Hamamatsu-photonics K.k.

Other

Atsuo Miyakawa

Kiyoshi Kamiya

Masahiko Hirano

Shigeru Uchiyama

Norikazu Sugiyama

Eiji Inuzuka

Koji Suzuki

Yoshinori Mizuguchi

Masahumi Oshiro

Hideshi Ohishi

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

Configuration image data representing an external form of an object is stored in advance in a configuration image memory. A two-dimensional image of faint light emitted from the object is detected by a high-sensitivity image pickup means including a two-dimensional photon-counting tube. Faint light image data representing the detected faint light image is accumulated by an adder circuit, and stored in a faint light image memory. The stored faint light image data is superposed on the stored configuration data for each accumulating operation, and based on the superposed image data a superposed image is successivley displayed on a display device.

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Hamamatsu, Japan

Kiyoshi Kamiya