Covalent immobilization of oligonucleotides to microparticles. Quantitation of hybridization by time-resolved fluorescence detection on a single particle

1996 ◽  
Vol 61 (s1) ◽  
pp. 107-109
Author(s):  
Harri Hakala ◽  
Pia Heinonen ◽  
Antti Iitia ◽  
Harri Lönnberg
Keyword(s):  
Fluorescence Detection ◽  
Single Particle ◽  
Covalent Immobilization ◽  
Time Resolved Fluorescence ◽  
Time Resolved

scholarly journals Development, Characterization, and Application of a Versatile Single Particle Detection Apparatus for Time-Integrated and Time-Resolved Fluorescence Measurements—Part II: Experimental Evaluation

2009 ◽  
Vol 2009 ◽  
pp. 1-14
Author(s):  
Xihong Wu ◽  
J. A. Merten ◽  
N. Omenetto ◽  
B. W. Smith ◽  
J. D. Winefordner
Keyword(s):  
Single Particle ◽  
Narrow Beam ◽  
Time Resolved Fluorescence ◽  
Particle Detection ◽  
Time Resolved ◽  
Endogenous Fluorophores ◽  
Fluorescence Measurements ◽  
On Line ◽  
Single Particle Detection ◽  
Speed Mode

This paper describes the experimental realization and characterization of a versatile single particle detection apparatus. The system utilizes a novel particle beam inlet that can serve as either an on-line particle concentrator (i.e., all diameters confined in a narrow beam) or as a segregator (i.e., selected diameters confined in a narrow beam) and can be operated in a high-speed mode as well as in a low-speed mode, thus allowing different interaction times between the particles and the laser beam. An aerodynamic sizing technique has been incorporated into the system to provide rapid, real-time, and high-resolution sizing. Parameters such as transmission efficiency and size-segregation efficiency have been measured. The performance of the instrument has been demonstrated by on-line detection of spectrally resolved and time resolved fluorescence detection from airborne dye-doped particles and aerosolized endogenous fluorophores found in biological agents.

Sensitive miniature single-particle immunoassay of prostate-specific antigen using time-resolved fluorescence

2003 ◽  
Vol 482 (2) ◽  
pp. 157-164 ◽  
Author(s):  
Harri Härmä ◽  
Anne-Maria Pelkkikangas ◽  
Tero Soukka ◽  
Petri Huhtinen ◽  
Saila Huopalahti ◽  
...  
Keyword(s):  
Prostate Specific Antigen ◽  
Single Particle ◽  
Specific Antigen ◽  
Time Resolved Fluorescence ◽  
Time Resolved

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.

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