JP3158603U - Spectrofluorometer - Google Patents

Abstract

[PROBLEMS] To measure a spectrofluorometer, for example, may be a sample that belongs to a biological system and it is difficult to obtain a large amount of sample, or a minute sample because it is expensive. However, the present invention provides a fluorescence spectrophotometer capable of highly sensitive fluorescence measurement capable of performing these measurements. By using a test tube-shaped thin tube cell as a sample cell, the amount of sample to be measured can be reduced, and the sample cell can be irradiated by using the test tube-shaped thin tube cell. In order to prevent a decrease in the excitation light, the condenser lens 3 is disposed on the optical path of the excitation light. [Selection] Figure 1

Description

  The present invention relates to a spectrofluorometer, and more particularly to a spectrofluorometer suitable for measuring fluorescence of trace substances in a solution.

  In general, a spectrofluorometer is provided with a light source 10 that generates excitation light, a spectroscope (excitation-side concave diffraction grating) 11 that separates light from the light source 10 for each wavelength, and wavelength dispersion by the spectroscope 11 as shown in FIG. Excitation-side slit 14 for narrowing down the excitation light, cell holder 21 for accommodating the sample to be measured, spectrometer (fluorescence-side concave diffraction grating) 12 for wavelength-dispersing the fluorescence emitted from the sample, and wavelength dispersion by the spectrometer 12 It comprises a fluorescence side slit 15 and a detector 13 for narrowing down the emitted fluorescence.

  Light emitted from the light source 10 is wavelength-dispersed by the excitation-side concave diffraction grating 11, passes through the excitation-side slit 14, and is irradiated to the square cuvette cell 18 inserted in the cell holder 21. The fluorescence emitted from the sample is also wavelength-dispersed by the fluorescent concave grating 12 and passes through the fluorescent slit 15 and then detected by the detector 13 (see Patent Document 1).

  FIG. 5 shows a configuration around a sample cell in a general spectrofluorometer. When fluorescence measurement is performed on a liquid sample, a square cuvette cell 18 is generally used, and one made of glass or quartz is selected depending on the wavelength range to be measured.

  In order to reduce the amount of sample necessary for measurement as much as possible, as shown in FIG. 6, for example, as shown in FIG. In this case, the excitation light hits the inner wall of the cell to prevent the measurement data from becoming unstable. It is necessary to select the excitation side slit 20 having a small slit width.

  In FIG. 5, the excitation light passes through the excitation light path 22 of the cell holder 21 and is irradiated to the square cuvette cell 18 inserted in the cell holder 21. Fluorescence emitted from the sample passes through the fluorescence path 9 of the cell holder 21, passes through the fluorescence side slit (not shown), and is then detected by a detector (not shown). In FIG. 6, parts denoted by the same reference numerals as in FIG. 5 are the same parts as in FIG.

Japanese National Patent Publication No. 8-500183

  As a field of application of a spectrofluorometer, for example, it belongs to a biological system and it is difficult to take a large amount of sample, or it is expensive, so that the amount of sample required for fluorescence measurement is smaller. An analyzer is required. Further, as described above, since the necessary sample amount for fluorescence measurement is reduced, if the slit width to be set is limited to a small one, the fluorescence intensity itself becomes small and the measurement sensitivity is lowered.

  In order to solve the above-mentioned problems, the present invention provides a light source that emits excitation light, an excitation-side spectrometer that separates the excitation light, a sample cell that houses a sample to be measured, and a fluorescence that separates fluorescence emitted by the sample. In a spectrofluorometer comprising a side spectroscope and a detector for detecting fluorescence dispersed by the fluorescence side spectroscope, the sample cell is constituted by a test tube-shaped thin tube cell, and the light source A condensing lens is installed on the excitation light side optical axis between the test tube type narrow tube cells. Further, a concave reflecting mirror is installed on the opposite side of the excitation light incident side across the test tube-shaped thin tube cell, and emission of fluorescence generated from the sample is sandwiched between the test tube-shaped thin tube cell. A plane reflecting mirror may be installed on the opposite side.

  First, by using a test tube type thin tube cell as the sample cell, the amount of sample to be measured can be reduced, and the slit width to be set is reduced because the cell width is narrowed. Second, although the fluorescence intensity itself tends to decrease and the measurement sensitivity tends to decrease, the measurement sensitivity can be reduced by installing a condenser lens on the excitation light side optical axis between the light source and the test tube-shaped thin tube cell. Decline can be prevented.

  Third, an excitation light reflecting concave mirror is placed on the opposite side of the excitation light incident side across the test tube-shaped thin tube cell, and fluorescence generated from the sample is sandwiched between the test tube-shaped thin tube cell. By installing a fluorescence reflecting plane mirror on the opposite side of the emission side, the excitation light that has passed through the test tube-shaped thin tube cell is reflected by the excitation light reflecting concave mirror, and the sample in the test tube-shaped thin tube cell is again applied. Excitation provides a repeated excitation effect.

  In addition, since the fluorescence is emitted in all directions, the fluorescence emitted in the direction of the fluorescence reflecting plane mirror is reflected and taken into the detector side, and an effect of increasing the fluorescence intensity is obtained as compared with the state without these reflecting mirrors. Fourth, highly sensitive fluorescence measurement is possible even with a smaller amount of sample.

  Fifth, since the inside of the test tube type narrow tube cell has no corners and the tip of the inner surface of the cell has a spherical structure, the sample hardly remains, and the cell can be cleaned with a tool such as a brush. The cell can be reused. Therefore, it is possible to reduce the cost of sample preparation, and it is possible to reduce unnecessary labor of the operator's cleaning process.

It is a figure which shows this invention Example 1, and is a longitudinal cross-sectional view of a cell holder. It is a figure which shows this invention Example 1, and is a cross-sectional view of a cell holder. It is a figure which shows the optical layout of the spectrofluorometer of this invention Example 1. FIG. It is a figure which shows this invention Example 2, and is a cross-sectional view of a cell holder. It is a figure which shows the conventional Example, and is a cross-sectional view of the cell holder using a square cuvette cell. It is a figure which shows the prior art example, and is a cross-sectional view of a cell holder using an inner product reduced cuvette cell. It is a figure which shows the optical layout of the conventional spectrofluorometer.

  A sample tube-type thin tube cell is used as a sample cell, and a condensing lens is installed on the optical path of the excitation light in order to prevent a decrease in irradiation excitation light to the sample cell due to the use of the thin tube cell. To do.

  FIG. 3 is a diagram schematically showing the overall configuration of the spectrofluorometer. In the figure, the light emitted from the light source 10 is wavelength-dispersed by the excitation-side concave diffraction grating 11, passes through the excitation-side slit 14, passes through the condenser lens 3, and is applied to the test tube-shaped narrow tube cell 1. The The fluorescence emitted from the sample is also wavelength-dispersed by the fluorescent-side concave diffraction grating 12 and then detected by the detector 13 after passing through the fluorescent-side slit 15.

  1, 2 and 3 show a specific example 1. FIG. As shown in FIGS. 1 and 2, a test tube-shaped thin tube cell 1 is used as a sample cell, and the test tube-shaped thin tube cell 1 is inserted through the tube cell insertion path 4 and attached to the cell holder 2. The cell holder 2 is arranged in the optical layout of the spectrofluorometer of the first embodiment according to the present invention shown in FIG.

  The excitation light passes through the condenser lens 3, passes through the excitation light path 5 of the cell holder 2, and is irradiated to the test tube-shaped thin tube cell 1 inserted into the cell holder 2. Fluorescence emitted from the sample passes through the fluorescence path 6 of the cell holder 2, passes through the fluorescence side concave diffraction grating 12, is similarly wavelength dispersed, passes through the fluorescence side slit 15, and is detected by the detector 13.

  Since the test tube-shaped thin tube cell 1 is used, it is impossible to irradiate the same amount of excitation light as that of the sample of the conventional wide square cuvette cell 18, and therefore the test tube is narrowed down. The condensing lens 3 is installed at the entrance of the excitation light path 5 so that the same amount of excitation light can be irradiated onto the narrow tube cell 1. The condensing lens 3 may be a spherical lens, an aspherical lens, or any other type. Examples of dimensions of the test tube-shaped thin tube cell 1 include a cell having a diameter of 2 mm or 3 mm.

  Example 2 is shown in FIG. An excitation light reflecting concave mirror 7 is installed on the side where the excitation light passes through the test tube-shaped narrow tube cell 1, and a fluorescence reflecting plane mirror 8 is installed in the opening 17 of the fluorescence path 16 on the opposite side of the fluorescence emission side. By installing the excitation light reflecting concave mirror 7, the excitation light that has passed through the test tube thin tube cell 1 is reflected on the excitation light reflecting concave mirror 7 to excite the sample in the test tube thin tube cell 1 again. Thus, a repeated excitation effect can be obtained.

  Further, by the installation of the fluorescence reflecting plane mirror 8, the fluorescence is emitted in all directions, and the fluorescence emitted in the direction of the fluorescence reflecting plane mirror 8 is reflected and taken into the detector side. Accordingly, it is possible to increase the fluorescence intensity as compared with the state without the reflecting mirror. In FIG. 4, parts denoted by the same reference numerals as in FIG. 2 are the same parts as in FIG.

DESCRIPTION OF SYMBOLS 1 Test tube type thin tube cell 2 Cell holder 3 Condensing lens 4 Tube cell insertion path 5 Excitation light path 6 Fluorescence path 7 Excitation light reflection concave mirror 8 Fluorescence reflection plane mirror 9 Fluorescence path 10 Light source 11 Excitation side concave diffraction grating 12 Fluorescence side concave surface Diffraction grating 13 Detector 14 Excitation side slit 15 Fluorescence side slit 16 Fluorescence path 17 Opening 18 Square cuvette cell 19 Inner product reduction type cuvette cell 20 Excitation side slit 21 Cell holder 22 Excitation light path

Claims (2)

  A light source that emits excitation light, an excitation-side spectrometer that separates the excitation light, a sample cell that houses a sample to be measured, a fluorescence-side spectrometer that separates fluorescence emitted by the sample, and a fluorescence-side spectrometer In a spectrofluorometer equipped with a detector for detecting spectroscopic fluorescence, the sample cell is constituted by a test tube type narrow tube cell, and excitation light between the light source and the test tube type thin tube cell A spectrofluorometer characterized in that a condensing lens is installed on the side optical axis.   A concave reflecting mirror is installed on the opposite side of the excitation light incident side across the test tube-shaped thin tube cell, and on the emission side of the fluorescence generated from the sample across the test tube-shaped thin tube cell. 2. The spectrofluorometer according to claim 1, wherein a plane reflecting mirror is installed on the opposite side.

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