Fluorescence-intensity distribution analysis and its application in biomolecular detection technology.
Proc Natl Acad Sci U S A
; 96(24): 13756-61, 1999 Nov 23.
Article
in En
| MEDLINE
| ID: mdl-10570145
ABSTRACT
A methodology, fluorescence-intensity distribution analysis, has been developed for confocal microscopy studies in which the fluorescence intensity of a sample with a heterogeneous brightness profile is monitored. An adjustable formula, modeling the spatial brightness distribution, and the technique of generating functions for calculation of theoretical photon count number distributions serve as the two cornerstones of the methodology. The method permits the simultaneous determination of concentrations and specific brightness values of a number of individual fluorescent species in solution. Accordingly, we present an extremely sensitive tool to monitor the interaction of fluorescently labeled molecules or other microparticles with their respective biological counterparts that should find a wide application in life sciences, medicine, and drug discovery. Its potential is demonstrated by studying the hybridization of 5'-(6-carboxytetramethylrhodamine)-labeled and nonlabeled complementary oligonucleotides and the subsequent cleavage of the DNA hybrids by restriction enzymes.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA
/
Microscopy, Confocal
/
Microscopy, Fluorescence
Type of study:
Diagnostic_studies
Language:
En
Journal:
Proc Natl Acad Sci U S A
Year:
1999
Document type:
Article
Affiliation country: