Log in

Relevant bibliographies by topics / FLIM-FRET / Journal articles

To see the other types of publications on this topic, follow the link: FLIM-FRET.

Journal articles on the topic 'FLIM-FRET'

Author: Grafiati

Published: 4 June 2021

Last updated: 27 April 2022

Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles

Select a source type:

Consult the top 50 journal articles for your research on the topic 'FLIM-FRET.'

Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.

You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.

Browse journal articles on a wide variety of disciplines and organise your bibliography correctly.

1

Garsha, Karl. "A Comment on using FLIM with FRET." Microscopy Today 14, no. 3 (May 2006): 52–53. http://dx.doi.org/10.1017/s1551929500057709.

Full text

Abstract:

Depending on the nature of the study and what sort of information one is trying to gather through the use of FRET, FLIM has some compelling advantages in certain situations, and can provide a quantitative evaluation of the donor, acceptor and FRET pair stoichiometry. It does require access to specialized equipment and software. Different approaches to FLIM data acquisition have different strengths and weaknesses. For dynamic studies requiring high time resolution, FLIM acquisition times can fall well short of ideal.If a yes/no answer to whether FRET is occurring is all that is required, then the polarization anisotropy of the acceptor can be used to determine FRET between fluorescent proteins (Rizzo and Piston, 2005). This is a relatively simple and robust method for confirming the presence/absence of FRET.

APA, Harvard, Vancouver, ISO, and other styles

2

Lee, Jiung-De, Ping-Chun Huang, Yi-Cheng Lin, Lung-Sen Kao, Chien-Chang Huang, Fu-Jen Kao, Chung-Chih Lin, and De-Ming Yang. "In-Depth Fluorescence Lifetime Imaging Analysis Revealing SNAP25A-Rabphilin 3A Interactions." Microscopy and Microanalysis 14, no. 6 (November 6, 2008): 507–18. http://dx.doi.org/10.1017/s1431927608080628.

Full text

Abstract:

AbstractThe high sensitivity and spatial resolution enabled by two-photon excitation fluorescence lifetime imaging microscopy/fluorescence resonance energy transfer (2PE-FLIM/FRET) provide an effective approach that reveals protein-protein interactions in a single cell during stimulated exocytosis. Enhanced green fluorescence protein (EGFP)–labeled synaptosomal associated protein of 25 kDa (SNAP25A) and red fluorescence protein (mRFP)–labeled Rabphillin 3A (RPH3A) were co-expressed in PC12 cells as the FRET donor and acceptor, respectively. The FLIM images of EGFP-SNAP25A suggested that SNAP25A/RPH3A interaction was increased during exocytosis. In addition, the multidimensional (three-dimensional with time) nature of the 2PE-FLIM image datasets can also resolve the protein interactions in the z direction, and we have compared several image analysis methods to extract more accurate and detailed information from the FLIM images. Fluorescence lifetime was fitted by using one and two component analysis. The lifetime FRET efficiency was calculated by the peak lifetime (τpeak) and the left side of the half-peak width (τ1/2), respectively. The results show that FRET efficiency increased at cell surface, which suggests that SNAP25A/RPH3A interactions take place at cell surface during stimulated exocytosis. In summary, we have demonstrated that the 2PE-FLIM/FRET technique is a powerful tool to reveal dynamic SNAP25A/RPH3A interactions in single neuroendocrine cells.

APA, Harvard, Vancouver, ISO, and other styles

3

Rajoria, Shilpi, Lingling Zhao, Xavier Intes, and Margarida Barroso. "FLIM-FRET for Cancer Applications." Current Molecular Imaging 3, no. 2 (February 4, 2015): 144–61. http://dx.doi.org/10.2174/2211555203666141117221111.

Full text

APA, Harvard, Vancouver, ISO, and other styles

4

Bücherl, Christoph A., Arjen Bader, Adrie H. Westphal, Sergey P. Laptenok, and Jan Willem Borst. "FRET-FLIM applications in plant systems." Protoplasma 251, no. 2 (January 4, 2014): 383–94. http://dx.doi.org/10.1007/s00709-013-0595-7.

Full text

APA, Harvard, Vancouver, ISO, and other styles

5

Wang, Shiqi, Binglin Shen, Sheng Ren, Yihua Zhao, Silu Zhang, Junle Qu, and Liwei Liu. "Implementation and application of FRET–FLIM technology." Journal of Innovative Optical Health Sciences 12, no. 05 (September 2019): 1930010. http://dx.doi.org/10.1142/s1793545819300106.

Full text

Abstract:

With the development of the new detection methods and the function of fluorescent molecule, researchers hope to further explore the internal mechanisms of organisms, monitor changes in the intracellular microenvironment, and dynamic processes of molecular interactions in cells. Fluorescence resonance energy transfer (FRET) describes the energy transfer process between donor fluorescent molecules and acceptor fluorescent molecules. It is an important means to detect protein–protein interactions and protein conformation changes in cells. Fluorescence lifetime imaging microscopy (FLIM) enables noninvasive measurement of the fluorescence lifetime of fluorescent particles in vivo. The FRET–FLIM technology, which is use FLIM to quantify and analyze FRET, enables real-time monitoring of dynamic changes of proteins in biological cells and analysis of protein interaction mechanisms. The distance between donor and acceptor and their respective fluorescent lifetime, which are of great importance for studying the mechanism of intracellular activity can be obtained by data analysis and algorithm fitting.

APA, Harvard, Vancouver, ISO, and other styles

6

Ellis, Jonathan D., David Llères, Marco Denegri, Angus I. Lamond, and Javier F. Cáceres. "Spatial mapping of splicing factor complexes involved in exon and intron definition." Journal of Cell Biology 181, no. 6 (June 16, 2008): 921–34. http://dx.doi.org/10.1083/jcb.200710051.

Full text

Abstract:

We have analyzed the interaction between serine/arginine-rich (SR) proteins and splicing components that recognize either the 5′ or 3′ splice site. Previously, these interactions have been extensively characterized biochemically and are critical for both intron and exon definition. We use fluorescence resonance energy transfer (FRET) microscopy to identify interactions of individual SR proteins with the U1 small nuclear ribonucleoprotein (snRNP)–associated 70-kD protein (U1 70K) and with the small subunit of the U2 snRNP auxiliary factor (U2AF35) in live-cell nuclei. We find that these interactions occur in the presence of RNA polymerase II inhibitors, demonstrating that they are not exclusively cotranscriptional. Using FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM), we map these interactions to specific sites in the nucleus. The FLIM data also reveal a previously unknown interaction between HCC1, a factor related to U2AF65, with both subunits of U2AF. Spatial mapping using FLIM-FRET reveals differences in splicing factors interactions within complexes located in separate subnuclear domains.

APA, Harvard, Vancouver, ISO, and other styles

7

Llères, David, John James, Sam Swift, David G. Norman, and Angus I. Lamond. "Quantitative analysis of chromatin compaction in living cells using FLIM–FRET." Journal of Cell Biology 187, no. 4 (November 16, 2009): 481–96. http://dx.doi.org/10.1083/jcb.200907029.

Full text

Abstract:

We present a quantitative Förster resonance energy transfer (FRET)–based assay using multiphoton fluorescence lifetime imaging microscopy (FLIM) to measure chromatin compaction at the scale of nucleosomal arrays in live cells. The assay uses a human cell line coexpressing histone H2B tagged to either enhanced green fluorescent protein (FP) or mCherry FPs (HeLaH2B-2FP). FRET occurs between FP-tagged histones on separate nucleosomes and is increased when chromatin compacts. Interphase cells consistently show three populations of chromatin with low, medium, or high FRET efficiency, reflecting spatially distinct regions with different levels of chromatin compaction. Treatment with inhibitors that either increase chromatin compaction (i.e., depletion of adenosine triphosphate) or decrease chromosome compaction (trichostatin A) results in a parallel increase or decrease in the FLIM–FRET signal. In mitosis, the assay showed variation in compaction level, as reflected by different FRET efficiency populations, throughout the length of all chromosomes, increasing to a maximum in late anaphase. These data are consistent with extensive higher order folding of chromatin fibers taking place during anaphase.

APA, Harvard, Vancouver, ISO, and other styles

8

Kelly, Douglas J., Sean C. Warren, Dominic Alibhai, Sunil Kumar, Yuriy Alexandrov, Ian Munro, Anca Margineanu, et al. "Automated multiwell fluorescence lifetime imaging for Förster resonance energy transfer assays and high content analysis." Analytical Methods 7, no. 10 (2015): 4071–89. http://dx.doi.org/10.1039/c5ay00244c.

Full text

APA, Harvard, Vancouver, ISO, and other styles

9

Sambrook, Joseph, and David W. Russell. "Probing Protein Interactions Using GFP and FRET Stage 3: FLIM-FRET Measurements." Cold Spring Harbor Protocols 2006, no. 1 (June 2006): pdb.prot3822. http://dx.doi.org/10.1101/pdb.prot3822.

Full text

APA, Harvard, Vancouver, ISO, and other styles

10

Kelleher, M. T., F. Festy, P. R. Barber, C. Gillett, E. Ofo, A. Coolen, S. Pinder, et al. "Use of novel optical proteomics to profile breast cancer patients leading to individualised prognosis and tailored treatment." Journal of Clinical Oncology 27, no. 15_suppl (May 20, 2009): e22090-e22090. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.e22090.

Full text

Abstract:

e22090 Background: Optical proteomics quantifies interactions between proteins and post-translational modifications by measuring Förster resonance energy transfer (FRET) quantified by fluorescence lifetime imaging microscopy (FLIM). This project aims to derive multiple high throughput optical proteomic markers, to predict metastatic risk at first diagnosis, and to perturb ‘high risk' protein-protein interactions using targeted therapeutics. This initial step develops robust FRET/FLIM assays, suitable for use in formalin fixed paraffin embedded (FFPE) tissue to be correlated with patient outcome. Methods: Fluorophore-conjugated antibodies to proteins involved in cell migration and survival, were applied to tissue microarrays (TMA), created from archived FFPE invasive ductal breast carcinoma samples. Where fluorophores are located within nanometer proximity, FRET occurs, thus allowing quantification of protein-protein interaction. Ezrin and PKCα phosphorylation, distribution, and interaction were imaged on four TMAs (patients diagnosed with early breast cancer 1984 -1987: 20 years follow-up data). Results: 71 patient samples were optically imaged. Patients were clustered based on the pairwise distances between 18 optical variables ‘input data'. Data are represented on self organising maps and dendrograms and correlated with clinical outcome ‘output data', displaying a heatmap distribution. Conclusions: Ezrin and PKCα phosphorylation, distribution, and interaction imaged optically within FFPE contain prognostic information regarding metastatic outcome in breast cancer, thus stepping ever closer to individualising prognosis. These advanced optics-based parameters informing on metastatic potential will be validated in prospective studies in conjunction with FRET/FLIM assays measuring HER2/HER3 dimerisation, and EGFR and HER2 ubiquitination in order to improve patient selection for targeted therapy. No significant financial relationships to disclose.

APA, Harvard, Vancouver, ISO, and other styles

11

Verveer, P. J., O. Rocks, A. G. Harpur, and P. I. H. Bastiaens. "Imaging Protein Interactions by FRET Microscopy: FLIM Measurements." Cold Spring Harbor Protocols 2006, no. 6 (November 1, 2006): pdb.prot4599. http://dx.doi.org/10.1101/pdb.prot4599.

Full text

APA, Harvard, Vancouver, ISO, and other styles

12

Wallrabe, Horst, and Ammasi Periasamy. "Imaging protein molecules using FRET and FLIM microscopy." Current Opinion in Biotechnology 16, no. 1 (February 2005): 19–27. http://dx.doi.org/10.1016/j.copbio.2004.12.002.

Full text

APA, Harvard, Vancouver, ISO, and other styles

13

Kaufmann, Tanja, Sébastien Herbert, Benjamin Hackl, Johanna Maria Besold, Christopher Schramek, Josef Gotzmann, Kareem Elsayad, and Dea Slade. "Direct measurement of protein–protein interactions by FLIM-FRET at UV laser-induced DNA damage sites in living cells." Nucleic Acids Research 48, no. 21 (October 14, 2020): e122-e122. http://dx.doi.org/10.1093/nar/gkaa859.

Full text

Abstract:

Abstract Protein–protein interactions are essential to ensure timely and precise recruitment of chromatin remodellers and repair factors to DNA damage sites. Conventional analyses of protein–protein interactions at a population level may mask the complexity of interaction dynamics, highlighting the need for a method that enables quantification of DNA damage-dependent interactions at a single-cell level. To this end, we integrated a pulsed UV laser on a confocal fluorescence lifetime imaging (FLIM) microscope to induce localized DNA damage. To quantify protein–protein interactions in live cells, we measured Förster resonance energy transfer (FRET) between mEGFP- and mCherry-tagged proteins, based on the fluorescence lifetime reduction of the mEGFP donor protein. The UV-FLIM-FRET system offers a unique combination of real-time and single-cell quantification of DNA damage-dependent interactions, and can distinguish between direct protein–protein interactions, as opposed to those mediated by chromatin proximity. Using the UV-FLIM-FRET system, we show the dynamic changes in the interaction between poly(ADP-ribose) polymerase 1, amplified in liver cancer 1, X-ray repair cross-complementing protein 1 and tripartite motif containing 33 after DNA damage. This new set-up complements the toolset for studying DNA damage response by providing single-cell quantitative and dynamic information about protein–protein interactions at DNA damage sites.

APA, Harvard, Vancouver, ISO, and other styles

14

Huang, Ping-Chun, Tai-Yu Chiu, Li-Chun Wang, Hsiao-Chuan Teng, Fu-Jen Kao, and De-Ming Yang. "Visualization of the Orai1 Homodimer and the Functional Coupling of Orai1-STIM1 by Live-Cell Fluorescence Lifetime Imaging." Microscopy and Microanalysis 16, no. 3 (April 9, 2010): 313–26. http://dx.doi.org/10.1017/s1431927610000188.

Full text

Abstract:

AbstractThe Orai1-STIM1 constructed store-operated Ca2+ channels (SOCs) have been found to exert several essential Ca2+ entry/signaling cascades, e.g., the generation of immune response in T lymphocytes. Although biochemical and novel imaging evidence appear to indicate that Orai1 and STIM1 interact with each other to achieve store-operated Ca2+ entry (SOCE), the detailed mechanism of functional SOCE in situ has yet to be fully understood. In this study, green fluorescence protein (EGFP as donor) targeted to either the N- or C-terminal of Orai1 (wild type or ▵1-90+▵267-301 double deletion type) and mOrange (as acceptor) tagged STIM1 were used to comprise a fluorescence resonance energy transfer (FRET) pair within living PC12 cells. The fluorescence lifetime map and histogram/distribution of each single cell, determined by one-photon excitation fluorescence lifetime imaging microscopy (FLIM), was used to visualize FRET and show the Orai1 homodimer and Orai1-STIM1 binding. Both the color-coded lifetime map and the distribution of EGFP-tagged Orai1 significantly changed after the administration of thapsigargin, the SOCE stimulating agent. The FRET efficiency from each experimental set was also calculated and compared using double exponential analysis. In summary, we show the detailed interactions Orai1-Orai1 and Orai1-STIM1 within intact living cells by using the FLIM-FRET technique.

APA, Harvard, Vancouver, ISO, and other styles

15

Nair, Deepak K., Mini Jose, Thomas Kuner, Werner Zuschratter, and Roland Hartig. "FRET-FLIM at nanometer spectral resolution from living cells." Optics Express 14, no. 25 (2006): 12217. http://dx.doi.org/10.1364/oe.14.012217.

Full text

APA, Harvard, Vancouver, ISO, and other styles

16

Shcherbo, Dmitry, Ekaterina A. Souslova, Joachim Goedhart, Tatyana V. Chepurnykh, Anna Gaintzeva, Irina I. Shemiakina, Theodorus WJ Gadella, Sergey Lukyanov, and Dmitriy M. Chudakov. "Practical and reliable FRET/FLIM pair of fluorescent proteins." BMC Biotechnology 9, no. 1 (2009): 24. http://dx.doi.org/10.1186/1472-6750-9-24.

Full text

APA, Harvard, Vancouver, ISO, and other styles

17

Radić, Martina, Marko Šoštar, Igor Weber, Helena Ćetković, Neda Slade, and Maja Herak Bosnar. "The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage." International Journal of Molecular Sciences 21, no. 7 (March 29, 2020): 2363. http://dx.doi.org/10.3390/ijms21072363.

Full text

Abstract:

Nucleoside diphosphate kinases (NDPK/NME/Nm23) are enzymes composed of subunits NME1/NDPK A and NME2/NDPK B, responsible for the maintenance of the cellular (d)NTP pool and involved in other cellular processes, such as metastasis suppression and DNA damage repair. Although eukaryotic NDPKs are active only as hexamers, it is unclear whether other NME functions require the hexameric form, and how the isoenzyme composition varies in different cellular compartments. To examine the effect of DNA damage on intracellular localization of NME1 and NME2 and the composition of NME oligomers in the nucleus and the cytoplasm, we used live-cell imaging and the FRET/FLIM technique. We showed that exogenous NME1 and NME2 proteins co-localize in the cytoplasm of non-irradiated cells, and move simultaneously to the nucleus after gamma irradiation. The FRET/FLIM experiments imply that, after DNA damage, there is a slight shift in the homomer/heteromer balance between the nucleus and the cytoplasm. Collectively, our results indicate that, after irradiation, NME1 and NME2 engage in mutual functions in the nucleus, possibly performing specific functions in their homomeric states. Finally, we demonstrated that fluorophores fused to the N-termini of NME polypeptides produce the largest FRET effect and thus recommend this orientation for use in similar studies.

APA, Harvard, Vancouver, ISO, and other styles

18

Rainey, Kristin H., and George H. Patterson. "Photoswitching FRET to monitor protein–protein interactions." Proceedings of the National Academy of Sciences 116, no. 3 (December 31, 2018): 864–73. http://dx.doi.org/10.1073/pnas.1805333116.

Full text

Abstract:

FRET is a powerful approach to study the interactions of fluorescent molecules, and numerous methods have been developed to measure FRET in cells. Here, we present a method based on a donor molecule’s photoswitching properties, which are slower in the presence vs. the absence of an acceptor. The technique, photoswitching FRET (psFRET), is similar to an established but underutilized method called photobleaching FRET (pbFRET), with the major difference being that the molecules are switched “off” rather than photobleached. The psFRET technique has some of the FRET imaging advantages normally attributed to fluorescence lifetime imaging microscopy (FLIM), such as monitoring only donor fluorescence. However, it can be performed on a conventional widefield microscope, requires less illumination light to photoswitch off than photobleaching, and can be photoswitched “on” again to repeat the experiment. We present data testing the validity of the psFRET approach to quantify FRET in cells and demonstrate its use in imaging protein–protein interactions and fluorescent protein-based biosensors.

APA, Harvard, Vancouver, ISO, and other styles

19

Duan, Zhikun, Kaiwen Li, Wenwen Duan, Junli Zhang, and Jingjing Xing. "Probing membrane protein interactions and signaling molecule homeostasis in plants by Förster resonance energy transfer analysis." Journal of Experimental Botany 73, no. 1 (October 5, 2021): 68–77. http://dx.doi.org/10.1093/jxb/erab445.

Full text

Abstract:

Abstract Membrane proteins have key functions in signal transduction, transport, and metabolism. Therefore, deciphering the interactions between membrane proteins provides crucial information on signal transduction and the spatiotemporal organization of protein complexes. However, detecting the interactions and behaviors of membrane proteins in their native environments remains difficult. Förster resonance energy transfer (FRET) is a powerful tool for quantifying the dynamic interactions and assembly of membrane proteins without disrupting their local environment, supplying nanometer-scale spatial information and nanosecond-scale temporal information. In this review, we briefly introduce the basic principles of FRET and assess the current state of progress in the development of new FRET techniques (such as FRET-FLIM, homo-FRET, and smFRET) for the analysis of plant membrane proteins. We also describe the various FRET-based biosensors used to quantify the homeostasis of signaling molecules and the active state of kinases. Furthermore, we summarize recent applications of these advanced FRET sensors in probing membrane protein interactions, stoichiometry, and protein clustering, which have shed light on the complex biological functions of membrane proteins in living plant cells.

APA, Harvard, Vancouver, ISO, and other styles

20

Sambrook, Joseph, and David W. Russell. "Probing Protein Interactions Using GFP and FRET Stage 2: Cell Preparation for FLIM-FRET Analysis." Cold Spring Harbor Protocols 2006, no. 1 (June 2006): pdb.prot3893. http://dx.doi.org/10.1101/pdb.prot3893.

Full text

APA, Harvard, Vancouver, ISO, and other styles

21

Ishikawa-Ankerhold, Hellen C., Richard Ankerhold, and Gregor P. C. Drummen. "Advanced Fluorescence Microscopy Techniques—FRAP, FLIP, FLAP, FRET and FLIM." Molecules 17, no. 4 (April 2, 2012): 4047–132. http://dx.doi.org/10.3390/molecules17044047.

Full text

APA, Harvard, Vancouver, ISO, and other styles

22

Heinrich, Philippe, Mariano Gonzalez Pisfil, Jonas Kahn, Laurent Héliot, and Aymeric Leray. "Implementation of Transportation Distance for Analyzing FLIM and FRET Experiments." Bulletin of Mathematical Biology 76, no. 10 (September 25, 2014): 2596–626. http://dx.doi.org/10.1007/s11538-014-0025-9.

Full text

APA, Harvard, Vancouver, ISO, and other styles

23

Adbul Rahim, Nur Aida, Serge Pelet, Roger D. Kamm, and Peter T. C. So. "Methodological considerations for global analysis of cellular FLIM/FRET measurements." Journal of Biomedical Optics 17, no. 2 (2012): 026013. http://dx.doi.org/10.1117/1.jbo.17.2.026013.

Full text

APA, Harvard, Vancouver, ISO, and other styles

24

Hallworth, Richard, Benjamin Currall, Michael G. Nichols, Xudong Wu, and Jian Zuo. "Studying inner ear protein–protein interactions using FRET and FLIM." Brain Research 1091, no. 1 (May 2006): 122–31. http://dx.doi.org/10.1016/j.brainres.2006.02.076.

Full text

APA, Harvard, Vancouver, ISO, and other styles

25

Mountford, C. P., A. R. Mount, S. A. G. Evans, T. J. Su, P. Dickinson, A. H. Buck, C. J. Campbell, et al. "Time-Resolved FRET and FLIM of Four-way DNA Junctions." Journal of Fluorescence 16, no. 6 (September 22, 2006): 839–45. http://dx.doi.org/10.1007/s10895-006-0125-5.

Full text

APA, Harvard, Vancouver, ISO, and other styles

26

Periasamy, A., and Y. Sun. "Monitoring Protein-Protein Interactions in Living Specimens using FLIM-FRET Microscopy." Microscopy and Microanalysis 18, S2 (July 2012): 148–49. http://dx.doi.org/10.1017/s1431927612002590.

Full text

APA, Harvard, Vancouver, ISO, and other styles

27

Chen, Ye, James D. Mills, and Ammasi Periasamy. "Protein localization in living cells and tissues using FRET and FLIM." Differentiation 71, no. 9-10 (December 2003): 528–41. http://dx.doi.org/10.1111/j.1432-0436.2003.07109007.x.

Full text

APA, Harvard, Vancouver, ISO, and other styles

28

Grecco, Hernan E., Pedro Roda-Navarro, and Peter J. Verveer. "Global analysis of time correlated single photon counting FRET-FLIM data." Optics Express 17, no. 8 (April 3, 2009): 6493. http://dx.doi.org/10.1364/oe.17.006493.

Full text

APA, Harvard, Vancouver, ISO, and other styles

29

Sauer, Benjamin, Qinghai Tian, Peter Lipp, and Lars Kaestner. "Confocal FLIM of Genetically Encoded FRET Sensors for Quantitative Ca2+Imaging." Cold Spring Harbor Protocols 2014, no. 12 (December 2014): pdb.prot077040. http://dx.doi.org/10.1101/pdb.prot077040.

Full text

APA, Harvard, Vancouver, ISO, and other styles

30

Pelicci, Simone, Michele Oneto, Melody Di Bona, Isotta Cainero, Paola Barboro, Alberto Diaspro, and Luca Lanzano′. "Chromatin Nanoscale Organization Investigated by FLIM-FRET and STED Superresolution Microscopy." Biophysical Journal 116, no. 3 (February 2019): 174a. http://dx.doi.org/10.1016/j.bpj.2018.11.965.

Full text

APA, Harvard, Vancouver, ISO, and other styles

31

Quan, My D., Shih-Chu Liao, Josephine C. Ferreon, and Allan Chris M. Ferreon. "Protein conformations in Tau condensates probed by smFRET and FLIM-FRET." Biophysical Journal 121, no. 3 (February 2022): 472a. http://dx.doi.org/10.1016/j.bpj.2021.11.412.

Full text

APA, Harvard, Vancouver, ISO, and other styles

32

Thaa, Bastian, Andreas Herrmann, and Michael Veit. "Intrinsic Cytoskeleton-Dependent Clustering of Influenza Virus M2 Protein with Hemagglutinin Assessed by FLIM-FRET." Journal of Virology 84, no. 23 (September 29, 2010): 12445–49. http://dx.doi.org/10.1128/jvi.01322-10.

Full text

Abstract:

ABSTRACT The hemagglutinin (HA) of influenza virus organizes the virus bud zone, a domain of the plasma membrane enriched in raft lipids. Using fluorescence lifetime imaging microscopy-fluorescence resonance energy transfer (FLIM-FRET), a technique that detects close colocalization of fluorescent proteins in transfected cells, we show that the viral proton channel M2 clusters with HA but not with a marker for inner leaflet rafts. The FRET signal between M2 and HA depends on the raft-targeting signals in HA and on an intact actin cytoskeleton. We conclude that M2 contains an intrinsic signal that targets the protein to the viral bud zone, which is organized by raft-associated HA and by cortical actin.

APA, Harvard, Vancouver, ISO, and other styles

33

Kaye, Bryan, Tae Yeon Yoo, Peter J. Foster, Che-Hang Yu, and Daniel J. Needleman. "Bridging length scales to measure polymer assembly." Molecular Biology of the Cell 28, no. 10 (May 15, 2017): 1379–88. http://dx.doi.org/10.1091/mbc.e16-05-0344.

Full text

Abstract:

Time-resolvable quantitative measurements of polymer concentration are very useful to elucidate protein polymerization pathways. There are numerous techniques to measure polymer concentrations in purified protein solutions, but few are applicable in vivo. Here we develop a methodology combining microscopy and spectroscopy to overcome the limitations of both approaches for measuring polymer concentration in cells and cell extracts. This technique is based on quantifying the relationship between microscopy and spectroscopy measurements at many locations. We apply this methodology to measure microtubule assembly in tissue culture cells and Xenopus egg extracts using two-photon microscopy with FLIM measurements of FRET. We find that the relationship between FRET and two-photon intensity quantitatively agrees with predictions. Furthermore, FRET and intensity measurements change as expected with changes in acquisition time, labeling ratios, and polymer concentration. Taken together, these results demonstrate that this approach can quantitatively measure microtubule assembly in complex environments. This methodology should be broadly useful for studying microtubule nucleation and assembly pathways of other polymers.

APA, Harvard, Vancouver, ISO, and other styles

34

Oliveira, Ana F., and Ryohei Yasuda. "An Improved Ras Sensor for Highly Sensitive and Quantitative FRET-FLIM Imaging." PLoS ONE 8, no. 1 (January 14, 2013): e52874. http://dx.doi.org/10.1371/journal.pone.0052874.

Full text

APA, Harvard, Vancouver, ISO, and other styles

35

Padilla-Parra, Sergi, Nicolas Audugé, Hervé Lalucque, Jean-Claude Mevel, Maïté Coppey-Moisan, and Marc Tramier. "Quantitative Comparison of Different Fluorescent Protein Couples for Fast FRET-FLIM Acquisition." Biophysical Journal 97, no. 8 (October 2009): 2368–76. http://dx.doi.org/10.1016/j.bpj.2009.07.044.

Full text

APA, Harvard, Vancouver, ISO, and other styles

36

Demeautis, Claire, François Sipieter, Julien Roul, Catherine Chapuis, Sergi Padilla-Parra, Franck Riquet, and Marc Tramier. "Single Wavelength Excitation Dual Color Flim for Multiplexing Genetically Encoded FRET Biosensors." Biophysical Journal 110, no. 3 (February 2016): 518a. http://dx.doi.org/10.1016/j.bpj.2015.11.2771.

Full text

APA, Harvard, Vancouver, ISO, and other styles

37

Altenbach, Kirsten, Rory R. Duncan, and Mari Valkonen. "In vivo FLIM-FRET measurements of recombinant proteins expressed in filamentous fungi." Fungal Biology Reviews 23, no. 3 (August 2009): 67–71. http://dx.doi.org/10.1016/j.fbr.2009.12.002.

Full text

APA, Harvard, Vancouver, ISO, and other styles

38

Pelicci, Simone, Michele Oneto, Melody Di Bona, Alberto Diaspro, and Luca Lanzanò. "FLIM-FRET of Chromatin in Live Cells using Two DNA-Binding Dyes." Biophysical Journal 114, no. 3 (February 2018): 532a. http://dx.doi.org/10.1016/j.bpj.2017.11.2909.

Full text

APA, Harvard, Vancouver, ISO, and other styles

39

Guo, Wenjun, Sunil Kumar, Frederik Görlitz, Edwin Garcia, Yuriy Alexandrov, Ian Munro, Douglas J. Kelly, et al. "Automated Fluorescence Lifetime Imaging High-Content Analysis of Förster Resonance Energy Transfer between Endogenously Labeled Kinetochore Proteins in Live Budding Yeast Cells." SLAS TECHNOLOGY: Translating Life Sciences Innovation 24, no. 3 (January 10, 2019): 308–20. http://dx.doi.org/10.1177/2472630318819240.

Full text

Abstract:

We describe an open-source automated multiwell plate fluorescence lifetime imaging (FLIM) methodology to read out Förster resonance energy transfer (FRET) between fluorescent proteins (FPs) labeling endogenous kinetochore proteins (KPs) in live budding yeast cells. The low copy number of many KPs and their small spatial extent present significant challenges for the quantification of donor fluorescence lifetime in the presence of significant cellular autofluorescence and photobleaching. Automated FLIM data acquisition was controlled by µManager and incorporated wide-field time-gated imaging with optical sectioning to reduce background fluorescence. For data analysis, we used custom MATLAB-based software tools to perform kinetochore foci segmentation and local cellular background subtraction and fitted the fluorescence lifetime data using the open-source FLIMfit software. We validated the methodology using endogenous KPs labeled with mTurquoise2 FP and/or yellow FP and measured the donor fluorescence lifetimes for foci comprising 32 kinetochores with KP copy numbers as low as ~2 per kinetochore under an average labeling efficiency of 50%. We observed changes of median donor lifetime ≥250 ps for KPs known to form dimers. Thus, this FLIM high-content analysis platform enables the screening of relatively low-copy-number endogenous protein–protein interactions at spatially confined macromolecular complexes.

APA, Harvard, Vancouver, ISO, and other styles

40

Carmichael, Stephen W. "Looking at the Functional State of Proteins Inside Cells." Microscopy Today 7, no. 8 (October 1999): 3–4. http://dx.doi.org/10.1017/s155192950006497x.

Full text

Abstract:

Many intracellular proteins are catalysts that regulate cellular functions. These catalysts can be assayed to determine their functional state, but untii now it was not possible to simultaneously obtain a functional analysis and spatial data. Tony Ng, Anthony Squire, and others, working in the laboratories of Phillippe Bastiaens and Peter Parker, have combined Fluorescence Lifetime Imaging Microscopy (FLIM) with Fluorescence Resonance Energy Transfer (FRET) to spatially resolve the activation of a protein catalyst within living cells. Their technique was also applied to fixed cells.

APA, Harvard, Vancouver, ISO, and other styles

41

Carlon-Andres, Irene, and Sergi Padilla-Parra. "Quantitative FRET-FLIM-BlaM to Assess the Extent of HIV-1 Fusion in Live Cells." Viruses 12, no. 2 (February 12, 2020): 206. http://dx.doi.org/10.3390/v12020206.

Full text

Abstract:

The first steps of human immunodeficiency virus (HIV) infection go through the engagement of HIV envelope (Env) with CD4 and coreceptors (CXCR4 or CCR5) to mediate viral membrane fusion between the virus and the host. New approaches are still needed to better define both the molecular mechanistic underpinnings of this process but also the point of fusion and its kinetics. Here, we have developed a new method able to detect and quantify HIV-1 fusion in single live cells. We present a new approach that employs fluorescence lifetime imaging microscopy (FLIM) to detect Förster resonance energy transfer (FRET) when using the β-lactamase (BlaM) assay. This novel approach allows comparing different populations of single cells regardless the concentration of CCF2-AM FRET reporter in each cell, and more importantly, is able to determine the relative amount of viruses internalized per cell. We have applied this approach in both reporter TZM-bl cells and primary T cell lymphocytes.

APA, Harvard, Vancouver, ISO, and other styles

42

Xu, Lingling, Liang Wang, Zhihong Zhang, and Zhen-Li Huang. "A Feasible Add-On Upgrade on a Commercial Two-Photon FLIM Microscope for Optimal FLIM-FRET Imaging of CFP-YFP Pairs." Journal of Fluorescence 23, no. 3 (March 3, 2013): 543–49. http://dx.doi.org/10.1007/s10895-013-1188-8.

Full text

APA, Harvard, Vancouver, ISO, and other styles

43

Omer, Travis, Xavier Intes, and Juergen Hahn. "Temporal Data Set Reduction Based on D-Optimality for Quantitative FLIM-FRET Imaging." PLOS ONE 10, no. 12 (December 11, 2015): e0144421. http://dx.doi.org/10.1371/journal.pone.0144421.

Full text

APA, Harvard, Vancouver, ISO, and other styles

44

Engel, Stephanie, Silvia Scolari, Bastian Thaa, Nils Krebs, Thomas Korte, Andreas Herrmann, and Michael Veit. "FLIM-FRET and FRAP reveal association of influenza virus haemagglutinin with membrane rafts." Biochemical Journal 425, no. 3 (January 15, 2010): 567–73. http://dx.doi.org/10.1042/bj20091388.

Full text

Abstract:

It has been supposed that the HA (haemagglutinin) of influenza virus must be recruited to membrane rafts to perform its function in membrane fusion and virus budding. In the present study, we aimed at substantiating this association in living cells by biophysical methods. To this end, we fused the cyan fluorescent protein Cer (Cerulean) to the cytoplasmic tail of HA. Upon expression in CHO (Chinese-hamster ovary) cells HA–Cer was glycosylated and transported to the plasma membrane in a similar manner to authentic HA. We measured FLIM-FRET (Förster resonance energy transfer by fluorescence lifetime imaging microscopy) and showed strong association of HA–Cer with Myr-Pal–YFP (myristoylated and palmitoylated peptide fused to yellow fluorescent protein), an established marker for rafts of the inner leaflet of the plasma membrane. Clustering was significantly reduced when rafts were disintegrated by cholesterol extraction and when the known raft-targeting signals of HA, the palmitoylation sites and amino acids in its transmembrane region, were removed. FRAP (fluorescence recovery after photobleaching) showed that removal of raft-targeting signals moderately increased the mobility of HA in the plasma membrane, indicating that the signals influence access of HA to slowly diffusing rafts. However, Myr-Pal–YFP exhibited a much faster mobility compared with HA–Cer, demonstrating that HA and the raft marker do not diffuse together in a stable raft complex for long periods of time.

APA, Harvard, Vancouver, ISO, and other styles

45

CREMAZY, F., E. MANDERS, P. BASTIAENS, G. KRAMER, G. HAGER, E. VANMUNSTER, P. VERSCHURE, T. GADELLAJR, and R. VANDRIEL. "Imaging in situ protein–DNA interactions in the cell nucleus using FRET–FLIM." Experimental Cell Research 309, no. 2 (October 1, 2005): 390–96. http://dx.doi.org/10.1016/j.yexcr.2005.06.007.

Full text

APA, Harvard, Vancouver, ISO, and other styles

46

Long, Yuchen, Yvonne Stahl, Stefanie Weidtkamp-Peters, Marten Postma, Wenkun Zhou, Joachim Goedhart, María-Isabel Sánchez-Pérez, et al. "In vivo FRET–FLIM reveals cell-type-specific protein interactions in Arabidopsis roots." Nature 548, no. 7665 (July 26, 2017): 97–102. http://dx.doi.org/10.1038/nature23317.

Full text

APA, Harvard, Vancouver, ISO, and other styles

47

Venditti, Rossella, Laura Rita Rega, Maria Chiara Masone, Michele Santoro, Elena Polishchuk, Daniela Sarnataro, Simona Paladino, et al. "Molecular determinants of ER–Golgi contacts identified through a new FRET–FLIM system." Journal of Cell Biology 218, no. 3 (January 18, 2019): 1055–65. http://dx.doi.org/10.1083/jcb.201812020.

Full text

Abstract:

ER–TGN contact sites (ERTGoCS) have been visualized by electron microscopy, but their location in the crowded perinuclear area has hampered their analysis via optical microscopy as well as their mechanistic study. To overcome these limits we developed a FRET-based approach and screened several candidates to search for molecular determinants of the ERTGoCS. These included the ER membrane proteins VAPA and VAPB and lipid transfer proteins possessing dual (ER and TGN) targeting motifs that have been hypothesized to contribute to the maintenance of ERTGoCS, such as the ceramide transfer protein CERT and several members of the oxysterol binding proteins. We found that VAP proteins, OSBP1, ORP9, and ORP10 are required, with OSBP1 playing a redundant role with ORP9, which does not involve its lipid transfer activity, and ORP10 being required due to its ability to transfer phosphatidylserine to the TGN. Our results indicate that both structural tethers and a proper lipid composition are needed for ERTGoCS integrity.

APA, Harvard, Vancouver, ISO, and other styles

48

Gratton, Enrico, Elizabeth Hinde, and Michelle A. Digman. "Analysis of FRET Biosensor Distribution in 3D by the Phasor Approach to FLIM." Biophysical Journal 102, no. 3 (January 2012): 234a—235a. http://dx.doi.org/10.1016/j.bpj.2011.11.1289.

Full text

APA, Harvard, Vancouver, ISO, and other styles

49

Borst, Jan Willem, Sergey Laptenok, Ivo van Stokkum, Antonie Visser, and Herbert van Amerongen. "Exploiting the Rise Time of Acceptor Fluorescence by FRET-FLIM in Living Cells." Biophysical Journal 98, no. 3 (January 2010): 580a. http://dx.doi.org/10.1016/j.bpj.2009.12.3150.

Full text

APA, Harvard, Vancouver, ISO, and other styles

50

Padilla-Parra, Sergi, Hervé Lalucque, Marie-Jo Masse, Jean Claude Mével, Nicolas Audugó, Maíté Coppey-Moisan, and Marc Tramier. "In The Quest Of The Best Fluorescent Protein Couple For Quantitative Fret-flim." Biophysical Journal 96, no. 3 (February 2009): 403a. http://dx.doi.org/10.1016/j.bpj.2008.12.2052.

Full text

APA, Harvard, Vancouver, ISO, and other styles

We offer discounts on all premium plans for authors whose works are included in thematic literature selections. Contact us to get a unique promo code!