MC-1 A new generation of time-resolved fluoroimmunoassays with europium chelates as labels

1988 ◽  
Vol 21 (6) ◽  
pp. 381
Author(s):  
E.P. Diamandis ◽  
R.A. Evangelista ◽  
A. Pollak ◽  
E.F. Templeton ◽  
J.A. Lowden
2004 ◽  
Vol 850 ◽  
Author(s):  
S. Benson ◽  
D. R. Douglas ◽  
H. F. Dylla ◽  
J. Gubeli ◽  
K. Jordan ◽  
...  

ABSTRACTWe describe a > 100 Watt broadband THz source that takes advantage of the relativistic enhancement of the radiation from accelerating electrons according to the formula assigned the name of Sir Joseph Larmor[1, 2]. This is in contrast to the typical 1 milliwatt sources available in a laboratory. Specifically, for relativistic electrons the emission is enhanced by the fourth power of the increase in mass. Thus for 100 MeV electrons, for which the mass increases by a factor of ∼ 200, the enhancement is > 109. The experiments use a new generation of light source called an energy recovery linac (ERL) [3], in which bunches of electrons circulate once, but in which their energy is recovered. In such a machine the electron bunches can be very much shorter than those, say, in storage rings or synchrotrons.The Jefferson Lab facility operates in new limits of emission from relativistic particles involving both multiparticle coherence and near-field emission in which the velocity (Coulomb) term in the classical electrodynamical theory becomes as important as the acceleration term (synchrotron radiation).The sub-picosecond pulses of light offer unique capabilities in 2 specific areas, namely time-resolved dynamics, and imaging. High resolution THz spectroscopy has recently revealed sharp vibrational modes for many materials including malignant tissue, proteins, DNA, pharmaceuticals and explosive materials. Energetically the THz range embraces superconducting bandgaps, and regions of intense interest in the understanding of systems in which correlated motions of electrons are important, such as colossal magneto-resistive and high-Tc materials. The very high power levels of the new source will allow non-linear effects to be observed as well as the creation of novel states of materials, including electric-field driven localization[4]. We will give examples of existing work in these areas and present opportunities afforded by the new source.


1997 ◽  
Vol 43 (10) ◽  
pp. 1937-1943 ◽  
Author(s):  
Timo Lövgren ◽  
Pia Heinonen ◽  
Päivi Lehtinen ◽  
Harri Hakala ◽  
Johanna Heinola ◽  
...  

Abstract Future immunoassays and nucleic acid hybridization assays will be performed in miniaturized formats that utilize microchips or microparticles. This will require a sensitive detection technology that allows spatial resolution. By using fluorescent europium chelates and time-resolved microfluorometry, one can detect 11 000 europium molecules on individual microparticles. In a miniaturized noncompetitive immunoassay of prostate-specific antigen (PSA), we quantitatively detected 5 ng/L (0.05 amol per particle) of the analyte on an individual microparticle with excellent precision over the whole measurement range (CV <10%). Using a hybridization assay, we also could detect the ΔF508 mutation for cystic fibrosis on individual microparticles. Consequently, fluorescent lanthanide chelate labels and time-resolved microfluorometry qualify as the next generation of technology in this field.


2011 ◽  
Vol 400 (8) ◽  
pp. 2487-2497 ◽  
Author(s):  
Christoph Wagner ◽  
Wolfgang Buchegger ◽  
Michael Vellekoop ◽  
Martin Kraft ◽  
Bernhard Lendl

1996 ◽  
Vol 44 (10) ◽  
pp. 1091-1099 ◽  
Author(s):  
R R de Haas ◽  
N P Verwoerd ◽  
M P van der Corput ◽  
R P van Gijlswijk ◽  
H Siitari ◽  
...  

The application of europium chelates as delayed fluorescent labels in FISH and immunocytochemistry is hampered by their relatively low quantum yield. To increase the intensity of the delayed fluorescence, we have used a recently introduced peroxidase-mediated amplification system. This system results in a large accumulation of biotin-tyramide, which is detected using streptavidin-europium chelate as label. Optimal staining conditions were evaluated for the immunocytochemical detection of vimentin in cryosections of rat liver, for DNA in situ hybridization (alphoid type probes and 40-KB cosmid probes), and for RNA in situ hybridization (detection of 28S ribosomal RNA, human elongation factor mRNA, and luciferase mRNA). Using a time-resolved fluorescence microscope, intense europium fluorescence was obtained in all these applications when the tyramide amplification system was applied. The signals were strong enough to be observed by eye using the microscope in the time-delayed mode. The routine application of this technique for localization and quantization of antigens or nucleic acid sequences in tissue exhibiting strong autofluorescence is discussed.


2015 ◽  
Author(s):  
Daniel A Keedy ◽  
Lillian R Kenner ◽  
Matthew Warkentin ◽  
Rahel Woldeyes ◽  
Michael C Thompson ◽  
...  

Determining the interconverting conformations of dynamic proteins in atomic detail is a major challenge for structural biology. Conformational heterogeneity in the active site of the dynamic enzyme cyclophilin A (CypA) has been previously linked to its catalytic function. Here we compare the conformational ensembles of CypA by fixed-target X-ray free electron laser (XFEL) crystallography and multitemperature synchrotron crystallography. The “diffraction-before-destruction” nature of XFEL experiments provides a radiation-damage-free view of the functionally important alternative conformations of CypA. We monitored the temperature dependences of these alternative conformations with eight synchrotron datasets spanning 100-310 K. Multiconformer models show that many alternative conformations in CypA are populated above, but not below, the glass transition temperature (~200 K) and reveal abrupt changes in protein flexibility that provide all-atom insight into conformational coupling. Together, our XFEL data and multitemperature analyses motivate a new generation of time-resolved experiments to structurally characterize the dynamic underpinnings of protein function.  


1983 ◽  
Vol 29 (1) ◽  
pp. 65-68 ◽  
Author(s):  
E Soini ◽  
H Kojola

Abstract Pulsed-light time-resolved fluorometry of lanthanide chelates has proved to be very sensitive for use with nonisotopic immunoassays. We describe a manually operated fluorometer with a conventional xenon flashtube. Sensitivity for 1-s determinations is similar to that of radioisotopic methods.


1965 ◽  
Vol 43 (5) ◽  
pp. 1680-1687 ◽  
Author(s):  
M. L. Bhaumik ◽  
L. J. Nugent

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