Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Földes-Papp, Z; Demel, U; Tilz, GP.
Detection of single molecules: solution-phase single-molecule fluorescence correlation spectroscopy as an ultrasensitive, rapid and reliable system for immunological investigation.
J Immunol Methods. 2002; 260(1-2):117-124
Doi: 10.1016/S0022-1759(01)00537-3
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- Führende Autor*innen der Med Uni Graz
- Földes-Papp Zeno
- Tilz Gernot
- Co-Autor*innen der Med Uni Graz
- Demel Ulrike
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- Abstract:
- More sensitive techniques in molecular and clinical immunology are essential for the development of reproducible profiles. We have developed a novel methodology named solution-phase single-molecule fluorescence correlation spectroscopy (SPSM-FCS) that fulfils these demands. It is based on the quantification of the probability density of single molecule events in solution is necessary. For example, the Brownian motion of the fluorophore rhodamine-green is detected. Counting about 100,000 photon counts per second and per molecule permits the identification of one single compound. In order to study the applicability of SPSM-FCS in immunology, we have detected and identified a larger nonfluorescent substance in a very complex mixture. The 'unknown' molecules studied were the autoantibodies in serum samples directed against the antigen alpha 3 chain of type IV collagen. In both systems, we were able to characterize the probability density of single fluorescent molecules by means of the averaged absolute molecule number without any calibration. The specific molecules exhibited a Poisson distribution in solution in terms of their 'critical' bulk concentration below about 1 nM. This proof of principle indicates how the SPSM-FCS methodology could be used in immunoassays.
- Find related publications in this database (using NLM MeSH Indexing)
- Animals -
- Antibodies - analysis
- Antibody Formation - analysis
- Humans - analysis
- Immunoassay - methods
- Sensitivity and Specificity - methods
- Spectrometry, Fluorescence - methods
- Find related publications in this database (Keywords)
- single-molecules
- ultrasensitive detection in molecular and clinical immunology
- probability density of single molecules in
- solution-phase
- quantification of Brownian motions
- single (solution)-phase single-molecule fluorescence correlation spectroscopy