Artículo

Estamos trabajando para incorporar este artículo al repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

The combination of temporal and spectral resolution in fluorescence microscopy based on long-lived luminescent labels offers a dramatic increase in contrast and probe selectivity due to the suppression of scattered light and short-lived autofluorescence. We describe various configurations of a fluorescence microscope integrating spectral and microsecond temporal resolution with conventional digital imaging based on CCD cameras. The high- power, broad spectral distribution and microsecond time resolution provided by microsecond xenon flashlamps offers increased luminosity with recently developed fluorophores with lifetimes in the submicrosecond to microsecond range. On the detection side, a gated microchannel plate intensifier provides the required time resolution and amplification of the signal. Spectral resolution is achieved with a dual grating stigmatic spectrograph and has been applied to the analysis of luminescent markers of cytochemical specimens in situ and of very small volume elements in microchambers. The additional introduction of polarization optics enables the determination of emission polarization; this parameter reflects molecular orientation and rotational mobility and, consequently, the nature of the microenvironment. The dual spectral and temporal resolution modes of acquisition complemented by a posteriori image analysis gated on the spatial, spectral, and temporal dimensions lead to a very flexible and versatile tool. We have used a newly developed lanthanide chelate, Eu-DTPA-cs124, to demonstrate these capabilities. Such compounds are good labels for time-resolved imaging microscopy and for the estimation of molecular proximity in the microscope by fluorescence (luminescence) resonance energy transfer and of molecular rotation via fluorescence depolarization. We describe the spectral distribution, polarization states, and excited-state lifetimes of the lanthanide chelate crystals imaged in the microscope.

Registro:

Documento: Artículo
Título:Temporally and spectrally resolved imaging microscopy of lanthanide chelates
Autor:Vereb, G.; Jares-Erijman, E.; Selvin, P.R.; Jovin, T.M.
Filiación:Department of Molecular Biology, Max Planck Inst. Biophysical Chem., D-37070 Göttingen, Germany
Dept. of Biophysics and Cell Biology, Univ. Medical School of Debrecen, 4012 Debrecen, Hungary
Department of Organic Chemistry, University of Buenos Aires, 1428 Buenos Aires, Argentina
Department of Physics, Univ. Illinois at Urbana-Champaign, Urbana, IL 61801, United States
Department of Molecular Biology, Max Planck Inst. Biophysical Chem., Am Fassberg, D-37077 Göttingen, Germany
Palabras clave:lanthanide; metal chelate; article; crystal structure; fluorescence microscopy; fluorescence polarization; luminescence
Año:1998
Volumen:74
Número:5
Página de inicio:2210
Página de fin:2222
DOI: http://dx.doi.org/10.1016/S0006-3495(98)77930-5
Título revista:Biophysical Journal
Título revista abreviado:Biophys. J.
ISSN:00063495
CODEN:BIOJA
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00063495_v74_n5_p2210_Vereb

Referencias:

  • Altamirano-Bustamante, A., Barnard, G., Kohen, F., Direct time-resolved fluorescence immunoassay for serum oestradiol based on the idiotypic anti-idiotypic approach (1991) J. Immunol. Methods, 138, pp. 95-101
  • Andreev, O.A., Andreeva, A.L., Borejdo, J., Polarization of fluorescently labeled myosin subfragment-1 fully or partially decorating muscle fibers and myofibrils (1993) Biophys. J., 65, pp. 1027-1038
  • Austin, R.H., Chan, S.S., Jovin, T.M., Rotational diffusion of cell surface components by time-resolved phosphorescence anisotropy (1979) Proc. Natl. Acad. Sci. U.S.A., 76, pp. 5650-5654
  • Barnard, G., Kohen, F., Mikola, H., Lovgren, T., The development of non-separation time-resolved fluoroimmunoassays for the measurement of urinary metabolites (1989) J. Biolum. Chemilum., 4, pp. 177-184
  • Beverloo, H.B., Van Schadewijk, A., Bonnet, J., Van Der Geest, R., Runia, R., Verwoerd, N.P., Vrolijk, J., Tanke, H.J., Preparation and microscopic visualization of multicolor luminescent immunophosphors (1992) Cytometry, 13, pp. 561-570
  • Bunzli, J.-C.G., Luminescent probes (1989) Lanthanide Probes in Life, Chemical and Earth Sciences: Theory and Practice, pp. 219-293. , J.-C. G. Bunzli and G. R. Choppin, Eds. Elsevier, New York
  • Burroughs, S.E., Horrocks W.D., Jr., Ren, H., Klee, C.B., Characterization of the lanthanide ion-binding properties of calcineurin-B using laser-induced luminescence spectroscopy (1994) Biochemistry, 33, pp. 10428-10436
  • Carnall, W.T., Fields, P.R., Rajnak, K., Spectral intensities of the trivalent lanthanides and actinides in solution. II. Pm3+, Sm3+, Eu3+, Gd3+, Tb3+, Dy3+, and Ho3+ (1968) J. Chem. Phys., 49, pp. 4412-4423
  • Churchich, J.E., Binding of a fluorescent nucleotide analog to Hsc70: The effect of peptide protein interactions on the luminescence properties of the probe (1995) Eur. J. Biochem., 231, pp. 736-741
  • Ci, Y.X., Yang, X.D., Chang, W.B., Fluorescence labelling with europium chelate of beta-diketones and application in time-resolved fluoroimmunoassays (Tr-Fia) (1995) J. Immunol. Methods, 179, pp. 233-241
  • Clark, I.D., MacManus, J.P., Banville, D., Szabo, A.G., A study of sensitized lanthanide luminescence in an engineered calcium-binding protein (1993) Anal. Biochem., 210, pp. 1-6
  • Clegg, R.M., Fluorescence resonance energy transfer (FRET) (1996) Fluorescence Imaging Spectroscopy and Microscopy, pp. 179-252. , X. F. Wang and B. Herman, Eds. John Wiley & Sons, New York
  • Clegg, R.M., Gadella T.W.J., Jr., Jovin, T.M., Lifetime-resolved fluorescence imaging (1994) Proc. SPIE, 2137, pp. 105-118
  • Dahlen, P., Liukkonen, L., Kwiatkowski, M., Hurskainen, P., Iitia, A., Siitari, H., Ylikoski, J., Lovgren, T., Europium-labeled oligonucleotide hybridization probes: Preparation and properties (1994) Bioconjugate Chem., 5, pp. 268-272
  • Damjanovich, S., Tron, L., Szollosi, J., Zidovetzki, R., Vaz, W.L., Regateiro, F., Arndt-Jovin, D.J., Jovin, T.M., Distribution and mobility of murine histocompatibility H-2Kk antigen in the cytoplasmic membrane (1983) Proc. Natl. Acad. Sci. U.S.A., 80, pp. 5985-5989
  • De Haas, R.R., Verwoerd, N.P., Van Der Corput, M.P., Van Gijlswijk, R.P., Siitari, H., Tanke, H.J., The use of peroxidase-mediated deposition of biotin-tyramide in combination with time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization (1996) J. Histochem. Cytochem., 44, pp. 1091-1099
  • Dexter, D.L., A theory of sensitized luminescence in solids (1953) J. Chem. Phys., 21, pp. 836-850
  • Dickson, E.F., Pollak, A., Diamandis, E.P., Time-resolved detection of lanthanide luminescence for ultrasensitive bioanalytical assays (1995) J. Photochem. Photobiol., 27, pp. 3-19
  • Dix, J.A., Verkman, A.S., Mapping of fluorescence anisotropy in living cells by ratio imaging: Application to cytoplasmic viscosity (1990) Biophys. J., 57, pp. 231-240
  • Drexhage, K.H., Monomolecular layers and light (1970) Sci. Am., 222, pp. 108-119
  • Förster, T., Energiewanderung und Fluoreszenz (1946) Naturwissenschaften, 6, pp. 166-175
  • Gadella T.W.J., Jr., Clegg, R.M., Jovin, T.M., Fluorescence lifetime imaging microscopy: Pixel-by-pixel analysis of phase-modulation data (1994) Bioimaging, 2, pp. 139-159
  • Gadella T.W.J., Jr., Jovin, T.M., Oligomerization of epidermal growth factor receptors on A431 cells studied by time-resolved fluorescence imaging microscopy: A stereochemical model for tyrosine kinase receptor activation (1995) J. Cell Biol., 129, pp. 1543-1558
  • Gadella T.W.J., Jr., Jovin, T.M., Fast algorithms for the analysis of single and double exponential decay curves with a background term: Application to time- Resolved imaging microscopy (1997) Bioimaging, 5, pp. 19-39
  • Gadella T.W.J., Jr., Jovin, T.M., Clegg, R.M., Fluorescence lifetime imaging microscopy (FLIM): Spatial resolution of microstructures on the nanosecond time scale (1993) Biophys. Chem., 48, pp. 221-239
  • Hennink, E.J., De Haas, R., Verwoerd, N.P., Tanke, H.J., Evaluation of a time-resolved fluorescence microscope using a phosphorescent Pt-porphine model system (1996) Cytometry, 24, pp. 312-320
  • Heyduk, E., Heyduk, T., Thiol-reactive luminescent Europium chelates: Luminescence probes for resonance energy transfer distance measurements in biomolecules (1997) Anal. Biochem., 248, pp. 216-227
  • Joshi, N.B., Shamboo, A.E., Distances between functional sites in cardiac sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase: Interlanthanide energy transfer (1988) Eur. J. Biochem., 178, pp. 483-487
  • Jovin, T.M., Arndt-Jovin, D.J., FRET microscopy: Digital imaging of fluorescence resonance energy transfer. Application in cell biology (1989) Cell Structure and Function by Microspectrofluometry, pp. 99-117. , E. Kohen, J. G. Hirschberg, and J. S. Ploem, Eds. Academic Press, London
  • Jovin, T.M., Arndt-Jovin, D.J., Marriott, G., Clegg, R.M., Robert-Nicoud, M., Schormann, T., Distance, wavelength and time: The versatile 3rd dimensions in light emission microscopy (1990) Optical Microscopy for Biology, pp. 575-602. , B. Herman and K. Jacobson, Eds. Wiley-Liss, New York
  • Jovin, T.M., Bartholdi, M., Vaz, W.L., Austin, R.H., Rotational diffusion of biological macromolecules by time-resolved delayed luminescence (phosphorescence, fluorescence) anisotropy (1982) Ann. N.Y. Acad. Sci., 366, pp. 176-196
  • Konig, K., So, P.T., Mantulin, W.W., Tromberg, B.J., Gratton, E., Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress (1996) J. Microsc., 183, pp. 197-204
  • Kwiatkowski, M., Samiotaki, M., Lamminmaki, U., Mukkala, V.M., Landegren, U., Solid-phase synthesis of chelate-labelled oligonucleotides: Application in triple-color ligase-mediated gene analysis (1994) Nucleic Acids Res., 22, pp. 2604-2611
  • Kwok, F., Churchich, J.E., The binding of substrates and inhibitors to the metal center of myoinositol monophosphatase (1994) FEBS Lett., 346, pp. 304-306
  • Lakowicz, J.R., Malak, H., Gryczynski, I., Castellano, F.N., Meyer, G.J., DNA dynamics observed with long lifetime metal-ligand complexes (1995) Biospectroscopy, 1, pp. 163-168
  • Lakowicz, J.R., Szmacinski, H., Nowaczyk, K., Lederer, W.J., Kirby, M.S., Johnson, M.L., Fluorescence lifetime imaging of intracellular calcium in COS cells using Quin-2 (1994) Cell Calcium, 15, pp. 7-27
  • Li, M., Selvin, P.R., Luminescent polyaminocarboxylate chelates of terbium and europium: The effect of chelate structure (1995) J. Am. Chem. Soc., 117, pp. 8132-8138
  • Li, M., Selvin, P.R., Amine-reactive forms of a luminescent diethylenetriaminepentaacetic acid chelate of terbium and europium: Attachment to DNA and energy transfer measurements (1997) Bioconjugate Chem., 8, pp. 127-132
  • Li, L., Szmacinski, H., Lakowicz, J.R., Long-lifetime lipid probe containing a luminescent metal-ligand complex (1997) Biospectroscopy, 3, pp. 155-159
  • Li, L., Szmacinski, H., Lakowicz, J.R., Synthesis and luminescence spectral characterization of long-lifetime lipid metal-ligand probes (1997) Anal. Biochem., 244, pp. 80-85
  • Marriott, G., Clegg, R.M., Arndt-Jovin, D.J., Jovin, T.M., Time resolved imaging microscopy: Phosphorescence and delayed fluorescence imaging (1991) Biophys. J., 60, pp. 1374-1387
  • Marriott, G., Heidecker, M., Diamandis, E.P., Yan-Marriott, Y., Time-resolved delayed luminescence image microscopy using an europium ion chelate complex (1994) Biophys. J., 67, pp. 957-965
  • Martini, J.L., Tetreau, C., Pochon, F., Tourbez, H., Lentz, J.M., Lavalcttc, D., On the mechanism of energy transfer to Tb3+ ions in proteins: A time-resolved luminescence study of the Tb-elastase complex (1993) Eur. J. Biochem., 211, pp. 467-473
  • Mathis, G., Probing molecular interactions with homogeneous techniques based on rare earth cryptates and fluorescence energy transfer (1995) Clin. Chem., 41, pp. 1391-1397
  • Mickols, W., Maestre, M.F., Tinoco I., Jr., Embury, S.H., Visualization of oriented hemoglobin S in individual erythrocytes by differential extinction of polarized light (1985) Proc. Natl. Acad. Sci. U.S.A., 82, pp. 6527-6531
  • Root, D.D., In situ molecular association of dystrophin with actin revealed by sensitized emission immuno-resonance energy transfer (1997) Proc. Natl. Acad. Sci. USA, 94, pp. 5685-5690
  • Selvin, P.R., Fluorescence resonance energy transfer (1995) Methods Enzymol., 246, pp. 300-334
  • Selvin, P.R., Lanthanide-based resonance energy transfer (1996) IEEE JSTQE: Lasers Biol., 2, pp. 1077-1087
  • Selvin, P.R., Hearst, J.E., Luminescence energy transfer using a terbium chelate: Improvements on fluorescence energy transfer (1994) Proc. Natl. Acad. Sci. U.S.A., 91, pp. 10024-10028
  • Selvin, P.R., Jancarik, J., Li, M., Hung, L.W., Crystal structure and spectroscopic characterization of a luminescent europium chelate (1996) Inorg. Chem., 35, pp. 700-705
  • Selvin, P.R., Rana, T.M., Hearst, J.E., Luminescence resonance energy transfer (1994) J. Am. Chem. Soc., 116, pp. 6029-6030
  • Seveus, L., Vaisala, M., Syrjanen, S., Sandberg, M., Kuusisto, A., Harju, R., Salo, J., Soini, E., Time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization (1992) Cytometry, 13, pp. 329-338
  • Szmacinski, H., Terpetschnig, E., Lakowicz, J.R., Synthesis and evaluation of Ru-complexes as anisotropy probes for protein hydrodynamics and immunoassays of high-molecular-weight antigens (1996) Biophys. Chem., 62, pp. 109-120
  • Szöllösi, J., Tron, L., Damjanovich, S., Helliwell, S.H., Arndt-Jovin, D., Jovin, T.M., Fluorescence energy transfer measurements on cell surfaces: A critical comparison of steady-state fluorimetric and flow cytometric methods (1984) Cytometry, 5, pp. 210-216
  • Terpetschnig, E., Szmacinski, H., Lakowicz, J.R., Fluorescence polarization immunoassay of a high-molecular-weight antigen based on a long-lifetime Ru-ligand complex (1995) Anal. Biochem., 227, pp. 140-147
  • Terpetschnig, E., Szmacinski, H., Lakowicz, J.R., Fluorescence polarization immunoassay of a high-molecular-weight antigen using a long wavelength-absorbing and laser diode-excitable metal-ligand complex (1996) Anal. Biochem., 240, pp. 54-59
  • Terpetschnig, E., Szmacinski, H., Malak, H., Lakowicz, J.R., Metal-ligand complexes as a new class of long-lived fluorophores for protein hydrodynamics (1995) Biophys. J., 68, pp. 342-350
  • Verkman, A.S., Armijo, M., Fushimi, K., Construction and evaluation of a frequency-domain epifluorescence microscope for life- Time and anisotropy decay measurements in subcellular domains (1991) Biophys. Chem., 40, pp. 117-125
  • Verwoerd, N.P., Hennink, E.J., Bonnet, J., Van Der Geest, C.R., Tanke, H.J., Use of ferro-electric liquid crystal shutters for time-resolved fluorescence microscopy (1994) Cytometry, 16, pp. 113-117
  • Walters, J.D., Johnson, J.D., Terbium as a luminescent probe of metal-binding sites in protein kinase C (1990) J. Biol. Chem., 265, pp. 4223-4226
  • Youn, H.J., Terpetschnig, E., Szmacinski, H., Lakowicz, J.R., Fluorescence energy transfer immunoassay based on a long-lifetime luminescent metal-ligand complex (1995) Anal. Biochem., 232, pp. 24-30
  • Zidovetzki, R., Johnson, D.A., Arndt-Jovin, D.J., Jovin, T.M., Rotational mobility of high-affinity epidermal growth factor receptors on the surface of living A431 cells (1991) Biochemistry, 30, pp. 6162-6166

Citas:

---------- APA ----------
Vereb, G., Jares-Erijman, E., Selvin, P.R. & Jovin, T.M. (1998) . Temporally and spectrally resolved imaging microscopy of lanthanide chelates. Biophysical Journal, 74(5), 2210-2222.
http://dx.doi.org/10.1016/S0006-3495(98)77930-5
---------- CHICAGO ----------
Vereb, G., Jares-Erijman, E., Selvin, P.R., Jovin, T.M. "Temporally and spectrally resolved imaging microscopy of lanthanide chelates" . Biophysical Journal 74, no. 5 (1998) : 2210-2222.
http://dx.doi.org/10.1016/S0006-3495(98)77930-5
---------- MLA ----------
Vereb, G., Jares-Erijman, E., Selvin, P.R., Jovin, T.M. "Temporally and spectrally resolved imaging microscopy of lanthanide chelates" . Biophysical Journal, vol. 74, no. 5, 1998, pp. 2210-2222.
http://dx.doi.org/10.1016/S0006-3495(98)77930-5
---------- VANCOUVER ----------
Vereb, G., Jares-Erijman, E., Selvin, P.R., Jovin, T.M. Temporally and spectrally resolved imaging microscopy of lanthanide chelates. Biophys. J. 1998;74(5):2210-2222.
http://dx.doi.org/10.1016/S0006-3495(98)77930-5