membrane potentia
calibration
flow cytometry
image cytometry
Nernstian dye
Megjelenés:Cytometry Part A. - 83A : 7 (2013), p. 608-611. -
Pályázati támogatás:TÁMOP-4.2.2.A-11/1/KONV-2012-0045
TÁMOP
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9.

001-es BibID:BIBFORM004939
Első szerző:Bene László (biofizikus)
Cím:Major histocompatibility complex class I protein conformation altered by transmembrane potential changes / Bene, L., Szollosi, J., Balazs, M., Matyus, L., Gaspar, R., Ameloot, M., Dale, R. E., Damjanovich, S.
Dátum:1997
ISSN:0196-4763
Megjegyzések:The nature of charge distributions in membrane-bound macromolecular structures renders them susceptible to interaction with transmembrane potential fields. As a result, conformational changes in such species may be expected to occur when this potential is altered. We have detected reversible conformational change in the major histocompatibility complex (MHC) class I antigen in the plasma membrane of human JY cells, as monitored by flow-cytometric resonance energy-transfer, upon reduction of the transmembrane potential (depolarization). This change increased the intramolecular energy-transfer efficiency between fluorescent donor- and acceptor-labeled monoclonal antibodies directed, respectively, to epitopes on the light (beta 2-microglobulin) and the heavy chains of the MHC class I antigen. Repolarization of the depolarized samples restored the energy-transfer efficiency to the original values measured before depolarization. Depolarization caused similar relative changes in fluorescence resonance energy-transfer efficiency when Fab fragments were used for labeling MHC class I complex, suggesting that the observed phenomenon is not restricted to whole monoclonal antibodies.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Antibodies, Monoclonal
Antigen Presentation
B-Lymphocytes
beta 2-Microglobulin
Cell Membrane
chemistry
cytology
Dyes
Energy Transfer
Enzyme Activation
Flow Cytometry
Fluorescein-5-isothiocyanate
Fluorescence
Fluorescent Dyes
Histocompatibility Antigens Class I
Human
Hungary
immunology
Light
Major Histocompatibility Complex
Membrane Potentials
metabolism
methods
Na(+)-K(+)-Exchanging ATPase
Patch-Clamp Techniques
physiology
Protein Conformation
Rhodamines
Surface Properties
Megjelenés:Cytometry. - 27 : 4 (1997), p. 353-357. -
További szerzők:Szöllősi János (1953-) (biofizikus) Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Mátyus László (1956-) (biofizikus) Gáspár Rezső (1944-) (biofizikus) Ameloot, Marcel Dale, Robert E. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:elektronikus változat
Borító:

10.

001-es BibID:BIBFORM004657
Első szerző:Bene László (biofizikus)
Cím:Detection of receptor clustering by flow cytometric fluorescence anisotropy measurements / Bene, L., Fulwyler, M. J., Damjanovich, S.
Dátum:2000
Megjegyzések:Perrin equation suggests an alternative way for the accurate energy transfer determination on a cell-by-cell basis by measuring polarized donor intensities in a conventional flow cytometer. METHODS: The relationship between energy transfer and fluorescence anisotropy of the donor was investigated by flow cytometric generation of Perrin-lifetime plots of fluorescent antibody-labeled MHC class I and class II molecules on the surface of living cells. The energy transfer reduced the fluorescence lifetime of the donor. RESULTS: Perrin plots have proven to be sensitive to the segmental mobility of the labeling dye and that of antibodies of different isotypes, and homo-transfer due to the multiple labeling of antibodies. A method demonstrating the feasibility of energy transfer determination by measuring anisotropy enhancement of the donor is presented. Flow cytometric histograms of the donor anisotropy and of the deduced energy transfer efficiency are shown, indicating clustering of MHC class I and class II molecules on the surface of human T lymphoblasts. In the Appendix, a method for the simultaneous determination of both energy transfer efficiency and donor fluorescence anisotropy, without need for G-factor measurement, is also presented. CONCLUSIONS: We demonstrate that energy transfer efficiency, i.e., proximity, between suitably selected donor and acceptor, and the rotational relaxation of the donor, i.e., donor mobility, can be simultaneously measured in a flow cytometer
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Adult
analysis
Antibodies
beta 2-Microglobulin
Cells
diagnostic use
Dyes
Energy Transfer
Flow Cytometry
Fluorescence
Fluorescence Polarization
Fluorescent Dyes
Histocompatibility Antigens Class I
Histocompatibility Antigens Class II
Human
Hungary
Immunoglobulin G
immunology
methods
Receptors,Cell Surface
Support,Non-U.S.Gov't
T-Lymphocytes
Megjelenés:Cytometry. - 40 : 4 (2000), p. 292-306. -
További szerzők:Fulwyler, Mack J. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:elektronikus változat
DOI
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11.

001-es BibID:BIBFORM030324
035-os BibID:(PMID)22128034
Első szerző:Damjanovich László (általános sebész)
Cím:Crohn's disease alters MHC-rafts in CD4+ T-cells / László Damjanovich, Julianna Volkó, Attila Forgács, Werner Hohenberger, László Bene
Dátum:2012
ISSN:1552-4922 1552-4930
Megjegyzések:Clusters of MHCI, ICAM-1, CD44, CD59, IL-2R, and IL-15R molecules have been studied on the surface of CD4(+) T-cells from peripheral blood and lymph nodes of patients in Crohn's disease and healthy individuals as controls by using a dual-laser flow cytometric fluorescence resonance energy transfer (FRET) technique and fluorescently stained Fabs. When cells from patients in Crohn's disease are compared to those of controls, the surface expression level for the MHCI reduced by ?45%, for CD44 enhanced by ?100%, and for IL-2R?, IL-15R?, and common ?(c) enhanced by ?50%, ?70%, and ?130%, respectively. Efficiencies of FRET monitoring homoassociation for the MHCI and CD44 reduced, that for IL-2R? enhanced. While efficiencies of FRET monitoring the association of ?(c) and ICAM-1 with the MHCI reduced, those monitoring association of IL-2/15R?, CD44, and CD59 with MHCI enhanced. Efficiencies of FRET measured between the MHCI and IL-2R?, IL-15R? differently enhanced to the advantage of IL-15R?, the one measured between ?(c) and IL-2R? reduced, suggesting modulations in the strength of interaction of MHCI with IL-2R, IL-15R, and ?(c) . The increases in density of surface bound cTx and in the associations of the receptors with the G(M1) -ganglioside lipid molecules suggest stronger lipid raft interactions of the receptors. The observed alterations of MHC-rafts in Crohn's disease-summarized in models of receptor patterns of diseased and control cells-may have functional consequences regarding signaling by the raft components.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Molekuláris Medicina
MHC class I
ICAM-1
IL-2R/15R
CD44
CD59
Crohn's disease
draining lymph node
CD4+ T-cells
lipid raft
fluorescence resonance energy transfer
Megjelenés:Cytometry. Part A. - 81A : 2 (2012), p. 149-164. -
További szerzők:Volkó Julianna (1983-) (biotechnológus) Forgács Attila (1985-) (fizikus) Hohenberger, Werner Bene László (1963-) (biofizikus)
Pályázati támogatás:K77600
OTKA
TÁMOP-4.2.1/B-09/1/KONV-2010-0007
TÁMOP
Sejt biofizika
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12.

001-es BibID:BIBFORM004878
035-os BibID:(scopus)26444586247 (wos)000232299300007
Első szerző:Nagy Péter (biofizikus)
Cím:Novel calibration method for flow cytometric fluorescence resonance energy transfer measurements between visible fluorescent proteins / Nagy, P., Bene, L., Hyun, W. C., Vereb, G., Braun, M., Antz, C., Paysan, J., Damjanovich, S., Park, J. W., Szollosi, J.
Dátum:2005
ISSN:1552-4922
Megjegyzések:The combination of fluorescence resonance energy transfer (FRET) and flow cytometry offers a statistically firm approach to study protein associations. Fusing green fluorescent protein (GFP) to a studied protein usually does not disturb the normal function of a protein, but quantitation of FRET efficiency calculated between GFP derivatives poses a problem in flow cytometry. METHODS: We generated chimeras in which cyan fluorescent protein (CFP) was separated by amino acid linkers of different sizes from yellow fluorescent protein (YFP) and used them to calibrate the cell-by-cell flow cytometric FRET measurements carried out on two different dual-laser flow cytometers. Then, CFP-Kip1 was coexpressed in yeast cells with YFP and cyclin-dependent kinase-2 (Cdk2) and served as a positive control for FRET measurements, and CFP-Kip1 coexpressed with a random peptide fused to YFP was the negative control. RESULTS: We measured donor, direct, and sensitized acceptor fluorescence intensities and developed a novel way to calculate a factor (alpha) that characterized the fluorescence intensity of acceptor molecules relative to the same number of excited donor molecules, which is essential for quantifying FRET efficiency. This was achieved by calculating FRET efficiency in two different ways and minimizing the squared difference between the two results by changing alpha. Our method reliably detected the association of Cdk2 with its inhibitor, Kip1, whereas the nonspecific FRET efficiency between Cdk2 and a random peptide was negligible. We identified and sorted subpopulations of yeast cells showing interaction between the studied proteins. CONCLUSIONS: We have described a straightforward novel calibration method to accurately quantitate FRET efficiency between GFP derivatives in flow cytometry
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
analysis
Biophysics
Calibration
Cells
chemistry
Cyclin-Dependent Kinase 2
Cyclin-Dependent Kinase Inhibitor p27
Energy Transfer
Flow Cytometry
Fluorescence
Fluorescence Resonance Energy Transfer
Hungary
metabolism
methods
Protein Binding
Proteins
Research
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Support
Megjelenés:Cytometry. Part A. - 67 : 2 (2005), p. 86-96. -
További szerzők:Bene László (1963-) (biofizikus) Hyun, William C. Vereb György (1965-) (biofizikus, orvos) Braun, Manuel Antz, Christof Paysan, Jacques Damjanovich Sándor (1936-2017) (biofizikus) Park, John W. Szöllősi János (1953-) (biofizikus)
Internet cím:elektronikus változat
DOI
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