JPH07128239A - Optical exciting device for caged compound - Google Patents

Abstract

PURPOSE:To observe a reaction in more detail by finely controlling emitting light quantity when a light is emitted to a caged compound. CONSTITUTION:A sample 10 is put on a sample base, and an observing light is emitted from the lower part of the sample base by an observing light source 20, and converged to the sample 10 by a condenser lens 22. The observing light transmitted by the sample 10 is collected by an objective lens 24 and fluoroselectively detected by a detector 26. In an optical exciting device for exciting a caged compound contained in the sample 10 and releasing a reactant, a light source 2 whose emitting power is controlled by a light source controller 4 is provided, and the light from the light source 2 is guided to a wavelength selecting device 6 by a convergence lens 12. A shutter 14 and a slit 16 are arranged between the convergence lens 12 and the wavelength selecting device 6, and the emitting timing and emitting time are controlled by the shutter 14. The emitting light whose wavelength is selected by the wavelength selecting device 6 is emitted to the sample 10 by a convergence lens 18.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a cadide compound in the fields of analytical chemistry, physical chemistry, biophysics, biochemistry, material science, etc. The present invention relates to a device for photoexcitation.

[0002]

2. Description of the Related Art One reaction substance in a chemical reaction system, a reaction substance for measuring a biological activity, or a substance related to chemiluminescence or bioluminescence starts a reaction when they are injected into a reaction system or an object. To do. For example, when a reaction product is injected into a chemical reaction system to start a reaction, it takes time for the reaction product to uniformly diffuse into the chemical reaction system, and the reaction system as a whole cannot start the reaction uniformly. Therefore, it has been studied to inject the reaction product as a cadide compound into the reaction system and to release the reaction product from the cadide compound by light irradiation to start the reaction.

[0003] In addition, since calcium ions act as regulators of various functions in the living body, it has been performed to observe the reaction by adding calcium ions into cells. In this case, when calcium ions are added to the cells, the reaction starts from that point, and only the reaction to the added amount of calcium is observed. Also in this case, calcium ions are added to the cell as a cadide compound,
It is known that calcium ions are released from a cadide compound by irradiation with light to start the reaction.

[0004]

A method of dissociating and releasing a reaction product by light irradiation using a cadide compound,
Although the measurement is carried out after the concentration of the reactant released from the cadide compound after the light irradiation is made constant, sufficient information cannot be obtained only by observing the reaction of the system after making the concentration constant. It is necessary to control the concentration of the reactants in real time.

Therefore, it is an object of the present invention to make it possible to finely control the amount of irradiation light when irradiating the cadide compound with light so that more detailed reaction can be observed. The present invention also allows multiple reactions with cadide compounds to occur in one reaction system.

[0006]

SUMMARY OF THE INVENTION The present invention is a device for irradiating a caged compound installed in an optical measuring device with excitation light, wherein a light source part, an irradiation power, an irradiation timing and an irradiation time of the light source part. A light source controller for controlling and a control interface for exchanging signals between the light source controller and an external device are provided. In a preferred embodiment, the light source unit further includes a wavelength selection device that selects the wavelength of the light to be emitted to the caded compound, and the control interface also exchanges signals between the wavelength selection device and an external device. .

[0007]

When the cadide compound is irradiated with light to release the reaction product, the amount of emission is proportional to the amount of light irradiation. The wavelength of the dissociated irradiation light differs depending on the cadide compound. When there are two or more kinds of reactants, the reaction can be selectively started by selecting the wavelength of the excitation light to be irradiated by using each of the reactants as a cadide compound that dissociates at a different wavelength. The cadide compound is a substance which is made to release a substance involved in the reaction by dissociation by light irradiation. Such a caged compound

[0008]

[Chemical 1]

It is a compound having a structure such as Here, R1 to R5 are arbitrary functional groups, and the substances involved in the reaction are included therein. Such a caged compound is
A compound obtained by chemically modifying a substance involved in a reaction is reacted with a compound forming the skeleton of a cadide compound, and is dissociated by irradiation with light in the ultraviolet or visible region to release the substance involved in the reaction.

[0010]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows the basic structure. Two
Is a light source unit, 4 is a light source controller that controls the irradiation power, irradiation timing, and irradiation time of the light source unit 2, and 6 is the wavelength of the light emitted from the light source unit 2 to the sample 10 containing the caged compound. It is a wavelength selecting device for selecting. The control interface 8 exchanges signals between the light source controller 4 and the wavelength selection device 6 and the main body of the external device.

A flash lamp or a laser device can be used as the light source 2. The wavelength selection device 6 is provided with a molochromator or a plurality of optical filters, and the transmitted light is selected so as to have an appropriate distribution in the selected monochromatic light or the selected wavelength range. The control interface 8 is connected to the main body, the light source controller 4 and the wavelength selecting device 6, and controls the light source 2 and the wavelength selecting device 6 by software of the main body. In FIG. 1, illustration of optical elements such as a lens, a diaphragm, and a shutter is omitted.

FIG. 2 shows an example of controlling the irradiation amount. The irradiation dose is the product of irradiation power and irradiation time per unit time. As shown in FIG. 2A, the irradiation power is controlled so as to increase proportionally with time, and the irradiation power is controlled so as to be intermittently irradiated at fixed time intervals. The irradiation amount at this time changes with time as shown in FIG. The cadide compound dissociates in proportion to the irradiation dose and the reactant is released. By changing the irradiation profile, the irradiation amount, that is, the emission amount of the reactant can be controlled freely and in real time.

FIG. 3 shows an example in which a plurality of reactants are contained in the reaction system as cadide compounds, and these cadide compounds are dissociated by light having different wavelengths to release the reactants. . Here, two kinds of reactants are added to the reaction system as the respective cadide compounds A and B. It is assumed that the cadide compound A dissociates at the wavelength λa to release the reactant A ′, and the cadide compound B dissociates at the wavelength λb to release the reactant B ′. Figure 3 now
Assuming that light irradiation is performed with the irradiation profile shown in (A), the reaction substance A ′ shows a change in the emission amount as shown in (B), and the reaction substance B ′ has a change in (C). Shows the change in release as shown.

When the reaction substance is released from the cadide compound by light irradiation and the reaction is measured by a fluorescence microscope or the like, the measurement is performed during the irradiation of the excitation light (light source trigger) as shown in FIG. The reaction system can be observed while controlling the timing and controlling the release amount. As the irradiation amount control of the light source trigger for releasing the reactant, various irradiation profiles are used in addition to those shown in FIGS. 2 and 3, and various irradiation profiles are combined by combining the reactant release with the irradiation profile and the measurement timing. The reaction of the conditions can be measured.

FIG. 5 shows an embodiment in which the present invention is applied to a fluorescence microscope. In the fluorescence microscope, the sample 10 is placed on the sample table, observation light is emitted from the lower part of the sample table by the observation light source 20, and the sample is condensed by the condenser lens 22 on the sample 10. The observation light transmitted through the sample 10 is collected by the objective lens 24, and the fluorescence is selectively detected by the detector 26.

A photoexcitation device is provided to excite the cadide compound contained in the sample 10 to release the reaction substance. The photoexcitation device is provided with a light source controller 4 and a light source 2 whose irradiation power is controlled by the light source controller 4. ,
A condenser lens 12 for condensing the light from the light source 2 and a wavelength selecting device 6 for guiding the light condensed by the condensing lens 12 are provided, and a shutter is provided between the condensing lens 12 and the wavelength selecting device 6. The irradiation timing and the irradiation time are controlled by the shutter 14. The slit 16 changes the irradiation light into a light beam having an appropriate size. Light source 2,
The lens 12, the shutter 14 and the slit 16 form a light source unit. The sample 10 is irradiated with the irradiation light of which the wavelength is selected by the wavelength selection device 6 by the condenser lens 18.

The main controller 30 controls the light applied to the sample through the control interface 8 and takes in the detection signal from the detector 26 of the fluorescence microscope. The main controller 30 corresponds to an external device when viewed from the photoexcitation device of the present invention. The present invention can be applied not only to the fluorescence microscope but also to various optical measuring devices such as a spectrofluorometer.

[0018]

INDUSTRIAL APPLICABILITY In the present invention, a light source controller for controlling the irradiation power, the irradiation timing and the irradiation time of a light source for irradiating a cadide compound to release a reaction substance is provided, and a signal is exchanged between the light source controller and an external device. Since the light source is controlled while performing the above, it becomes possible to control the amount of the released substance causing the reaction in real time. By combining with a wavelength selection device, it becomes possible to control the emission amount of a plurality of reactants in real time. By combining with a fluorescence microscope or a spectrofluorescence system, it becomes possible to perform light source control or wavelength control in conjunction with measurement by those optical instruments.

[Brief description of drawings]

FIG. 1 is a block diagram showing an embodiment.

FIG. 2 is a diagram showing an example of release amount control.

FIG. 3 is a diagram showing another example of release amount control.

FIG. 4 is a timing diagram showing the timing of excitation of a cadide compound and observation with an optical instrument.

FIG. 5 is a schematic configuration diagram showing an example in which the present invention is applied to a fluorescence microscope.

[Explanation of symbols]

 2 light source 4 light source controller 6 wavelength selection device 8 control interface 10 sample

Claims (2)

[Claims] 1. An apparatus for irradiating a caged compound installed in an optical measuring apparatus with excitation light, a light source section, a light source controller for controlling irradiation power, irradiation timing and irradiation time of the light source section, and a light source thereof. An optical excitation device, comprising: a control interface for exchanging signals between a controller and an external device. 2. The apparatus further comprises a wavelength selection device that selects a wavelength of light emitted from the light source unit to irradiate a caded compound, and the control interface outputs a signal between the wavelength selection device and an external device. The photoexcitation device according to claim 1, wherein the photoexcitation device transmits and receives.