JP6920943B2 - Optical measuring device and optical measuring method - Google Patents

Description

The present invention relates to an apparatus and a method for measuring the intensity of light generated in a sample in response to irradiation of the sample with light.

The sample can be analyzed by measuring the intensity of the light generated in the sample in response to the irradiation of the sample with light. Devices that perform such light measurement include a light source that outputs the irradiation light that is applied to the sample, a photodetector that detects the intensity of the generated light generated in the sample and outputs a detection signal, and the detection signal. A processing unit that outputs a digital value according to a value (analog value) is provided (see Patent Document 1).

The light generated in the sample in response to the light irradiation is fluorescence, scattered light, or light generated by a nonlinear optical phenomenon (for example, harmonic light or Raman scattered light). By detecting the fluorescence intensity, the concentration of the fluorescent substance contained in the sample can be measured. By detecting the intensity of scattered light, it is possible to measure the concentration and size of scattered substances contained in the sample. Further, the non-linearity of the sample can be measured by detecting the intensity of the light generated by the non-linear optical phenomenon.

Japanese Unexamined Patent Publication No. 2004-257901

When measuring the intensity of light generated in a sample when the sample is irradiated with irradiation light having a plurality of peak wavelengths, the device including the above-mentioned light source, photodetector and processing unit is used to obtain the number of peak wavelengths of the irradiation light. Only need to be prepared.

The present invention has been made to solve the above problems, and more easily, it is possible to measure the intensity of light generated in a sample when the sample is irradiated with irradiation light having a plurality of peak wavelengths. It is an object of the present invention to provide a device and a method capable of providing the same.

The optical measuring device of the present invention is a device that measures the intensity of light generated in a sample in response to light irradiation of the sample, and (1) any one peak wavelength selected from a plurality of peak wavelengths. An irradiation unit that includes a light source that outputs the irradiation light of the above and irradiates the sample with the irradiation light of each peak wavelength output from the light source over different irradiation periods, and (2) one-to-one correspondence with multiple peak wavelengths Each light detector radiates the generated light generated in the sample in a direction different from the irradiation direction of the irradiation light in response to the irradiation of the irradiation light of the corresponding peak wavelength. The detection unit that receives the generated light, detects the intensity of the received generated light and outputs the detection signal, and (3) the detection signals output from each of the multiple optical detectors are input in common as analog values. An analog value input to the input terminal is input during the irradiation period when the light source outputs the irradiation light of each peak wavelength, and the digital value corresponding to this analog value is set to the peak wavelength. It is provided with a processing unit that outputs a value indicating the intensity of the generated light generated in the sample in response to the irradiation of the irradiation light.

In one aspect of the optical measuring device of the present invention, the irradiation unit outputs irradiation light of each peak wavelength from the light source over different irradiation periods based on a synchronization signal indicating the irradiation period of the irradiation light of each peak wavelength. It is preferable that the processing unit performs conversion processing from an analog value to a digital value based on the synchronization signal. Further, it is preferable that the optical measuring device further includes a control unit that outputs a synchronization signal to the irradiation unit and the processing unit.

In one aspect of the optical measuring device of the present invention, the processing unit outputs a signal converter that outputs a voltage value corresponding to the value of the detection signal input to the input terminal, and a digital value of the voltage value output from the signal converter. It is preferable to include an AD converter that converts to and outputs the digital value. The signal converter preferably includes an integrator that has a capacitance unit that stores electric charges based on a detection signal input to the input terminal and outputs a voltage value corresponding to the amount of accumulated electric charges. Further, the signal converter includes the integrator which is the first integrator and the integrator which is the second integrator, and the first integrator and the second integrator include irradiation light having a plurality of peak wavelengths. It is preferable to alternately input the detection signals in synchronization with each irradiation period.

In one aspect of the optical measuring device of the present invention, the irradiation unit includes a light source having a plurality of light emitting elements having different peak wavelengths of output light, and irradiates from any one light emitting element selected from the plurality of light emitting elements. It is preferable to output light and irradiate the sample with the irradiation light. Further, the irradiation unit includes a light source having a variable output light wavelength, outputs irradiation light of any one peak wavelength selected from a plurality of peak wavelengths from the light source, and irradiates the sample with the irradiation light. Is also suitable.

The light measurement method of the present invention is a method of measuring the intensity of light generated in a sample in response to light irradiation of the sample, and (1) any one peak wavelength selected from a plurality of peak wavelengths. In the irradiation unit including the light source that outputs the irradiation light of, the light irradiation step of irradiating the sample with the irradiation light of each peak wavelength output from the light source over different irradiation periods, and (2) for a plurality of peak wavelengths. In the detection unit including a plurality of light detectors provided in a one-to-one correspondence, each light detector irradiates the irradiation light among the generated lights generated in the sample in response to the irradiation of the irradiation light of the corresponding peak wavelength. A light detection step that receives generated light emitted in a direction different from the direction, detects the intensity of the received generated light, and outputs a detection signal, and (3) detection output from each of a plurality of optical detectors. In a processing unit having an input terminal in which signals are commonly input as analog values, the analog value input to the input terminal is input during the irradiation period when the light source outputs the irradiation light of each peak wavelength, and this analog value is used. It includes a signal processing step of outputting the corresponding digital value as a value representing the intensity of the generated light generated in the sample in response to the irradiation of the irradiation light of the peak wavelength.

In one aspect of the light measurement method of the present invention, in the light irradiation step, irradiation light of each peak wavelength is output from the light source over different irradiation periods based on a synchronization signal indicating the irradiation period of the irradiation light of each peak wavelength. However, in the signal processing step, it is preferable to perform conversion processing from an analog value to a digital value based on the synchronization signal. Further, the optical measurement method preferably further includes a synchronization step of outputting a synchronization signal to the irradiation unit and the processing unit.

In one aspect of the optical measurement method of the present invention, the signal processing step includes a signal conversion step that outputs a voltage value corresponding to the value of the detection signal input to the input terminal, and a signal conversion step that converts the output voltage value into a digital value. It is preferable to include an AD conversion step for outputting the digital value. In the signal conversion step, it is preferable to use an integrator to accumulate electric charges based on the detection signal input to the input terminal and output a voltage value corresponding to the amount of the accumulated electric charges. Further, in the signal conversion step, it is preferable to use two integrators to alternately accumulate charges in synchronization with the irradiation period of each irradiation light having a plurality of peak wavelengths.

In one aspect of the light measurement method of the present invention, in the light irradiation step, a light source having a plurality of light emitting elements having different peak wavelengths of output light is used, and from any one light emitting element selected from the plurality of light emitting elements. It is preferable to output the irradiation light and irradiate the sample with the irradiation light. Further, in the light irradiation step, a light source having a variable output light wavelength is used, irradiation light of any one peak wavelength selected from a plurality of peak wavelengths is output from the light source, and the irradiation light is irradiated to the sample. It is also preferable to do so.

The generated light may be fluorescence, scattered light, or light generated by a nonlinear optical phenomenon.

As another aspect, the light measuring device of the present invention is a device that measures the intensity of light generated in a sample in response to irradiation of the sample placed in the sample container, and (1) faces each other. Between the first and second surfaces, a third surface different from these, a first through hole provided between the first surface and the second surface, and between the third surface and the first through hole. It has a second through hole provided, and a sample container is inserted into the first through hole from the side of the first surface, and the sample in the inserted sample container is at a position facing the second through hole. In addition to positioning the sample container in this way, a metal block in which the inner wall surface of the first through hole is in thermal contact with the sample container, and (2) provided on the side of the second surface of the metal block, are provided in the first through hole of the metal block. A light source that outputs the irradiation light emitted to the sample in the inserted and positioned sample container into the first through hole, and (3) the first metal block provided on the third surface side of the metal block. It has a light receiving surface that receives the generated light generated in response to the irradiation of the irradiation light in the sample in the sample container that is inserted into the through hole and positioned, and the generated light that reaches the light receiving surface through the second through hole. It includes an optical detector that detects the intensity and outputs a detection signal. The metal block has a plurality of third surfaces, has a plurality of second through holes provided between any one of the plurality of third surfaces and the first through hole, and has a plurality of light sources. The irradiation light of any one of the peak wavelengths selected from the above peak wavelengths is output, and the photodetector is provided in any of the plurality of second through holes in a one-to-one correspondence with respect to the plurality of peak wavelengths. Has been done.

According to the present invention, it is possible to more easily measure the intensity of light generated in a sample when the sample is irradiated with irradiation light having a plurality of peak wavelengths.

FIG. 1 is a diagram showing an overall configuration of the optical measuring device 1 of the present embodiment. FIG. 2 is a diagram showing a main configuration of the optical measuring device 1 of the present embodiment. FIG. 3 is a timing chart showing an operation example of the optical measuring device 1 of the present embodiment. FIG. 4 is a timing chart showing another operation example of the optical measuring device 1 of the present embodiment. FIG. 5 is a diagram showing another configuration example of the signal converter 31 of the processing unit 30. FIG. 6 is a diagram showing still another configuration example of the signal converter 31 of the processing unit 30. FIG. 7 is a timing chart showing an operation example of the optical measuring device 1 when the signal converter 31B is used. FIG. 8 is a diagram showing another configuration example of the processing unit 30. FIG. 9 is a timing chart showing an operation example of the optical measuring device 1 when the processing unit 30A is used. FIG. 10 is a diagram showing a configuration of a modified example of the measurement optical system. FIG. 11 is a vertical cross-sectional view showing the configuration of a preferred example of the measurement optical system. FIG. 12 is a vertical cross-sectional view showing the configuration of the metal block 50 in the measurement optical system shown in FIG. FIG. 13 is a vertical cross-sectional view showing the configuration of the optical element holding portion 60 in the measurement optical system shown in FIG. FIG. 14 is a cross-sectional view of the measurement optical system shown in FIG. FIG. 15 is a vertical cross-sectional view showing the configuration of the first modification of the measurement optical system. FIG. 16 is a vertical cross-sectional view showing the configuration of a second modification of the measurement optical system. FIG. 17 is a perspective view showing a first configuration example of the light source 11. FIG. 18 is a cross-sectional view showing a second configuration example of the light source 11. FIG. 19 is a cross-sectional view showing a third configuration example of the light source 11. FIG. 20 is a perspective view showing the filter wheel 96 used in the third configuration example of the light source 11.

Hereinafter, embodiments for carrying out the present invention will be described in detail with reference to the accompanying drawings. In the description of the drawings, the same elements are designated by the same reference numerals, and duplicate description will be omitted. The present invention is not limited to these examples, and is indicated by the scope of claims, and is intended to include all modifications within the meaning and scope equivalent to the scope of claims.

The optical measuring device or the optical measuring method of the present invention measures the intensity of light generated in a sample in response to irradiation of irradiation light having a plurality of peak wavelengths. In the following embodiments, the number of peak wavelengths is determined. It will be described as 3. N appearing below is each integer of 1 or more and 3 or less. Further, the light measuring device or the light measuring method of the present invention measures the intensity of light (fluorescence, scattered light, or light generated by a nonlinear optical phenomenon) generated in a sample irradiated with light. In the following embodiment, the case of measuring the fluorescence intensity will be described. Further, it is assumed that the sample contains three kinds of fluorescent substances.

FIG. 1 is a diagram showing an overall configuration of the optical measuring device 1 of the present embodiment. FIG. 2 is a diagram showing a main configuration of the optical measuring device 1 of the present embodiment. The optical measuring device 1 measures the intensity of fluorescence generated by irradiating the sample placed in the sample container 2 with excitation light, and measures the irradiation unit 10, the detection unit 20, the processing unit 30, the control unit 40, and the input. A unit 41, a display unit 42, and a storage unit 43 are provided.

The irradiation unit 10 includes a light source 11, an excitation light transmission filter 12, and a light source drive circuit 13. The light source 11 outputs irradiation light (excitation light) having any one peak wavelength λ _{n selected from the three peak wavelengths λ 1 to λ 3.} The irradiation light having a peak wavelength λ _{n (hereinafter, may be referred to as “irradiation light λ n} ”) can selectively excite the nth fluorescent substance among the three types of fluorescent substances contained in the sample. A lens for increasing the directivity of light may be provided on the light output side of the light source 11. This lens may be integrated with the light source 11 or may be separate from the light source 11.

The excitation light transmission filter 12 is optically connected to the light source 11 and inputs the light output from the light source 11. The excitation light transmission filter 12 _{selectively transmits the irradiation lights λ 1 to λ 3} that excite the fluorescent substance contained in the sample, and outputs the irradiation light to the sample container 2. The transmission wavelength of the excitation light transmission filter 12, that is, the excitation light wavelength is appropriately selected according to the fluorescent substance contained in the sample placed in the sample container 2. The excitation light transmission filter 12 may be a dielectric multilayer filter having good filter characteristics, or may be a colored glass filter. The excitation light transmission filter 12 may block light having a wavelength of detected generated light (fluorescence). A specific configuration example of the light source 11 will be described later.

The light source drive circuit 13 is electrically connected to the light source 11 and drives the light source 11 to emit light. The light source 11 is driven by the light source drive circuit 13 instructed by the control unit 40, and outputs _{irradiation lights λ 1 to λ 3 to be irradiated to the sample container 2 over different irradiation periods.} That is, the light source 11 does not output the irradiation light of the other peak wavelength during the period of outputting the irradiation light of a certain peak wavelength among the _{irradiation lights λ 1 to λ 3.} The irradiation unit 10 irradiates the sample with irradiation light of each peak wavelength output from the light source 11 over different irradiation periods.

Detector 20 includes an optical detector ₂₁ 1 to 21 ₃ and the fluorescence transmitting filter ₂₂ 1-22 ₃ provided to the irradiation light lambda ₁ to [lambda] ₃ in a one-to-one correspondence. For convenience of illustration, it does not show the fluorescence transmitting filter ₂₂ 1-22 ₃ in FIG.

Each fluorescence transmission filter 22 _n selectively transmits the fluorescence (generated light) generated in the sample according _{to the irradiation of the irradiation light λ n} to the sample placed in the sample container 2, and corresponds to the fluorescence. Output to the photodetector 21 _n. Each fluorescence transmission filter 22 _n blocks the fluorescent light other than (e.g., light transmitted through the excitation light transmission filter 12). Transmission wavelength of each fluorescent transmission filter 22 _n is selected appropriately depending on the fluorescent material of the first n among the three types of fluorescent substance contained in the sample placed in the sample container 2. Each fluorescence transmission filter 22 _n is to filter characteristic may be a good dielectric multilayer filter may be a colored glass filter.

Each photodetector 21 _n is optically connected to a corresponding fluorescence transmission filter 22 _n and receives fluorescence transmitted through the fluorescence _{transmission filter 22 n.} Each photodetector 21 _n detects the intensity of the fluorescence received, and outputs a detection signal indicating the fluorescence intensity to the processing unit 30. Each photodetector 21 _n is, for example, a photodiode, an avalanche photodiode, a photomultiplier tube, or the like. Detector 20 by the photodetectors 21 _n, by receiving the generated light that is emitted in a direction different from the irradiation direction of the irradiation light of the generated light generated in the sample in response to irradiation of the corresponding illumination light lambda _n , Detects the intensity of the received generated light and outputs a detection signal.

Light source 11, the excitation light transmitting filter 12, the sample container 2, the fluorescence transmitting filter ₂₂ 1-22 ₃ and light detector ₂₁ 1 to 21 ₃ measuring optical system including the inside of the dark box so as stray light from the outside does not enter Be placed.

Processor 30 includes an optical detector ₂₁ 1 to 21 ₃ and input terminals electrically connected. Its input terminal, an optical detector 21 ₁ to 21 ₃ detection signal output from each is input to the common as an analog value. The processing unit 30 inputs an analog value input to the input terminal during the irradiation period when the light source 11 of the irradiation unit 10 _{outputs the irradiation light λ n,} and the digital value corresponding to the analog value is set to the irradiation light λ _n. Is output to the control unit 40 as a value indicating the intensity of the generated light (fluorescence) generated in the sample irradiated with. The details of the processing unit 30 will be described later.

The control unit 40 is electrically connected to the light source drive circuit 13, and by controlling the light source drive circuit 13, each irradiation light λ _n is emitted from the light source 11 over a predetermined irradiation period. The control unit 40 is electrically connected to the processing unit 30, controls the operation of the processing unit 30, and inputs a digital value output from the processing unit 30 to control the intensity of fluorescence generated in each sample. Obtain and analyze the sample based on this fluorescence intensity. The control unit 40 stores these digital values, the fluorescence intensity, and the analysis result in the storage unit 43. The control unit 40 includes a CPU that performs the above control and analysis, and is, for example, a computer such as a personal computer, a smart device, a microcomputer, or a cloud server. The connection between the control unit 40 and other components may be wireless as well as wired, and may include a connection by network communication.

The control unit 40 generates a synchronization signal indicating the irradiation period of each irradiation light λ _n from the light source 11 of the irradiation unit 10, and outputs this synchronization signal to each of the irradiation unit 10 and the processing unit 30. _{The light source drive circuit 13 of the irradiation unit 10 outputs each irradiation light λ n} from the light source 11 over different irradiation periods based on the synchronization signal output from the control unit 40. The processing unit 30 performs conversion processing from an analog value to a digital value based on the synchronization signal output from the control unit 40. This synchronization signal may be a periodic clock signal or a trigger signal that directly indicates each operation timing. When the synchronization signal is a clock signal, each of the light source drive circuit 13 and the processing unit 30 counts the pulses of the clock signal and determines each operation timing in response to reaching a predetermined count value. The synchronization signal does not have to be output from the control unit 40, and one of the light source drive circuit 13 and the processing unit 30 may be generated and output to the other.

The input unit 41 is electrically connected to the control unit 40 and receives inputs such as measurement conditions, measurement start and measurement end. The input unit 41 is, for example, a keyboard, a mouse, a touch screen, or the like. The input unit 41 may transmit a simple on / off such as a momentary switch or an alternate switch to the control unit 40.

The display unit 42 is electrically connected to the control unit 40 and displays measurement conditions and measurement results. The display unit 42 is, for example, a liquid crystal display, an organic EL display, a touch screen, or the like. Further, the display unit 42 may display by turning on / off a light emitting element such as an LED.

The storage unit 43 is electrically connected to the control unit 40, and stores the fluorescence intensity and the analysis result output from the control unit 40. The storage unit 43 is, for example, an internal memory or an external storage.

Next, the configuration of a main part of the optical measuring device 1 of the present embodiment will be described with reference to FIG. Hereinafter, each photodetector 21 _n will be described as a photodiode. The cathode of each photodetector 21 _n is a ground potential, and the anode of _{each photodetector 21 n is common.} Each photodetector 21 _n generates a charge according to incidence of light, and outputs from the anode current corresponding to the electric charge as a detection signal to the processing unit 30. The amount of electric charge generated per unit time in each photodetector 21 _n is a current value (analog value) according to the intensity of the received light.

The processing unit 30 includes a signal converter 31 and an AD converter 33. Signal converter 31 comprises a photodetector ₂₁ 1 to 21 ₃ input terminal T to be respective anodes electrically connected. Its input terminal T, photodetector 21 ₁ to 21 ₃ current value output from the respective (detection signal) is input to the common as an analog value. The signal converter 31 inputs an analog value input to the input terminal T during the irradiation period in which the light source 11 of the irradiation unit 10 _{outputs each irradiation light λ n, and a voltage value corresponding to the input analog value.} Is output. The AD converter 33 is electrically connected to the signal converter 31 and inputs a voltage value output from the signal converter 31. The AD converter 33 converts the input voltage value (analog value) into a digital value, and represents the intensity of the generated light (fluorescence) generated in the sample irradiated with _{each irradiation light λ n output from the light source 11.} The digital value is output as a value.

As shown in FIG. 2, the signal converter 31 may have an integrator configuration including an amplifier 311 and a capacitance unit 312 and a switch 313. The amplifier 311 has an inverting input terminal, a non-inverting input terminal, and an output terminal. The non-inverting input terminal of the amplifier 311 is set to the ground potential. Inverting input terminal of the amplifier 311, the light detector ₂₁ 1 to 21 ₃ of the detecting unit 20 are electrically connected to respective anode and. The output terminal of the amplifier 311 is electrically connected to the sample hold circuit 331 of the AD converter 33. The capacitance unit 312 and the switch 313 are connected in parallel to each other and are provided between the inverting input terminal and the output terminal of the amplifier 311.

In this signal converter 31, when the switch 313 is in the ON state, the charge accumulation of the capacitance unit 312 is initialized, and the voltage value of the output terminal of the amplifier 311 is also initialized. When the switch 313 is in the off state, electric charges are accumulated in the capacitance unit 312 based on the detection signal input to the inverting input terminal of the amplifier 311, and the charge corresponds to the amount of the electric charges stored in the capacitance unit 312. The voltage value is output from the output terminal of the amplifier 311. The voltage value output from the amplifier 311 is held by the sample hold circuit 331 of the AD converter 33. The voltage value held by the sample hold circuit 331 is output to the AD converter 33 and converted into a digital signal by the AD converter 33. The sample hold circuit does not have to be built in the AD converter 33. In this case, the sample hold circuit is electrically connected to each of the output terminal of the amplifier 311 and the AD converter 33. High-sensitivity photodetection can be performed by appropriately setting the length of the charge accumulation period and the capacitance value of the capacitance section 312 in the signal converter 31 having such an integrator configuration.

Next, an operation example of the optical measuring device 1 of the present embodiment and the optical measuring method of the present embodiment will be described. In the light measurement method of the present embodiment, the intensity of fluorescence generated in the sample by irradiating the sample placed in the sample container 2 with each irradiation light λ _{n is measured by using the light measurement device 1.}

FIG. 3 is a timing chart showing an operation example of the optical measuring device 1 of the present embodiment. This operation is performed based on the synchronization signal output from the control unit 40 in the synchronization step. This figure shows, in order from the top, the irradiation light lambda ₁ to [lambda] ₃ respectively output from the light source 11, the photodetector ₂₁ 1 to 21 ₃ of each output, the output of the output and the sample and hold circuit 331 of the signal converter 31 It is shown. Further, although the operation for two cycles is shown in this figure, the same applies to the subsequent cycles.

In the light irradiation step, the _{light source 11 outputs the irradiation light λ 1 during the period T 11} (time t _{10 to t 12} ), and the light source 11 irradiates the irradiation light λ 1 during the period T ₁₂ (time t _{12 to t 14} ) following this period T _11. The light λ ₂ is output, and the light source 11 outputs the _{irradiation light λ 3} during the period T ₁₃ (time t _{14 to t 20 ) following this period T 12.} Further, the _{light source 11 outputs the irradiation light λ 1} during the period T ₂₁ (time t _{20 to t 22 ) following this period T 13} , and the light source during the period T ₂₂ (time t _{22 to t 24} ) following this period T _21. 11 outputs the irradiation light λ ₂ , and the light source 11 outputs the _{irradiation light λ 3} during the period T ₂₃ (time t _{24 to t 30} ) following this period T _22. The periods T ₁₁ , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T ₂₃ do not overlap each other. In this way, the light source 11 outputs the _{irradiation lights λ 1 to λ 3 over different irradiation periods.}

In light detecting step, a light detector 21 ₁ to 21 _3, and outputs a detection signal having a value corresponding to the received light intensity by receiving different periods over to generate light (fluorescence) from each other. In other words, the optical detector 21 ₁ is receiving fluorescence period _{T 11,} the photodetector 21 ₂ receives the fluorescence period _{T 12,} the photodetector 21 ₃ receives fluorescence period _{T 13.} Further, the photodetector 21 ₁ receives the fluorescence period _{T 21,} the photodetector 21 ₂ receives the fluorescence period _{T 22,} the photodetector 21 ₃ receives fluorescence period _{T 23.} Detection signals output from the optical detectors ₂₁ 1 to 21 ₃ is input in common to the input terminal T of the signal converter 31 (inverting input terminal of the amplifier 311).

In the signal processing step, the switch 313 of the signal converter 31 is turned on at the initial _{stage of each of the periods T 11} , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T 23, and the charge accumulation of the capacitance unit 312 is initialized. , The voltage value of the output terminal of the amplifier 311 is also initialized. After initialization, the switch 313 of the signal converter 31 is turned off, and the electric charge is accumulated in the capacitance section 312 based on the detection signal input to the inverting input terminal of the amplifier 311 and is accumulated in the capacitance section 312. A voltage value corresponding to the amount of electric charge is output from the output terminal of the amplifier 311 (signal conversion step). In each of the periods T ₁₁ , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T ₂₃ , the rate of increase of the voltage value output from the signal converter 31 is the detection signal input to the input terminal T of the signal converter 31. It corresponds to the value of, and corresponds to the intensity of the fluorescence received by each photodetector.

At the end of each of the periods T ₁₁ , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T ₂₃ , the voltage value (analog value) output from the signal converter 31 is held by the sample hold circuit 331 of the AD converter 33. , Is output from the sample hold circuit 331 over a certain period of time. Then, it is converted into a digital value by the AD converter 33, and the digital value is output from the AD converter 33 (AD conversion step). The digital value output from the AD converter 33 is transferred to the control unit 40.

Periods T ₁₁ , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T ₂₃ each need only be a time during which an increase in the voltage value output from the signal converter 31 can be observed with a sufficient SN ratio. For example, it is preferably 1 millisecond to 1 minute. For example, if each period is 3 seconds, the measurement time of one cycle in which the measurement is performed once for each of the three samples is 9 seconds.

During the periods T ₁₁ and T ₂₁ in which the light source 11 _{outputs the irradiation light λ 1} , when fluorescence is generated by the first fluorescent substance contained in the sample placed in the sample container 2, the light is detected by receiving the fluorescence. detection signal output from the vessel 21 ₁ is input to the processing unit 30, the value of the detection signal is converted into a digital value. During the periods T ₁₂ and T ₂₂ in which the light source 11 _{outputs the irradiation light λ 2} , when fluorescence is generated by the second fluorescent substance contained in the sample placed in the sample container 2, the light is detected by receiving the fluorescence. detection signal output from the vessel 21 ₂ is input to the processing unit 30, the value of the detection signal is converted into a digital value. Further, during the periods T ₁₃ and T ₂₃ in which the light source 11 _{outputs the irradiation light λ 3} , when fluorescence is generated by the third fluorescent substance contained in the sample placed in the sample container 2, the fluorescence is received. detection signal output from the photodetector 21 ₃ is input to the processing unit 30, the value of the detection signal is converted into a digital value.

The signal processing of the processing unit 30 including the signal converter 31 and the AD converter 33 is performed in synchronization with the output _{of the irradiation lights λ 1 to λ 3 of the light source 11.} The control unit 40 controls _{the output of the irradiation lights λ 1 to λ 3} of the light source 11 and the signal processing of the processing unit 30 in synchronization with each other, so that the digital values output from the processing unit 30 are three types in the sample. It is possible to grasp which of the fluorescent substances in the above represents the intensity of the fluorescence generated by the fluorescent substance. Therefore, the intensity of light generated in each of the three types of fluorescent substances in the sample can be measured. Further, since one processing unit 30 may be provided for each of the _{three photodetectors 21 1 to 213, the apparatus configuration can be simplified.}

FIG. 4 is a timing chart showing another operation example of the optical measuring device 1 of the present embodiment. This operation is also performed based on the synchronization signal output from the control unit 40. Also this figure shows, in order from the top, the irradiation light lambda ₁ to [lambda] ₃ respectively output from the light source 11, the photodetector ₂₁ 1 to 21 ₃ of each output, the output of the output and the AD converter 33 of the signal converter 31 It is shown. Further, although the operation for two cycles is shown in this figure, the same applies to the subsequent cycles.

_{The light source 11 outputs the irradiation light λ 1} during the period T ₁₁ (time t _{10 to t 11} ), and the light source 11 outputs the irradiation light λ ₂ during the period T ₁₂ (time t _{12 to t 13} ) after this period T _11. The light source 11 outputs the _{irradiation light λ 3} in the period T ₁₃ (time t _{14 to t 15} ) after this period T _12. Further, the light source 11 _{outputs the irradiation light λ 1} in the period T ₂₁ (time t _{20 to t 21 ) after this period T 13} , and the period T ₂₂ (time t _{22 to t 23} ) after this period T _21. The light source 11 _{outputs the irradiation light λ 2} and the light source 11 outputs the _{irradiation light λ 3} in the period T ₂₃ (time t _{24 to t 25} ) after this period T _22. The periods T ₁₁ , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ , and T ₂₃ do not overlap each other. In this way, the light source 11 outputs the _{irradiation lights λ 1 to λ 3 over different irradiation periods.}

In this operation example, the waiting period between the _{irradiation period T 11} and the next irradiation period T _{12 (time t 11 to t 12} ) and the waiting period between the irradiation period T ₁₂ and the next irradiation period T ₁₃ (time). t _{13 to t 14} ), waiting period between irradiation period T ₁₃ and next irradiation period T _{21 (time t 15 to t 20} ), waiting period between irradiation period T ₂₁ and next irradiation period T ₂₂ (Times t _{21 to t 22} ), waiting period between irradiation period T ₂₂ and next irradiation period T _{23 (time t 23 to t 24} ) and waiting period between irradiation period T ₂₃ and next irradiation period. At (time t _{25 to t 30} ), the light source 11 does not output irradiation light (excitation light). During these waiting periods, the digital value output from the AD converter 33 is transferred to the control unit 40.

In this operation example, for example, even if delayed fluorescence is generated from the sample after the period T ₁₁ (time t _{10 to t 11 ) in which the light source 11 outputs the irradiation light λ 1} is completed, the generation of delayed fluorescence is awaited. Since it disappears within the period (time t _{11 to t 12} ), the irradiation of the irradiation light λ _{2 from the light source 11 in the next period T 12} (time t _{12 to t 13} ) affects the detection of fluorescence generated in the sample. No.

Next, another configuration example of the signal converter 31 of the processing unit 30 and another configuration example of the processing unit 30 will be described.

FIG. 5 is a diagram showing another configuration example of the signal converter 31 of the processing unit 30. The signal converter 31A of another configuration example shown in this figure is an integrator configuration including an amplifier 311, a capacitance unit 312, and switches 314 to 318. The amplifier 311 has an inverting input terminal, a non-inverting input terminal, and an output terminal. The non-inverting input terminal of the amplifier 311 is set to the ground potential. Switch 314 is provided between the inverting input terminal of the photodetector ₂₁ 1 to 21 _3, respectively the anode and the amplifier 311 of the detection unit 20. The switch 315 is provided between the inverting input terminal and the non-inverting input terminal of the amplifier 311. One end of the capacitance unit 312 is connected to the inverting input terminal of the amplifier 311. The other end of the capacitance unit 312 is connected to the output terminal of the amplifier 311 via the switch 316, and the reference potential Vref is input via the switch 317. The switch 318 is provided between the output terminal of the amplifier 311 and the sample hold circuit 331.

In this signal converter 31A, when the switches 314 and 316 are in the off state and the switches 315 and 317 are in the on state, the voltage between both ends of the capacitance unit 312 is initialized to Vref. When the switches 314 and 316 are on and the switches 315 and 317 are off, electric charges are accumulated in the capacitance section 312 based on the detection signal input to the inverting input terminal of the amplifier 311 and the capacitance is accumulated. A voltage value corresponding to the amount of electric charge stored in the unit 312 is output from the output terminal of the amplifier 311. When the switch 314 turns off, the charge accumulation in the capacitance portion 312 ends. When the switch 318 is in the ON state, the voltage value output from the output terminal of the amplifier 311 is held by the sample hold circuit 331 and is output from the sample hold circuit 331 to the AD converter 33 for a certain period of time.

FIG. 6 is a diagram showing still another configuration example of the signal converter 31 of the processing unit 30. The signal converter 31B of still another configuration example shown in this figure is a configuration of a current-voltage converter including an amplifier 311 and a resistor 319. The amplifier 311 has an inverting input terminal, a non-inverting input terminal, and an output terminal. The non-inverting input terminal of the amplifier 311 is set to the ground potential. The resistor 319 is provided between the inverting input terminal and the output terminal of the amplifier 311. In this signal converter 31B, a voltage value corresponding to the current value of the detection signal input to the inverting input terminal of the amplifier 311 is output from the output terminal.

FIG. 7 is a timing chart showing an operation example of the optical measuring device 1 when the signal converter 31B is used. This operation is also performed based on the synchronization signal output from the control unit 40. Also this figure shows, in order from the top, the irradiation light lambda ₁ to [lambda] ₃ respectively output from the light source 11, each output optical detectors ₂₁ 1 to 21 _3, the output of the output and the sample and hold circuit 331 of the signal converter 31B is It is shown. Further, although the operation for two cycles is shown in this figure, the same applies to the subsequent cycles.

Each of the output illumination light lambda ₁ to [lambda] ₃ and the photodetector 21 ₁ to 21 ₃ of each output from the light source 11 in the operation example shown in FIG. 7 is the same as that of the operation example shown in FIG. 3 .. In the operation example shown in FIG. 7, the voltage value output from the output terminal of the signal converter 31B is the voltage value of the signal converter 31B in each of _{the periods T 11} , T ₁₂ , T ₁₃ , T ₂₁ , T ₂₂ and T _23. This corresponds to the current value of the detection signal input to the inverting input terminal of the amplifier 311. If the input current value of the signal converter 31B is constant in each period, the output voltage value of the signal converter 31B is also constant. In this case, the voltage value (analog value) output from the signal converter 31B at an arbitrary timing within each period is held by the sample hold circuit 331, and is output from the sample hold circuit 331 to the AD converter 33 over a certain period of time. Will be done. After that, the AD converter 33 converts the voltage value output from the sample hold circuit 331 into a digital value, and the digital value is output from the AD converter 33. The digital value output from the AD converter 33 is transferred to the control unit 40.

FIG. 8 is a diagram showing another configuration example of the processing unit 30. The processing unit 30A of another configuration example shown in this figure includes a signal converter 31, a signal converter 32, an AD converter 33, a switch 34, and a switch 35.

The signal converter 31 has an integrator configuration including an amplifier 311, a capacitance unit 312 and a switch 313, similar to that shown in FIG. The signal converter 32 also has an integrator configuration including an amplifier 321 and a capacitance unit 322 and a switch 323.

The switch 34 selects either one of the input terminal T1 of the signal converter 31 (the inverting input terminal of the amplifier 311) and the input terminal T2 of the signal converter 32 (the inverting input terminal of the amplifier 321), and selects one of them. electrically connecting the input terminal and the optical detector 21 ₁ to 21 ₃ of each anode of the detecting section 20. The switch 35 selects either one of the output terminal of the signal converter 31 (output terminal of the amplifier 311) and the output terminal of the signal converter 32 (output terminal of the amplifier 321), and sets the selected output terminal. It is electrically connected to the sample hold circuit 331. The connection state of the switch 34 and the switch 35 is controlled by the control unit 40.

FIG. 9 is a timing chart showing an operation example of the optical measuring device 1 when the processing unit 30A is used. This operation is also performed based on the synchronization signal output from the control unit 40. In this figure, in order from the top, _{the outputs of the irradiation lights λ 1 to λ 3} from the light source 11, the connection state of the switch 34, the connection state of the switch 35, the operation of the signal converter 31, the operation of the signal converter 32, and the operation of the signal converter 32. The operation of the AD converter 33 is shown.

During the period T ₁₁ (time t _{10 to t 12} ), the light source 11 _{outputs the irradiation light λ 1} , the photodetector 21 ₁ receives the fluorescence, and the input terminal T 1 of the signal converter 31 detects each light by the switch 34. It is connected to the vessel 21 _n. At the beginning of this period T ₁₁ , the switch 313 of the signal converter 31 is turned on, the charge accumulation of the capacitance unit 312 is initialized, and the voltage value of the output terminal of the amplifier 311 is also initialized. After initialization, the switch 313 of the signal converter 31 is turned off, and the electric charge is accumulated in the capacitance section 312 based on the detection signal input to the inverting input terminal of the amplifier 311 and is accumulated in the capacitance section 312. A voltage value corresponding to the amount of electric charge is output from the output terminal of the amplifier 311.

During the period T ₁₂ (time t _{12 to t 14} ) following this period T ₁₁ , the input terminal T1 of the signal converter 31 is _{disconnected from each photodetector 21 n} by the switch 34, and the output voltage value of the signal converter 31 is set. Is confirmed. Further, during this period T ₁₂ , the output terminal of the signal converter 31 is connected to the sample hold circuit 331 by the switch 35, and the output voltage value of the signal converter 31 is held by the sample hold circuit 331. During this period T ₁₂ , the AD converter 33 performs the AD conversion process and outputs the digital value to the control unit 40. Digital value output at this time are those corresponding to the detection signal value of the photodetector 21 _1.

Further, in a period _{T 12,} the light source 11 outputs illumination light lambda _2, the optical detector 21 ₂ is receiving fluorescence, input terminal T2 of the signal converter 32 is connected to the respective optical detectors 21 _n by the switch 34 NS. At the beginning of this period T ₁₂ , the switch 323 of the signal converter 32 is turned on, the charge accumulation of the capacitance unit 322 is initialized, and the voltage value of the output terminal of the amplifier 321 is also initialized. After initialization, the switch 323 of the signal converter 32 is turned off, and the electric charge is accumulated in the capacitance section 322 based on the detection signal input to the inverting input terminal of the amplifier 321 and is accumulated in the capacitance section 322. A voltage value corresponding to the amount of electric charge is output from the output terminal of the amplifier 321.

During the period T ₁₃ (time t _{14 to t 20} ) following this period T ₁₂ , the input terminal T2 of the signal converter 32 is _{disconnected from each photodetector 21 n} by the switch 34, and the output voltage value of the signal converter 32 is changed. Is confirmed. Further, during this period T ₁₃ , the output terminal of the signal converter 32 is connected to the sample hold circuit 331 by the switch 35, and the output voltage value of the signal converter 32 is held by the sample hold circuit 331. During this period T ₁₃ , AD conversion processing is performed by the AD converter 33, and the digital value is output to the control unit 40. Digital value output at this time are those corresponding to the detection signal value of the light detector 21 _2.

Further, in a period _{T 13,} the light source 11 outputs illumination light lambda _3, the optical detector 21 ₃ receives the fluorescence, an input terminal T1 of the signal converter 31 is connected to the respective optical detectors 21 _n by the switch 34 NS. At the beginning of this period T ₁₃ , the switch 313 of the signal converter 31 is turned on, the charge accumulation of the capacitance unit 312 is initialized, and the voltage value of the output terminal of the amplifier 311 is also initialized. After initialization, the switch 313 of the signal converter 31 is turned off, and the electric charge is accumulated in the capacitance section 312 based on the detection signal input to the inverting input terminal of the amplifier 311 and is accumulated in the capacitance section 312. A voltage value corresponding to the amount of electric charge is output from the output terminal of the amplifier 311.

During the period T ₂₁ (time t _{20 to t 22} ) following this period T ₁₃ , the input terminal T1 of the signal converter 31 is _{disconnected from each photodetector 21 n} by the switch 34, and the output voltage value of the signal converter 31 is set. Is confirmed. Further, during this period T ₂₁ , the output terminal of the signal converter 31 is connected to the sample hold circuit 331 by the switch 35, and the output voltage value of the signal converter 31 is held by the sample hold circuit 331. During this period T ₂₁ , AD conversion processing is performed by the AD converter 33, and the digital value is output to the control unit 40. Digital value output at this time are those corresponding to the detection signal value of the photodetector 21 _3.

Subsequent operations are the same. In this configuration example, the two signal converters 31 and 32 alternately input detection signals to accumulate charges in synchronization with the irradiation periods _{of the irradiation lights λ 1 to λ 3 of the light source 11.} During the period when one signal converter inputs the detection signal and stores the charge, the AD converter 33 digitally sets the output voltage value of the other signal converter that stores the charge in the previous period. And output the digital value. Therefore, even while the AD converter 33 converts the output voltage value of one signal converter into a digital signal, the charge can be accumulated by the other signal converter.

The arrangement and configuration of the light source, the sample container, the photodetector, and other optical components in the measurement optical system are not limited to the above-described embodiment. For example, the irradiation light may be guided by an optical fiber from the light source to the sample container, or the generated light may be guided by an optical fiber from the sample container to the optical detector. Further, as shown in FIG. 10, in the light source 11 and the photodetector 21 measuring optical system, including _n, between the sample container 2 and the photodetector 21 _n, in addition to the fluorescence transmitting filter 22 _n lens 23 _n , 24 _n may be provided. The fluorescence transmission filter 22 _n is preferably provided between the lens 23 _n and the lens 24 _n. In this configuration example, by the lens 23 _n, 24 _n for selectively guided to the photodetector 21 _n fluorescence generated by the sample is provided, the generated light efficiently photodetector 21 _n while capable of directing the light receiving surface, it is possible to suppress the excitation light or the scattered light is incident on the light receiving surface of the photodetector 21 _n, the intensity of the generated light can be detected with high SN ratio ..

However, the measurement optical system including the lens as shown in FIG. 10 has a complicated configuration and a large number of man-hours for assembly, which causes an increase in the price of the optical measurement device. Therefore, it is preferable to configure the measurement optical system as shown below.

FIG. 11 is a vertical cross-sectional view showing the configuration of a preferred example of the measurement optical system. The dark box 80 is also shown in this figure. FIG. 12 is a vertical cross-sectional view showing the configuration of the metal block 50 in the measurement optical system shown in FIG. FIG. 13 is a vertical cross-sectional view showing the configuration of the optical element holding portion 60 in the measurement optical system shown in FIG.

Metal block 50 and the optical element holder 60, the sample container 2, the light source 11, the excitation light transmission filter 12, and holds the optical detector ₂₁ 1 to 21 ₃ and the fluorescence transmitting filter ₂₂ 1-22 _3, these arrangements It stipulates. Dark box 80 is arranged sample container 2, the light source 11, the excitation light transmitting filter 12, an optical detector ₂₁ 1 to 21 _3, the fluorescence transmitting filter ₂₂ 1-22 _3, the metal block 50 and the optical element holder 60 therein. With the lid 81 of the dark box 80 open, the sample container 2 can be inserted and removed from the first through hole extending in the vertical direction of the metal block 50. When the lid 81 is closed, stray light from the outside does not enter the inside of the dark box 80.

The metal block 50 shown in FIG. 12 is made of a metal having high thermal conductivity, and is preferably composed of at least one of aluminum and an aluminum alloy. Further, the metal block 50 may be made of copper, a copper alloy, stainless steel, or the like. The metal block 50 has a prismatic shape as an approximate shape. The metal block 50 has a first surface 51 which is an upper surface of the prism, a second surface 52 which is a lower surface facing the first surface 51, and a third surface 53 which is a side surface different from these. If the approximate shape of the metal block 50 is a prismatic shape, there are at least three third surfaces 53, which are side surfaces. For example, if the approximate shape of the metal block 50 is a quadrangular prism shape, there are four third surfaces 53 that are side surfaces.

The metal block 50 has a first through hole 54 provided between the first surface 51 and the second surface 52. The first through hole 54 is divided into an upper hole 55 on the first surface 51 side, a tapered portion 56, and a lower hole 57 on the second surface 52 side. The diameter of the upper hole 55 is constant along the axial direction. The diameter of the lower hole 57 may also be constant along the axial direction. The diameter of the upper hole 55 is smaller than the diameter of the lower hole 57. The upper hole 55 has a diameter into which the sample container 2 can be inserted from the side of the first surface 51. The diameter of the tapered portion 56 between the upper hole 55 and the lower hole 57 changes monotonically along the axial direction. The metal block 50 has a second through-hole ₅₈ 1-58 ₃ provided between either the third surface 53 of the lower hole 57 of the first through hole 54. The diameter of each second through hole 58 _n may be constant. The metal block 50 having a shape can be manufactured by forming a first through hole 54 and second through-holes 58 ₁ to 58 ₃ by the turning to the metal隗.

The sample container 2 is inserted into the first through hole 54 from the side of the first surface 51 of the metal block 50. The upper part of the sample container 2 to be inserted has a substantially cylindrical shape. The lower portion of the sample container 2 may have a shape in which the diameter becomes smaller toward the tip. The inserted sample container 2 is positioned so that the sample 3 in the lower portion thereof faces each of the second through holes 58 _n. At the same time, the substantially cylindrical portion of the upper part of the sample container 2 is in thermal contact with the inner wall surface of the upper hole 55 of the first through hole 54.

There is a microtube as a sample container having the above-mentioned shape. For example, the capacity of a microtube is 250 μL, and the sample placed in the microtube is several μL. The microtube is transparent or milky white. The milky white microtube not only mirror-reflects the excitation light on the surface, but also scatters the excitation light. Since the excitation light scattered by the microtube has a low intensity, it can be sufficiently blocked by the _{fluorescence transmission filter 22 n.} Excitation light specularly reflected by the micro tube, so strength than scattered excitation light is large, a part can not be blocked sufficiently by fluorescence transmitting filter 22 _n when incident on the fluorescent transmission filter 22 _n It may reach the light receiving surface of the light detector 21 _n , or fluorescence may be generated in the fluorescence _{transmission filter 22 n and} the fluorescence may reach the light receiving surface of the light _{detector 21 n.} Therefore, structures such as excitation light specularly reflected by the microtube is not incident on the fluorescence transmitting filter 22 _n are preferred.

The optical element holding portion 60 shown in FIG. 13 is made of a resin having a lower thermal conductivity than the metal block 50 such as DURACON (registered trademark) manufactured by Polyplastics. Optical element holding portion 60 includes a light source 11, the excitation light transmission filter 12, and holds the optical detector ₂₁ 1 to 21 ₃ and the fluorescence transmitting filter ₂₂ 1-22 _3, defining these arrangements. By combining the optical element holding portion 60 and the metal block 50 with each other, the measurement optical system from the light source 11 to the photodetector 21 _{n via the excitation light transmission filter 12, the sample container 2 and the fluorescence transmission filter 22 n can be formed.} It is composed.

The optical element holding portion 60 has a shape that can be combined with the metal block 50. The optical element holding portion 60 has a first through hole 61 that connects with the first through hole 54 of the metal block 50 when combined with the metal block 50. The first through hole 61 of the optical element holding portion 60 is divided into an upper hole 62 near the metal block 50 and a lower hole 63 on the opposite side of the metal block 50. An excitation light transmission filter 12 is fitted in the upper hole 62 to fix the position. A light source 11 is arranged in the opening portion of the lower hole 63.

_{Further, the optical element holding portion 60 has a second through hole 64 n} that is connected to the second through hole _{58 n} of the metal block 50 when combined with the metal block 50. _{The second through hole 64 n} of the optical element holding portion 60 is divided into an inner hole 65 _n close to the metal block 50, a relay hole 66 _{n, and an outer hole 67 n} on the opposite side of the metal block 50. A fluorescence transmission filter 22 _n is fitted into the inner hole 65 _{n to fix the position.} A photodetector 21 _n is fitted into the outer hole 67 _{n to fix the position.}

Due to the configuration of the metal block 50 and the optical element holding portion 60, the light source 11 provided on the side of the second surface 52 of the metal block 50 irradiates the sample 3 in the sample container 2 with the irradiation light λ _1. The irradiation light λ _n selected from ~ λ ₃ is output. The excitation light transmission filter 12 provided on the optical path between the light source 11 and the sample container 2 selectively transmits the excitation light of a specific wavelength among the light output from the light source 11 and causes the excitation light to enter the sample container 2. .. _{The fluorescence transmission filter 22 n} provided on the optical path between the sample container 2 and the photodetector 21 _n selectively blocks the excitation light and selectively transmits the fluorescence to receive the light received by the photodetector _{21 n.} Make it incident on the surface. _{Further, the photodetector 21 n} provided on the side of the third surface 53 of the metal block 50 has a light receiving surface that receives fluorescence generated in response to the irradiation of the irradiation light in the sample 3 in the sample container 2. , Detects the intensity of fluorescence that has reached the light receiving surface through the second through hole 58 _n of the metal block 50 and the fluorescence transmission filter 22 _{n, and outputs a detection signal.}

FIG. 14 is a cross-sectional view of the measurement optical system shown in FIG. This figure shows a cross section at a position where the second through hole 58 _{n of the metal block 50 and the second through hole 64 n} of the optical element holding portion 60 are provided. Further, in this figure, the metal block 50 has a substantially quadrangular prism shape. The first side of the four sides of the metal block 50, the second through-hole 58 1 for detecting fluorescence generated by the irradiation of the irradiation light lambda ₁ to the sample _3, the photodetector 21 ₁ and fluorescence transmitting filter 22 ₁ is provided. To a second aspect, the second through-hole 58 2 for detecting fluorescence generated by the irradiation of the irradiation light lambda ₂ to the sample _3, the light detector 21 ₂ and the fluorescent transmission filter 22 ₂ is provided. To a third aspect, the second through-hole 58 3 for detecting fluorescence generated by the irradiation of the irradiation light lambda ₃ to the sample _3, the photodetector 21 ₃ and fluorescent transmitting filter 22 ₃ are provided.

Further, a temperature adjusting unit 70 is provided in contact with the fourth side surface of the metal block 50. The temperature adjusting unit 70 adjusts the temperature of the metal block 50 and also adjusts the temperature of the sample 3 in the sample container 2 held by the metal block 50. The temperature adjusting unit 70 includes, for example, a heating unit including a heater or a Pertier element, and a temperature measuring unit including, for example, a thermocouple, a thermistor, or a temperature measuring IC, and the temperature of the metal block 50 measured by the temperature measuring unit is desired. Heat by the heating part so that it is within the range. The operation of the temperature adjusting unit 70 may be controlled by the control unit 40.

Next, the operation in the case of the configuration shown in FIGS. 11 to 14 will be described. After the temperature of the metal block 50 is set to a desired range by the temperature adjusting unit 70, the sample container 2 in which the sample 3 is placed is a hole on the upper side of the first through hole 54 from the side of the first surface 51 of the metal block 50. It is inserted in 55. After this insertion, the lid 81 is closed to prevent stray light from entering the inside of the dark box 80 from the outside. This insertion sample container 2, together with the sample 3 is positioned such that at a position facing the second through-hole 58 _{1-58 3,} in thermal contact with the inner wall surface of the upper hole 55. The temperature of the sample 3 in the sample container 2 that is in thermal contact with the inner wall surface of the upper hole 55 gradually reaches a desired range.

The irradiation light λ _{n selected from the irradiation light λ 1 to λ 3} and output from the light source 11 passes through the lower hole 63 of the first through hole 61 of the optical element holding portion 60, and passes through the first through hole 61. The excitation light transmission filter 12 fitted in the upper hole 62 of the above is transmitted. The excitation light that has passed through the excitation light transmission filter 12 passes through the lower hole 57 of the first through hole 54 of the metal block 50 and irradiates the sample 3 in the sample container 2. A part of the fluorescence generated in the sample 3 in response to the irradiation of the sample 3 with the excitation light _{passes through the second through hole 58 n} of the metal block 50, and the inner hole _{of the second through hole 64 n} of the optical element holding portion 60. It transmits the fluorescence transmission filter 22 _{n fitted in 65 n} . The fluorescence transmitted through the fluorescence transmission filter 22 _n passes through the relay hole 66 _{n of the second through hole 64 n} of the optical element holding portion 60, and is a photodetector fitted in the outer hole 67 _n of the second through hole 64 _n. The light is received by _{21 n.}

_{Irradiation lights λ 1 to λ 3} are output from the light source 11 over different irradiation periods. The process of the irradiation light lambda ₁ to [lambda] ₃ respectively output and the photodetector 21 ₁ to 21 ₃ detection signals output from the light source 11 is as already described.

The lower hole 63 of the first through hole 61 of the optical element holding portion 60 and the lower hole 57 of the first through hole 54 of the metal block 50 through which the excitation light passes from the light source 11 to the sample container 2, respectively. The size of the sample container 2 may be about the same as or slightly larger than the diameter of the sample container 2. _{The size of the second through hole 58 n} of the metal block 50 through which fluorescence passes from the sample 3 to the photodetector 21 _n and the relay hole 66 _n of the second through hole 64 _n of the optical element holding portion 60 are light. it may be sized approximately the same light-receiving surface of the detector 21 _n (for example, about 5 mm). The length of the second through hole 58 _n of the metal block 50 may be several mm.

As described above, in the configuration shown in FIGS. 11 to 14, by inserting the sample container 2 into the upper hole 55 of the first through hole 54 of the metal block 50, the sample 3 in the sample container 2 is second. The sample container 2 can be positioned so as to face the through hole 58 _n. As a result, a measurement optical system is configured from the light source 11 through the excitation light transmission filter 12, the sample container 2 and the fluorescence transmission filter 22 _n to the photodetector 21 _n. Further, the fluorescent optical system from the sample container 2 to the photodetector 21 _n is not provided with a lens. The fluorescence generated in the sample 3 in the sample container 2 _{passes through the second through hole 58 n} of the metal block 50, passes through the fluorescence transmission filter 22 _n, and is received by the photodetector _{21 n.} Therefore, since the configuration is simple and the assembly man-hours are small, it is easy and the price can be reduced.

In this configuration, the sample container 2 is inserted into the upper hole 55 of the first through hole 54 of the metal block 50 to position the sample container 2 and at the same time, the sample container 2 is brought into thermal contact with the inner wall surface of the upper hole 55. .. Therefore, the temperature adjusting unit 70 can detect the intensity of fluorescence generated in the sample 3 under the condition that the temperature of the sample 3 in the sample container 2 is set within a desired range via the metal block 50. , The change in fluorescence intensity over time can be observed.

By using a colored glass filter as the excitation light transmission filter 12 or by using a colored glass filter as the fluorescence transmission filter 22 _n , the light measuring device 1 can be further reduced in price.

The colored glass filter as the fluorescence transmission filter 22 _n may generate fluorescence when it absorbs the excitation light, and when the fluorescence is _{received by the photodetector 21 n} , the SN ratio becomes worse. In response to this problem, the second through hole 58 _n of the metal block 50 functions as a collimating function that defines the direction of light incident on the photodetector 21 _n , and the fluorescence generated in the sample 3 receives the light received by the photodetector _{21 n.} while allowed to enter the plane, toward the light receiving surface of the photodetector 21 _n after the excitation light transmitted through the excitation light transmission filter 12 is outputted from the light source 11 is specularly reflected by direct or sample container 2 It is possible to suppress the propagation of light. Therefore, even when a fluorescent _{colored glass filter is used as the fluorescence transmission filter 22 n} , deterioration of the SN ratio is suppressed. The second through hole 58 _n of the metal block 50 is directed to the light receiving surface of the photodetector _{21 n} after the excitation light output from the light source 11 and transmitted through the excitation light transmission filter 12 is directly reflected or specularly reflected by the sample container 2. It is preferably a position, diameter, length or orientation that limits propagation towards.

A configuration for excitation light transmitted through the excitation light transmission filter 12 is prevented from being propagated toward the light receiving surface of the photodetector 21 _n, the metal block 50, the excitation light on the inner wall surface of the first through hole 54 also has a to suppress the reflected or scattered light trap that limits the propagation of the excitation light toward the light receiving surface of the photodetector 21 _n in the first through-hole 54 preferably. As shown in FIG. 11, the tapered portion 56 of the first through hole 54 of the metal block 50 gradually decreases in diameter from the lower hole 57 to the upper hole 55, and the distance from the sample container 2 gradually decreases. The smaller size constitutes the optical trap section. This also even in the case of using a colored glass filter having fluorescence as fluorescence transmitting filter 22 _n, the deterioration of SN ratio can be suppressed.

As in the configuration of the first modification of the measurement optical system shown in FIG. 15, the metal block 50 has a first through hole of _{a shielding portion 59 n} that shields the propagation of excitation light toward the light receiving surface of the photodetector _{21 n.} It is also preferable to have it in 54. The shielding portion 59 _n is provided in the first through hole 54 and below _{the second through hole 58 n.} This shielding portion 59 _n is exert a collimating function of defining a light incident direction of the photodetector 21 _n, while the fluorescence generated in the sample 3 is allowed to enter the light receiving surface of the photodetector 21 _n, it can be suppressed to propagate toward the light receiving surface of the photodetector 21 _n after the excitation light transmitted through the excitation light transmission filter 12 is specularly reflected by direct or sample container 2. This also even in the case of using a colored glass filter having fluorescence as fluorescence transmitting filter 22 _n, the deterioration of SN ratio can be suppressed.

As in the configuration of the second modification of the measurement optical system shown in FIG. 16, the photodetector 21 _n has a perpendicular direction of the light receiving surface with respect to the optical axis direction of the excitation light from the light source 11 to the sample container 2. It is also preferable that the light is arranged so as to have a sharp angle. By thus receiving surface of the photodetector 21 _n is inclined, the light receiving surface of the photodetector 21 _n after the excitation light transmitted through the excitation light transmission filter 12 is specularly reflected by direct or sample container 2 The incident angle (angle of the light receiving surface with respect to the perpendicular line) at the time of incident on the light is increased, and the light receiving sensitivity to the excitation light is lowered. This also even in the case of using a colored glass filter having fluorescence as fluorescence transmitting filter 22 _n, the deterioration of SN ratio can be suppressed.

Next, a configuration example of the light source 11 will be described.

FIG. 17 is a perspective view showing a first configuration example of the light source 11. Light source 11A of the first structural example has a structure in which the light-emitting element ₉₁ 1-91 ₃ 3 on the substrate 90 is flip-chip mounted. Each light emitting element 91 _n can output the irradiation light λ _n. Each light emitting element 91 _n is, for example, an LED (light emitting diode) or an LD (laser diode). One electrode terminal of each light emitting element 91 _n is electrically connected to _{the wiring W n 1 on the substrate 90.} The other electrode terminal of each light emitting element 91 _n is electrically connected to _{the wiring W n 2 on the substrate 90.} On the substrate 90, _{the distance between the three light emitting elements 91 1 to} 913 is preferably short, and the distance can be, for example, 1 mm or less.

Light source drive circuit 13 which is controlled by the control unit 40, wiring sets _{W 11, W 12,} wiring sets _{W 21, W 22,} and selects one of the wiring sets from among the wiring sets _{W 31, W 32} by supplying a driving current between wire _{W n1, W n2} that the selection, it is possible to output the illumination light lambda _n from the wire _{W n1, W n2} to the connected light emitting elements 91 _n.

Provided lens so as to cover the light emitting element 91 _{1-91 3} on the light output side of the light source 11A, it is preferable to suppress the divergence of the illumination light lambda _n by the lens. This lens has been molded by a resin, may be integral with the substrate 90 and the light emitting element 91 _{1-91 3} may be such as are separate bodies.

FIG. 18 is a cross-sectional view showing a second configuration example of the light source 11. Light source 11B of the second configuration example has a configuration in which the light-emitting element ₉₂ 1-92 ₃ and the dichroic mirror ₉₃ 1, 93 ₂ is held by the light emitting element holding member 94. Each light emitting element 92 _n can output the irradiation light λ _n. Each light emitting element 92 _n is, for example, an LED (light emitting diode) or an LD (laser diode).

Emitting element holding member 94 has a through-hole 94 _1. One end of the through hole 94 _first light emitting element 92 ₁ is fitted, the irradiation light is outputted from the other end of the through-hole 94 _1. Further, the light emitting element holding member 94 has through holes 94 ₂ and 94 _{3 extending from the side surface to the through holes 94 1} . Is the light emitting element 92 ₂ is fitted into the through holes 94 _2, and the through-hole 94 ₂ and the through-hole 94 ₁ is the dichroic mirror 93 ₁ is located where. Light emitting element 92 ₃ in the through hole 94 ₃ is fitted, the through-holes 94 ₃ and the through-hole 94 ₁ is the dichroic mirror 93 ₂ are located where.

The dichroic mirror 93 _1, the light emitting element 92 is transmitted through the irradiation light lambda ₁ output from the _1, reflects the irradiated light lambda ₂ outputted from the light emitting element 92 _2. The dichroic mirror 93 _2, is transmitted through the dichroic mirror 93 ₁ irradiation light lambda ₁ arriving from or irradiation light lambda _2, and reflects the irradiated light lambda ₃ outputted from the light emitting element 92 _3. Then, each of the irradiation light lambda _n is outputted from the through hole 94 ₁ to the outside.

FIG. 19 is a cross-sectional view showing a third configuration example of the light source 11. FIG. 20 is a perspective view showing the filter wheel 96 used in the third configuration example of the light source 11. The light source 11C of the third configuration example includes a light emitting element 95, a filter wheel 96, and a stepping motor 98. The light emitting element 95 _{outputs a wide band white light λ W} , and is, for example, an SLD (superluminescent diode). The filter wheel 96 has a substantially disk shape. The filter wheel 96 is coupled to the stepping motor 98 by a rotating shaft 99, and is rotatable around the center by the stepping motor 98.

Filter 97 _{1-97 8} is provided from the center of rotation of the filter wheel 96 on the circumference of a predetermined distance. Filter ₉₇ 1-97 ₈ respectively, have different transmission characteristics from each other. Filter 97 _{1-97 8} respectively, enter the white light lambda _W output from the light emitting element 95 can output the illumination light lambda _n having a spectrum corresponding to the transmission characteristic. In the light source 11C, the control unit 40 by the rotation of the stepping motor 98 is controlled, the white light lambda _W is inputted to one of the filter of the filters 97 ₁ to 97 _8, the irradiation light from the filter λ _n is output.

The present invention is not limited to the above embodiment, and various modifications are possible. The number of each of the irradiation light and the photodetector is set to 3 in the above embodiment, but may be 2 or 4 or more. The generated light generated in the sample irradiated with light is considered to be fluorescent in the above embodiment, but may be scattered light or light generated by a nonlinear optical phenomenon. A filter that selectively transmits light of a specific wavelength among the light output from the light source and a filter that selectively transmits light of a specific wavelength among the light generated by the sample may be provided as necessary.

Various other configuration examples are possible for the light source 11. For example, the light source 11 may be a tunable light source in which the output light wavelength is variable and adjustable. For example, as a tunable light source, a semiconductor light emitting element capable of emitting light in a wide band may be arranged on an optical path in an external resonator, and the external resonator may have an oscillation wavelength adjustment function.

1 ... Optical measuring device, 2 ... Sample container, 3 ... Sample, 10 ... Irradiation unit, 11, 11A to 11C ... Light source, 12 ... Excitation light transmission filter, 13 ... Light source drive circuit, 20 ... Detection unit, 21 _{1 to} 21 ₃ ... Optical detector, 22 _{1 to} 22 ₃ ... Fluorescence transmission filter, 30, 30A ... Processing unit, 31, 31A, 31B, 32 ... Signal converter, 33 ... AD converter, 40 ... Control unit, 41 ... Input unit , 42 ... Display unit, 43 ... Storage unit, 50 ... Metal block, 51 ... 1st surface, 52 ... 2nd surface, 53 ... 3rd surface, 54 ... 1st through hole, 55 ... Upper hole, 56 ... Tapered part , 57 ... Lower hole, 58 _{1 to} 58 ₃ ... Second through hole, 59 _{1 to} 59 ₃ ... Shielding part, 60 ... Optical element holding part, 61 ... First through hole, 62 ... Upper hole, 63 ... Lower side Holes, 64 ₁ to 64 ₃ ... 2nd through hole, 65 ... Inner hole, 66 ... Relay hole, 67 ... Outer hole, 70 ... Temperature control unit, 80 ... Dark box, 81 ... Lid, 90 ... Substrate, 91 _{1 to 91 3} ... light emitting element, ₉₂ 1-92 3 _... light emitting element, ₉₃ 1, 93 2 _... dichroic mirror, 94 ... light emitting element holding member, 95 ... light emitting element, 96 ... filter wheel, ₉₇ 1-97 8 _... filter, 98 ... Stepping motor.

Claims (13)

A device that measures the intensity of light generated in a sample in response to light irradiation on the sample.
It includes a light source that outputs irradiation light of any one peak wavelength selected from a plurality of peak wavelengths, and irradiates the sample with irradiation light of each peak wavelength output from the light source over different irradiation periods. Irradiation part and
A plurality of light detectors provided in a one-to-one correspondence with the plurality of peak wavelengths are included, and the generated light generated in the sample according to the irradiation of the irradiation light of the corresponding peak wavelength by each light detector. Among them, a detection unit that receives generated light emitted in a direction different from the irradiation direction of the irradiation light, detects the intensity of the received generated light, and outputs a detection signal.
It has an input terminal in which detection signals output from each of the plurality of optical detectors are commonly input as analog values, and is input to the input terminal during an irradiation period in which the light source outputs irradiation light of each peak wavelength. A processing unit that inputs an analog value to be input and outputs a digital value corresponding to the analog value as a value indicating the intensity of the generated light generated in the sample in response to the irradiation of the irradiation light of the peak wavelength.
Bei to give a,
The processing unit has a signal converter that outputs a voltage value corresponding to the value of the detection signal input to the input terminal, and a signal converter that converts the voltage value output from the signal converter into a digital value and converts the digital value into a digital value. Including the AD converter to output
The signal converter has a capacitance unit that accumulates electric charges based on a detection signal input to the input terminal for each irradiation period, and outputs a voltage value according to the amount of accumulated electric charges for each irradiation period. Including the integrator to
Optical measuring device. The irradiation unit outputs irradiation light of each peak wavelength from the light source over irradiation periods different from each other based on a synchronization signal indicating an irradiation period of irradiation light of each peak wavelength.
The processing unit performs conversion processing from an analog value to a digital value based on the synchronization signal.
The optical measuring device according to claim 1. A control unit that outputs the synchronization signal to the irradiation unit and the processing unit is further provided.
The optical measuring device according to claim 2. The signal converter includes the integrator which is the first integrator and the integrator which is the second integrator.
The first integrator and the second integrator alternately input the detection signals in synchronization with the irradiation period of each of the irradiation lights having the plurality of peak wavelengths.
The optical measuring device according to any one of claims 1 to 3. The irradiation unit includes the light source having a plurality of light emitting elements having different peak wavelengths of output light, and outputs irradiation light from any one of the plurality of light emitting elements selected from the plurality of light emitting elements to irradiate the light source. Irradiate the sample with light,
The optical measuring device according to any one of claims 1 to 4. The irradiation unit includes the light source having a variable output light wavelength, outputs irradiation light of any one peak wavelength selected from a plurality of peak wavelengths from the light source, and outputs the irradiation light to the sample. Irradiate,
The optical measuring device according to any one of claims 1 to 4. A method of measuring the intensity of light generated in a sample in response to light irradiation of the sample.
In an irradiation unit including a light source that outputs irradiation light of any one peak wavelength selected from a plurality of peak wavelengths, the sample is provided with irradiation light of each peak wavelength output from the light source over different irradiation periods. Light irradiation step to irradiate
In the detection unit including a plurality of photodetectors provided in a one-to-one correspondence with the plurality of peak wavelengths, each photodetector generates light in the sample according to the irradiation of the irradiation light of the corresponding peak wavelength. A photodetection step of receiving generated light emitted in a direction different from the irradiation direction of the irradiation light, detecting the intensity of the received generated light, and outputting a detection signal.
In a processing unit having an input terminal in which detection signals output from each of the plurality of optical detectors are commonly input as analog values, the input terminal is used during an irradiation period in which the light source outputs irradiation light of each peak wavelength. A signal processing step in which an analog value input to is input and a digital value corresponding to the analog value is output as a value representing the intensity of the generated light generated in the sample in response to the irradiation of the irradiation light of the peak wavelength. ,
Only including,
The signal processing step includes a signal conversion step that outputs a voltage value corresponding to the value of the detection signal input to the input terminal, and an AD that converts the output voltage value into a digital value and outputs the digital value. Including conversion steps,
In the signal conversion step, the irradiation value corresponding to the amount of accumulated electric charge is applied by using an integrator having a capacitance part that accumulates electric charge based on the detection signal input to the input terminal for each irradiation period. Output every period,
Optical measurement method. In the light irradiation step, the irradiation light of each peak wavelength is output from the light source over different irradiation periods based on the synchronization signal indicating the irradiation period of the irradiation light of each peak wavelength.
In the signal processing step, conversion processing from an analog value to a digital value is performed based on the synchronization signal.
The optical measurement method according to claim 7. A synchronization step of outputting the synchronization signal to the irradiation unit and the processing unit is further included.
The optical measurement method according to claim 8. In the signal conversion step, the two integrators are used to alternately accumulate charges in synchronization with the irradiation period of each of the irradiation lights having the plurality of peak wavelengths.
The optical measurement method according to any one of claims 7 to 9. In the light irradiation step, the light source having a plurality of light emitting elements having different peak wavelengths of the output light is used, and the irradiation light is output from any one of the plurality of light emitting elements selected from the plurality of light emitting elements, and the irradiation light is output. Irradiating the sample with irradiation light,
The optical measurement method according to any one of claims 7 to 10. In the light irradiation step, the light source having a variable output light wavelength is used, irradiation light of any one peak wavelength selected from a plurality of peak wavelengths is output from the light source, and the irradiation light is used as the sample. Irradiate to
The optical measurement method according to any one of claims 7 to 10. The generated light is light generated by fluorescence, scattered light, or a nonlinear optical phenomenon.
The optical measurement method according to any one of claims 7 to 12.

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