JPS60162942A - Spectral fluorophotometer - Google Patents

Abstract

PURPOSE:To enable correction over a wide wavelength region by determining the specific 1st and 2nd correction functions with respect to different wavelength regions and correcting spectral data in the stage of measurement with the correction function in the wide wavelength region determined from at least one thereof. CONSTITUTION:The data from respective plates 4, 5 where a photon meter and tungsten lamp (not shown in a figure) are installed in a sample chamber 3 are read via an A/D converter 9 into a RAM16 in correction of the wavelength characteristics of a fluorescent side optical system consisting of a spectroscope 6 and detector 7 for detecting the fluorescence of the sample (not shown in the figure) in the chamber 3. Storage parts 18, 19 for respective fluorescent side correction functions by photon meter method and lamp method are further provided to the RAM16. The characteristic in a 200-600mm. wavelength region is measured by photon meter method and the correction function thereof is stored in the part 18. The correction function in a 500-800mm. wavelength region by lamp method is stored in the part 19. The entire part of the correction function by lamp method is multiplied by a factor so that the correction functions for 500mm. by both methods coincide with each other.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Application of the Invention] The present invention relates to a spectrofluorophotometer, and more particularly to a spectrofluorophotometer that is capable of recording a true spectrum removed over a wide wavelength range of the spectrum.

[Background of the invention]

The commonly performed spectral correction of a fluorophotometer is (
Fluorescence analysis - Kodansha Scientific (P81-8
As shown in 7), the device function on the excitation side is read using a photon meter (usually high concentration rhodamine B as a standard sample), and the fluorescence side is measured using a photon meter. This is a method of calculating an apparatus function (correction function) using an excitation-side apparatus function based on the amount of light. As shown in the literature, there is also a method (2) in which the device function on the fluorescent side is read using a standard tungsten lamp whose output is known. Regarding spectral correction on the fluorescent side, the former method is limited to a wavelength range of 600 discs from 2001TIn, and the latter method is limited to a wavelength range of 330I and above, so there is no effective correction means for all necessary wavelength ranges. did not previously exist.

[Purpose of the invention]

The object of the present invention is to provide two
It is an object of the present invention to provide a spectrofluorophotometer that can freely record continuous corrected spectra in the wavelength range from around 00 nits to around 800 Ilwns.

[Summary of the invention]

In the present invention, prior to sample measurement, excitation light is irradiated onto a standard sample, and the first correction function of the spectrum obtained based on the amount of excitation light measured by a photon meter and the amount of light irradiated from a light source with known output are A second correction function of the spectrum measured by the system and determined from the ratio of the measured value and the theoretical value is set for each different wavelength range, and one of these correction functions is adjusted so that the two correction functions are continuous. Alternatively, the present invention is characterized in that a corrective function (3) in a wide wavelength range is calculated by correcting both of them, and the spectral data is corrected based on the correction function data at the time of sample measurement.

[Embodiments of the invention]

Embodiments of the present invention will be described based on the drawings.

FIG. 1 shows a principle diagram of an embodiment of the present invention. The wavelength characteristics of the device on the fluorescence side are determined by the optical quantum meter method and the tungsten lamp method, and the correction function curves, which are the reciprocals thereof, are connected at 500I. The first step is
The wavelength characteristics of the device in the wavelength range of 200I to 600nwn are measured using a photon meter, and the correction function, which is the reciprocal thereof, is stored in a storage device. Next, when reading the correction function in the wavelength range of 500 ntn or more into the storage device by the tungsten lamp method, the tungsten lamp is The entire correction value of the method is multiplied by a coefficient and stored in a storage device. The correction function thus obtained enables continuous (4) spectral correction in the wavelength range from 200+in to 800in.

FIG. 2 shows the configuration of an embodiment of the spectrofluorophotometer according to the present invention. In the figure, light emitted from the light source 1 is guided to the sample chamber 3 via the excitation side spectrometer 2. The sample is irradiated. Fluorescence emitted from the sample enters a detector 7 via a fluorescence-side spectrometer 6. What we are currently targeting is the correction of the wavelength characteristics of the fluorescent side optical system consisting of the fluorescent side spectrometer 6 and the detector 7.

In the sample chamber 3, there is a photon meter installation location 4, but apart from this, there is a tungsten lamp installation location 5. Both the wavelength characteristic data of the device and the actual measurement data are sent to the detector 7.
R, A via A/D converter 9 and pass line 12
Read into M16.

The RAM 16 includes an excitation side correction function storage section 17, a storage section 18 that stores a fluorescence side correction function based on the photoelectron method, and a storage section 19 that stores a fluorescence side correction function based on the tungsten lamp method.
are set up independently. The microprocessor unit (MPU) 14 calculates the above-mentioned correction function based on the program stored in the R,0M15, and stores the correction function data (5) in the RAM 16. The measurement data of the sample is corrected based on the corrected correction function data, and the corrected spectrum is recorded in the recording system 11 via the D/A converter 10.

FIG. 3 is an example of a corrected spectrum actually obtained according to the above embodiment. Correction spectra covering a wide wavelength range have been recorded. This data is a corrected spectrum taken for the purpose of quantum yield calculation, and it is necessary to correctly correct the tail of the spectrum. This is one of the examples in which the present invention is most effectively utilized.

〔Effect of the invention〕

According to the present invention: 1) A fluorescent side correction spectrum can be obtained over a wavelength range from around 300 on to over 600 I.

2) Quantum yield calculation becomes easy.

3) There is no need to use an expensive standard tungsten lamp that can be used up to around 330 + nm;
It has the effect that 0 mn+ or less is sufficient.

(6)

[Brief explanation of the drawing]

FIG. 1 is a diagram for explaining the present invention in detail, FIG. 2 is a block diagram of an embodiment of the spectrofluorophotometer according to the present invention, and FIG. 3 is a diagram for explaining the spectrofluorophotometer according to the present invention. It is a figure which shows an example of the obtained correction spectrum. DESCRIPTION OF SYMBOLS 1... Light source, 2... Excitation side spectrometer, 3... Sample chamber, 6... Fluorescence side spectrometer, 7... Detector, 8... Mechanical system control circuit, 9... A/I) converter, 10...D/
A conversion fee, 14...MPU, +5・ROM, 16...
・RAM. Agent Patent Attorney Akio Takahashi (7)

Claims (1)

[Claims] (1) In a spectrofluorophotometer that irradiates a sample with excitation light from a light source and analyzes fluorescence emitted from the sample, a photon meter and an output are provided in a sample chamber in which the sample is installed. A known light source, a first correction function of spectrum 1H obtained based on the amount of excitation light measured by the photon meter by irradiating the standard sample with excitation light prior to 1q constant on the sample, and irradiation from the light source with the known output. A second correction function of the spectrum calculated from the ratio of the measured value and the theoretical value is determined for each different wavelength range, so that the two correction functions are continuous. It has an arithmetic device that performs a correction operation on one or both of these correction functions to obtain one correction function in a wide wavelength range, and a storage device that stores the correction function data determined by the arithmetic device, In particular, the arithmetic device corrects the spectral data based on the correction function data stored in the storage device at the time of sample measurement (1).
) A spectrofluorophotometer. 2. The spectrofluorophotometer according to claim 1, wherein the light source with a known output is a tungsten lamp.