Time-Resolved Fluorescence Detection of Oligonucleotide Hybridization on a Single Microparticle:  Covalent Immobilization of Oligonucleotides and Quantitation of a Model System

1997 ◽  
Vol 8 (2) ◽  
pp. 232-237 ◽  
Author(s):  
Harri Hakala ◽  
Harri Lönnberg
Keyword(s):  
Fluorescence Detection ◽  
Covalent Immobilization ◽  
Model System ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Oligonucleotide Hybridization

Characterization of Plasma-Modified Fluoropolymer Surfaces Using Steady-State and Time-Resolved Fluorescence Spectroscopy

1994 ◽  
Vol 48 (5) ◽  
pp. 630-637 ◽  
Author(s):  
Ming Li ◽  
Michaeleen L. Pacholski ◽  
Frank V. Bright
Keyword(s):  
Steady State ◽  
Fluorescence Spectroscopy ◽  
Sulfonyl Chloride ◽  
Sodium Dodecylsulfate ◽  
Covalent Immobilization ◽  
Ambient Conditions ◽  
Time Resolved Fluorescence ◽  
Decay Kinetics ◽  
Time Resolved

Poly(hexafluoropropylene-co-tetrafluoroethylene) (FEP) has been widely used in biotechnology because of its unique surface properties and biocompatibility. Recent work from our group has shown that plasma discharge-modified FEP can be used as the substratum for development of a very stable immunosensor. This result has prompted us to study further this new surface under ambient conditions. In this paper, we report on the covalent immobilization of a pyrene residue (-Py) onto FEP-APS (FEP-aminopropyl silane) surfaces and the characterization of FEP-APS-Py using steady-state and time-resolved fluorescence spectroscopy. Among the immobilization schemes tested, we found that the covalent coupling of pyrene-sulfonyl chloride to FEP-APS is the easiest and yields the most photostable FEP-APS-Py derivative. Steady-state emission spectra of FEP-APS-Py in contact with H2O, β-cyclodextrin (β-CD), and sodium dodecylsulfate (SDS) aqueous solutions differ considerably from those of Py-SO3 in solution. Time-resolved fluorescence spectroscopy of FEP-APS-Py demonstrates that the decay kinetics are strongly affected by the presence of ionic quenchers and molecular oxygen, as well as β-CD and SDS. The results are consistent with the suggestion that the APS-Py moiety undergoes a slow time-dependent reconfiguration at the FEP/APS interface.

Detection of Escherichia coli rRNA using target amplification and time-resolved fluorescence detection

1991 ◽  
Vol 5 (6) ◽  
pp. 467-472 ◽  
Author(s):  
Charlene E. Bush ◽  
Kurt M. Vanden Brink ◽  
David G. Sherman ◽  
W. Richard Peterson ◽  
Laura A. Beninsig ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescence Detection ◽  
Time Resolved Fluorescence ◽  
Time Resolved

An automated PCR platform with homogeneous time-resolved fluorescence detection and dry chemistry assay kits

2008 ◽  
Vol 374 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Virve Hagren ◽  
Piia von Lode ◽  
Anniina Syrjälä ◽  
Tero Soukka ◽  
Timo Lövgren ◽  
...  
Keyword(s):  
Fluorescence Detection ◽  
Time Resolved Fluorescence ◽  
Time Resolved

Time-resolved fluorescence detection of enzyme-amplified lanthanide luminescence for nucleic acid hybridization assays

1991 ◽  
Vol 37 (9) ◽  
pp. 1506-1512 ◽  
Author(s):  
E F Templeton ◽  
H E Wong ◽  
R A Evangelista ◽  
T Granger ◽  
A Pollak
Keyword(s):  
Alkaline Phosphatase ◽  
Nucleic Acid ◽  
Fluorescence Detection ◽  
Dna Hybridization ◽  
Nucleic Acid Hybridization ◽  
Time Resolved Fluorescence ◽  
Dot Blot ◽  
Time Resolved ◽  
Lanthanide Luminescence ◽  
Hybridization Assays

Abstract A new nonisotopic detection method based on time-resolved fluorescence for nucleic acid hybridization assays with alkaline phosphatase labels has been developed: enzyme-amplified lanthanide luminescence (EALL). EALL combines the amplification of an enzyme label with the sensitivity and background elimination of time-resolved fluorescence detection of lanthanide ion luminescence. The detection system for alkaline phosphatase makes use of a phosphorylated salicylic acid derivative that, upon dephosphorylation, gives a product capable of forming a luminescent terbium chelate. We demonstrate DNA hybridization assays by using two substrates, one for membrane and one for solution-based formats. Using the substrate that produces a more adhesive product allows performance of dot-blot and Southern blot assays on nylon membranes; results can be recorded with a time-resolved photographic camera system, or with an ultraviolet transilluminator-based system. Less than 4 pg of target sequence can be detected in a dot-blot assay after incubation with substrate for 2-4 h. DNA microwell-plate hybridization assays with the more soluble substrate/product pair can be quantified with time-resolved fluorescence plate readers, giving a similar detection sensitivity. EALL is thus a practical time-resolved fluorescence-based alternative to other detection systems for DNA hybridization assays.

scholarly journals Gene expression analysis with an integrated CMOS microarray by time-resolved fluorescence detection

2011 ◽  
Vol 26 (5) ◽  
pp. 2660-2665 ◽  
Author(s):  
Ta-chien D. Huang ◽  
Sunirmal Paul ◽  
Ping Gong ◽  
Rastislav Levicky ◽  
John Kymissis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Fluorescence Detection ◽  
Gene Expression Analysis ◽  
Time Resolved Fluorescence ◽  
Time Resolved
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