Hamamatsu-photonics K.k.

Riken Corporation

Other

Japan Science and Technology Agency

Atsuo Miyakawa

Masahiko Hirano

Kiyoshi Kamiya

Atsushi Miyawaki

Norikazu Sugiyama

Shigeru Uchiyama

Yoshinori Mizuguchi

Hideshi Oishi

Masafumi Oshiro

Kazuji Matsumoto

Masahiro Hara

Hideji Fujiwake

Iwayo Nakagawa

Hideshi Ohishi

This invention relates to a probe reagent comprising, in order from the N-terminus to the C-terminus, the amino acid sequences of a fluorescent protein I, a peptide capable of terminating protein degradation (i.e., a degradation-terminating peptide), a spacer peptide, a fluorescent protein II, and a protein to be degraded, wherein the protein to be degraded is a protein degraded by the ubiquitin-proteasome system, and the probe reagent is degraded from the C-terminus, but that the degradation of the probe reagent is terminated at the degradation-terminating peptide, a nucleic acid encoding the probe reagent, and use of the probe reagent or the nucleic acid.

Probe reagent for measurement of proteolytic activity

An analyzer for measuring an FRET (fluorescence resonance energy transfer) efficiency of a specimen containing a donor and an acceptor. The analyzer has an illuminator, an optical system, a detector and a calculator. The illuminator emits light for donor excitation and acceptor bleaching. The detector detects fluorescence from the specimen. The calculator calculates the FRET efficiency using the output of the detector. The detector independently detects the fluorescence in wavelength regions. One of the regions has a larger overlap with the fluorescence spectrum of the acceptor than with that of the donor.

Fluorescence resonance energy transfer analyzer

By using an imaging device (), it is possible to determine the concentration of flourescent pigments contained in a target sample (). The imaging device has a plurality of detection wavelength bands. There are prepared a plurality of reference samples, each containing each of the flourescent pigments in a predetermined unit concentration, so as to obtain measurement intensity of flourescent light emitted from each reference sample at each detection wavelength band. The flourescent image of the target sample is imaged at each detection wavelength band by using the imaging device. By using the flourescent intensities obtained from the reference sample and the target sample, calculation is executed so as to obtain the concentration of each of the flourescent pigments in the target sample.

Method and system for measuring the concentrations of fluorescent dyes

A device for measuring an intracellular ion concentration in a living cell into which a fluorescent probe dye has been introduced. A concentration of an ion in the cell is measured based on intensities of fluorescence generated by irradiating the cell with excitation beams, and comparing the intensities of fluorescence emitted by the cell with fluorescence intensities generated from each of 3 reference solutions irradiated with excitation beams. The device is composed of a container means for holding an object to be measured, an excitation beam irradiating means, a fluorescence intensity detecting means, a first processing means for receiving output signals of the fluorescence intensity detecting means and a second processing means for solving simultaneous equations to determine a concentration of the ion to be measured within the cell.

Device for measuring an intracellular ion concentration

In a method for measuring an intracellular ion concentration of the present invention, 4 or more solutions of a fluorescence probe dye and the ions to be measured, 3 or more solutions of an interfering biosubstance and the fluorescent probe dye, and 3 or more solutions of the interfering biosubstance, the fluorescent probe dye and the ions to be measured are prepared. Each solution is irradiated with excitation beams at three different wavelengths, and a fluorescence intensity generated from each solution is measured. Three kinds of equilibrium constants in equilibrium constant equations and 12 kinds of fluorescence coefficients in fluorescence intensity calculating equations are optimized in accordance with a method of successive approximation. With the optimized 15 constants, the equilibrium constant equations and the fluorescence intensity calculation equations are solved to obtain the concentration of ions to be measured. Accordingly, labor for preparing samples is decreased, and the constants are given by a method of successive approximation, which means that the constants are highly reliable, and accordingly the intracellular ion concentration measured based on such highly reliable constants can be accurate.

Method of optimizing equilibrium constants and fluorescence coefficients

A triple view imaging apparatus is provided for measuring quantitative distribution of material or property in a sample. In the triple view imaging apparatus, an optical system receives an original optical image of the sample, separates the original optical image into at least two secondary optical images having different optical properties from one another, and projects the at least two secondary optical images into a single view angle. A single video camera simultaneously picks up the thus projected plurality of secondary optical images as a single composite image and produces image signals representing the light intensities of the plurality of secondary optical images. An image processor receives the image signals and processes the image signals to obtain final image signals representing a relationship between the image signals for respective ones of the plurality of secondary optical images. An image display receives the calculated final image signals and displays a tertiary optical image based on the calculated final image signals which defines quantitative distribution of material or property in the sample.

Triple view imaging apparatus

A fluorescence probe dye is introduced into a cell, and excitation beams at three defferent wavelengths are irradiated to the cell to measure intensities of the fluorescence generated by the excitation beams, corresponding to the three wavelengths. Then, an equilibrium reaction equation for concentrations of the fluorescence probe dye, protein, free ions and their complexes in the cell, and a relationship equation between the fluorescence probe dye, protein, free ions and their complexes are used as simultaneous equations to give concentrations of the respective components. This process can correct interactions among various components of the cell due to bonding among them and a correct ion concentration can be determined.

Method for measurement of intracellular ion concentration using

The present invention relates to an equipment for analyzing the Ca.sup.2+ concentration in the cardiomyocytes, wherein a fluorescent reagent is loaded into the cells to cause intracellular carboxyl groups to be coupled with Ca.sup.2+. The cells are subjected to pulse-excited lights of differing in wavelengths during a vertical blanking period following a predetermined period of time after application of the stimulative signal. The signal causes the cells to generate fluorescent images varying in fluorescence intensity due to the difference in the wavelength of the pulse-excited lights. The resultant images generated are captured by a high-sensitivity television camera and displayed on a video monitor. A plural number of image data are obtained by varying the timing of the stimulating signal with the images stored in image memory. Image data is accessed through a central processing unit which two-dimensionally analyzes the change in the shape of a single cardiomyocyte due to the contraction and the dynamic change in the intracellular Ca.sup.2+ concentration.

Growing community of inventors

Hamamatsu, Japan

Masahiko Hirano