Other

Pekka H&auml;nninen

Erkki Soini

Juhani Soini

A method for measuring the end point and for monitoring the real time kinetics of a bioaffinity reaction in biological fluids and suspensions, employing microparticles as bioaffinity binding solid phase, biospecific reagent labelled with a fluorescent label and a fluorescence detection system which is based on two-photon fluorescence excitation, contacting the analyte, the labelled reagent and the solid phase simultaneously, focusing a two-photon exciting laser beam into the reaction suspension and measuring the fluorescence signal emitted by the microparticles from one particle at a time when they randomly float through the focal volume of the laser beam. In this method the signal is monitored kinetically to obtain information about the analyte concentration before the reaction approaches the highest point of the response. Since the growth rate of the signal intensity is directly proportional to the analyte concentration, the analyte concentration can be predicted in the initial phase of the reaction. The growth rate monitoring also predicts whether the analyte concentration is higher than the binding capacity of the reagent and whether the reaction will be continued over the highest point of the response curve.

Biospecific, two photon excitation, fluorescence detection and device

A homogeneous biospecific assay method for an analyte in solution or in a biological suspension, in which a biospecific reagent competitively binding an analyte and a ligand labeled with a fluorescent molecule, is reacted with and bound to a solid phase, and in which the free labeled ligand is extracted is excited with two-photon excitation by focusing a laser beam suitable for two-photon excitation into the sample volume; and the concentration of the analyte is calculated based on the photon emission contributed by the free labeled ligand.

Homogeneous biospecific assay using a solid phase, two-photon excitation and confocal fluorescence detection

A method for quantitatively measuring nucleic acid amplification reactions, especially the polymerase chain reaction, employing microparticles as hybridization solid phase, a probe sequence labeled with a fluorescent label and a fluorescence detection system which is based on two-photon fluorescence excitation, contacting all the amplification reaction components and the solid phase simultaneously in a closed cuvette, performing the amplification reactions in the same cuvette, focusing a two-photon exciting laser beam into the cuvette during the amplification cycles and measuring the fluorescence signal emitted by the microparticles from one particle at a time when they randomly float through the focal volume of the laser beam. The features of this invention allow a method and device for performing a fast quantitative nucleic acid amplification assay of single or multiple target sequences in a very small closed sample volume.

Method and a device for monitoring nucleic acid amplification reactions

This invention is thus related to a biospecific multiparameter assay employing single molecule detection. In particular, this invention is related to a new method for determination of several different biomolecules simultaneously in the same reaction solution. The method uses a fluorescent label for labelling biospecific primary probes and a combination of other labels, bound to secondary probes. Complexes are formed comprising primary probes, analyte molecules and secondary probes in the biospecific reaction. These single molecule complexes are counted selectively while discriminating the signals from other fluorescent molecules using confocal fluorometry or two-photon excitation fluorometry including an auto- and cross-correlator for the signals obtained from said fluorescent labels.

Growing community of inventors

Turku, Finland

Pekka Hänninen

Growing community of inventors

Turku, Finland

Pekka H&auml;nninen

Pekka Hänninen