JPH04125450A - Fluorescence spectrophotometer - Google Patents

Abstract

PURPOSE:To enable simultaneous measurement of function mechanism estimation of blood platelet chemical and failure detection of blood platelet function by a constitution for leading fluorescent light from a specimen and specimen transmitted light to a fluorescence spectrometer by means of a combination of plane mirrors on a side of transmission of specimen application light. CONSTITUTION:Monochromatic light emitted from an excitation spectrometer 2 radiates a specimen vessel 3, and fluorescent light generated from the specimen passes through a beam splitter 12 and is subjected to wavelength selection in a fluorescence spectrometer 4. Then its intensity is measured by a sensor 5. Light which has transmitted through the specimen is reflected on a mirror A10 and a mirror B11, further reflected on the beam splitter 12 and is subjected to wavelength selection in the spectrometer 4, and then its intensity is measured by the sensor 5. Thus a change in fluorescence intensity from the specimen and change in intensity of the specimen transmitted light can be simultaneously measured by a single apparatus, so that simultaneous measurement of estimation of function mechanism of blood platelet chemical and failure detection of blood platelet function is possible.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention is directed to the treatment of intracellular Ca, which is carried out in medicine, pharmacy, etc.
Regarding measurement, this invention relates to a spectrofluorometer that can simultaneously measure Caz+ concentration changes and platelet aggregation measurements.

[Conventional technology]

In order to elucidate the cellular information transmission mechanism, evaluate the mechanism of action of antiplatelet agents, and detect platelet function abnormalities, changes in intracellular Ca' concentration and measurement of platelet aggregation are essential.
A spectrofluorometer is used as a method to measure changes in a2+ concentration. Spectrophotometer is also used as a method to measure changes in platelet aggregation.

A platelet aggregation measuring device is used.

[Problem to be solved by the invention]

However, Caz+ concentration change measurement measures the intensity change of fluorescence emitted from the sample, and platelet aggregation change measurement measures the intensity change of the light transmitted through the sample, so with a spectrofluorometer, the light transmitted through the sample is , cannot be measured. Similarly, spectrophotometers and platelet aggregation measuring devices can measure changes in the intensity of light that passes through a sample, but cannot measure changes in the intensity of fluorescence. By this, Ca2
+There was a problem in that it was not possible to simultaneously measure changes in concentration and changes in platelet aggregation ability. Furthermore, since the light intensity of transmitted light is orders of magnitude stronger than fluorescence intensity, there is a problem in that simultaneous measurement and weak changes in transmitted light cannot be measured unless the transmitted light is attenuated.

The present invention enables a spectrofluorometer to measure fluorescence intensity changes and transmittance changes of a sample at the same time in the same intensity state by combining a flat mirror after the light irradiated onto the sample passes through the sample. This enables simultaneous measurement of the mechanism of action of antiplatelet agents and the detection of platelet function abnormalities due to changes in platelet aggregation.

[Means to solve the problem]

To achieve the above purpose, there is a light source, an excitation side spectrometer and excitation wavelength drive system that separates the light from this light source and irradiates the sample with monochromatic light, a fluorescence side spectrometer and fluorescence wavelength that separates the fluorescence emitted from the sample. Drive system, detector for detecting the emitted light from this fluorescence side spectrometer, amplification of the signal output from the detector, A/
In a spectrofluorometer that consists of a signal amplifier that performs D conversion and a computer that processes the signal, a flat mirror is used on the side where the light that irradiates the sample passes through the sample to combine the fluorescence emitted from the sample and the light that passes through the sample. The transmitted light is guided to a fluorescence spectrometer.

[Effect]

The monochromatic light separated by the excitation side spectrometer is irradiated onto the sample,
The fluorescence emitted from the sample is introduced into the fluorescence side spectrometer,
The light that has passed through the sample is introduced into a fluorescence-side spectrometer via a mirror system and is spectrally analyzed in the same manner as the fluorescence described above. The light is detected by a detector and output as a signal. The output signal is data-processed by a computer. At this time,
The monochromatic light that irradiates the fluorescence and transmitted light sample and the monochromatic light that is separated by the fluorescence side spectrometer are different, so it is necessary to control the excitation wavelength drive system and fluorescence wavelength drive system and set the wavelength using a computer. .

By data processing the signal from the detector in synchronization with this setting, it is possible to distinguish whether the light introduced into the detector is fluorescence or transmitted light. Furthermore, by combining plane mirrors, the transmitted light intensity can be attenuated and measured at the same level as the fluorescence intensity. This makes it possible to simultaneously measure changes in fluorescence and transmitted light of the sample.

〔Example〕

An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 shows the configuration of a typical spectrophotometer. The light beam emitted from the light source 1 is converted into monochromatic light by the excitation side spectrometer 2, and is irradiated onto the sample container 3. After the wavelength of the fluorescence emitted from the sample is selected by the fluorescence side spectrometer 4, it is sent to the detector 5.
Its strength is measured by The signal output from the detector 5 is amplified and A/D converted by a signal amplifier 8, and then sent to a computer 9. Further, the wavelength of the excitation side spectrometer 2 is controlled by the computer 9 via the excitation wavelength drive system 6, and similarly, the wavelength of the fluorescence side spectrometer 4 is controlled by the computer 9 via the fluorescence wavelength drive system 7. ing.

FIG. 2 shows the basic principle of the invention.

The monochromatic light emitted from the excitation side spectrometer 2 is irradiated onto the sample container 3 . The fluorescence emitted from the sample passes through the beam splitter 12 and has its wavelength selected by the fluorescence spectrometer 4, and then its intensity is measured by the detector 5. The transmitted light that has passed through the sample is reflected by the mirror A10 and the mirror Bll, and is further reflected by the beam splitter 12, and after being wavelength-selected by the fluorescence side spectrometer 4, its intensity is measured by the detector 5.

〔Effect of the invention〕

According to the present invention, it is possible to simultaneously measure changes in the intensity of fluorescence emitted from a sample and changes in the intensity of light passing through the sample with one device. It will also lead to the elucidation of the intracellular transmission mechanism, which is effective in simultaneously detecting platelet function abnormalities due to changes in platelet aggregation ability.

[Brief explanation of drawings]

FIG. 1 is a diagram showing a device configuration necessary to realize an embodiment of the present invention, and FIG. 2 is a diagram showing the basic principle of the present invention. DESCRIPTION OF SYMBOLS 1... Light source, 2... Excitation side spectrometer, 3... Sample container, 4... Fluorescence side spectrometer, 5... Detector, 6... Excitation wavelength drive system, 7... Fluorescence wavelength drive system, 8... Signal amplifier, 9... Computer, 10... Plane mirror A, 11... Plane mirror B, 12... Beam splitter.

Claims (1)

[Claims] 1. A light source, an excitation side spectrometer and excitation wavelength drive system that separates the light from this light source and irradiates the sample with monochromatic light, a fluorescence side spectrometer and fluorescence wavelength drive system that separates the fluorescence emitted from the sample, and this fluorescence A spectrofluorometer consists of a detector that detects the emitted light from the side spectrometer, a signal amplifier that amplifies and A/D converts the signal output from the detector, and a computer that processes the signal. A mirror system is provided on the transmitting side of the light to enable simultaneous measurement of time changes in fluorescence intensity and time changes in absorption of the sample, and the mirror system is a combination of plane mirrors to attenuate the intensity of the transmitted light. Spectrofluorometer.