JPH06100533B2 - Chromatoskiana - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial field of application) The present invention is for optically quantifying a sample fraction separated on a sample plate such as a thin layer plate of thin layer chromatography (TLC) or an electrophoresis gel. It relates to a chromatographic scanner used in.

(Prior Art) In a conventional chromatographic scanner, a sample plate to be measured is fixed with a light beam to be irradiated, and a stage on which the sample plate is mounted is moved in X and Y directions.
The absorption of the sample spot is measured so that the irradiation light beam scans the sample plate in a zigzag pattern.

(Problems to be Solved by the Invention) In the method of moving the stage, there is a limit in increasing the scanning speed because the inertia of the stage is large.

The present invention is a method of scanning a light beam irradiated on a sample plate, the light beam detected by scanning the light beam moves on the detector, and variations in sensitivity due to the position on the light receiving surface of the detector, Since so-called locality occurs, it is an object of the present invention to provide a chromatographic scanner capable of correcting such locality and achieving high-speed scanning of a light beam.

(Means for Solving the Problems) The chromatographic scanner of the present invention will be described with reference to FIG. 1 and FIG. 2 showing an embodiment. On the exit slit 10 of the spectroscope 8 or at its image forming position or in the vicinity thereof, A rotary slit disk 12 having slits 14-1, ... whose distance from the center changes depending on the angle is installed in a state where the longitudinal direction of the exit slit 10 is the radial direction of this slit disk 12, Rotate the disk 12 to slit 14
The detection signals from the means 20, 36 for detecting the moving light beam irradiation position and the detector 28 of the measuring light while moving the light beam irradiation position by passing through the hole at the intersection of 1, ... And the exit slit 10. When the sample plate 26 is measured using the data 36 for each light beam irradiation position, which has a means 36 for storing the light beam irradiation position in relation to each other, and which is measured in a state where the sample plate 26 is not present at the measurement position and is stored in the storage means 36. It is provided with means 36 for performing a locality correction calculation on the detection signal from the detector 28.

(Operation) When the slit disk 12 rotates, the hole at the intersection of the exit slit 10 and the slits 14-1, ... Of the slit disk 12 moves along the longitudinal direction of the exit slit 10. Along with this, the luminous flux irradiation position on the sample surface 26 also moves.

The position of the speed of light on the sample surface 26 corresponds to the rotation angle Θ of the slit disk 12, and the rotation angle Θ depends on the number of driving pulses of the pulse motor 16 that drives it or the driving of the synchronous motor. Detected by time.

Then, in the CPU 36, the sample surface (plate) 26 is placed at the measurement position.
Using the detection signal data from the detector 28 for each luminous flux irradiation position which is measured and stored in the absence of the condition, the locality correction calculation is performed on the detection signal from the detector 28 when measuring the sample surface 26.

(Embodiment) FIG. 1 shows an optical system and a signal processing system of a chromatographic scanner according to an embodiment. Reference numerals 2 and 4 are light sources, and two types are easily provided to cover a wide measurement wavelength range, and for example, a xenon lamp, a tungsten lamp or a deuterium lamp is used. Reference numeral 6 denotes a spherical mirror for selecting a light source, which is the light source 2 or 4
The speed of light from is incident on the spectroscope 8. Reference numeral 10 denotes an exit slit of the spectroscope 8 from which the separated light flux is emitted.

A slit disk 12 is installed on the exit slit 10 of the spectroscope 8. As shown in FIG. 2, this slit disk 12 is provided with, for example, three slits 14-1, 14-2, 14-3, and each slit 14-1, 14-2, 14-3 is The distance R from the center O is formed in a shape that changes according to R = R ₀ + kΘ depending on the rotation angle Θ. Here, R ₀ is the distance between the innermost position of the slits 14-1, 14-2, 14-3 and the center O. The center O of the slit disk 12 is fixed to the rotating shaft of the pulse motor 16 so that the slit disk 12 can be driven straight.

As shown in FIG. 2, the relationship between the slit disk 12 and the exit slit 10 of the spectroscope 8 is such that the slit disk 12 is positioned in the radial direction of the slit disk 12 in the longitudinal direction of the exit slit 10. Has been done. The hole 18 on the slit disk 12 is for detecting the origin of the slits 14-1, 14-2, 14-3, and is detected by the detector 20 shown in FIG.

Exit slit 10 of spectroscope 8 and slit of slit disk 12
The light flux that has passed through the hole at the position intersecting with 14-1, 14-2 or 14-3 is irradiated onto the sample plate 26 via the spherical mirror 22 and the plane mirror 24. The sample plate 26 is supported on a stage (not shown) movable in the X and Y directions.

Reference numeral 28 denotes a detector that detects the light flux that has passed through the sample plate 26. For example, a photomultiplier tube is used, and the transmitted light flux is received through a diffusion plate 29 such as a polytetrafluoroethylene plate. 30 is a preamplifier (preamplifier) for amplifying the detection signal of the detector 28, 32 is a LOG amplifier (logarithmic conversion amplifier) for converting the output signal of the preamplifier 30 into a logarithmic value, and 34 is L
A / D that converts the output signal of the OG amplifier 32 into a digital signal
A converter, 36 is a CPU that inputs the digital signal to calculate the absorbance and controls the rotation of the pulse motor 16.
Is. The CPU also determines the luminous flux irradiation position on the sample plate 26 from the signal from the detector 20 for detecting the origin of the slit disk 12 and the rotation angle of the slit disk 12, and then performs the locality correction calculation.

Now, assuming that the slit disk 12 is viewed in the direction of the arrow from the state shown in FIG. 2, the outlet slit 10 and the slit 14
The hole at the intersection with -1 moves away from the center O of the slit disk 12 along the longitudinal direction of the exit slit 10. Along with this, the light beam irradiation position on the sample plate 26 is (a) → (b) →
It moves like (c) → …… → (d). When the slit disk 12 further rotates and the next slit 14-2 intersects with the exit slit 10, the light beam irradiation position on the sample plate 26 returns to FIG. 3 (a) again. And one slit 14-
Each time the scanning on the sample plate 26 by 1, 14-2 or 14-3 is completed, the sample plate 26 is moved in the Y direction by a predetermined pitch by the movement of the stage. When the scanning of one lane in the Y direction is completed in this way, the stage is moved in the X direction to scan the next lane.

Next, the locality correction in this embodiment will be described with reference to FIGS. 4 and 5.

First, as the initialization routine, the stage is moved to create a state without a sample plate, and in that state, the slit disk 12 is rotated to absorb the light a ₁ , a ₂ for each position X ₁ , X ₂ , X ₃ , ... Xn. , a ₃ , ... An are measured by the detector 28 and stored in association with the rotation angle (beam irradiation position) of the slit disk 12. The positions X ₁ , X ₂ , ..., Xn are detected by the CPU 36 from the signal from the origin detection detector 20 and the rotation angle of the pulse motor 16, that is, the number of drive pulses of the pulse motor 16.

Next, the stage is moved to move the sample plate 26 to the measurement position, and measurement is performed to perform locality correction.
The process is performed as shown in FIG. When the measurement is performed at a certain point Xi on the sample plate 26, the CPU 36 reads the absorbance Ai from the A / D converter 34 (step S1), and the absorbance ai at the position Xi measured and stored by the initialization routine is stored.
Is called to calculate the difference Ai−ai between the two absorbances (steps S2 and S3). This is the locality correction.

After storing the calculated value in the memory (step S4), one drive pulse (or a predetermined number of pulses) is sent to the pulse motor 16 to move the luminous flux irradiation position on the sample plate 26 (step S5). Thereafter, the locality correction at each measurement point is performed in the same manner.

In addition, in the above embodiment, the hole for detecting the slit origin
18 may be one as in the embodiment, or slit 14-
You may provide in the number of 1-14-3.

Further, the slits on the slit disk 12 are not limited to those in the embodiment. For example, the number of slits may be four or more, or the number of slits may be one, and the pulse motor 16 may be reciprocally rotated to reciprocate the one slit. It is also possible to reciprocate the hole along the longitudinal direction of the outlet slit 10. The shape of the slit of the slit disk 12 is not limited to that of the embodiment, and any shape may be used as long as the distance from the center changes so as to correspond to the rotation angle Θ.

In FIG. 1, the slit disk 12 is installed on the exit slit 10, but in such a state, for example, the exit slit 10
This can be realized by forming an image in the X direction on top and forming an image in the Y direction on the slit disk 12. Further, the slit disk 12 may be installed at the position where the exit slit 10 of the spectroscope 8 is imaged. Further, the slit disk 12 may be installed in the vicinity of a position that is not strictly limited to the exit slit position and is separated as far as practically does not interfere.

The slit disk 12 can be driven by a synchronous motor instead of the pulse motor. In that case, the rotation angle of the slit disk 12 can be detected from the time from the origin detection.

If the slit disk is directly driven by a pulse motor or a synchronous motor, the durability is high, and since the light beam irradiation position on the sample surface can be detected from the rotation angle of the motor, the locality correction becomes easy.

(Effect of the Invention) According to the present invention, since the light beam can be scanned on the sample plate by rotating the slit disk in one direction or in the reciprocating direction, the structure is simple and the high speed is achieved. Scanning is possible.

Further, the locality correction corrects the variation in sensitivity due to the position on the light receiving surface of the detector, which improves the measurement accuracy.

Further, the slit disc can be made smaller in size according to the slit type of the slit disc in the present invention.

[Brief description of drawings]

FIG. 1 is a diagram showing an embodiment of an optical system as a schematic perspective view and a signal processing system as a block diagram. 2 is a plan view mainly showing a slit disk, FIG. 3 is a view showing a movement state of a light beam irradiation position on a sample surface in the same embodiment, and FIGS. 4 and 5 are locality correction in the same embodiment. 3 is a flowchart showing the operation of FIG. 8 ... Spectrometer, 10 ... Exit slit, 12 ... Slit disk, 14-1, 14-2, 14-3 ... Slit, 16 ... Pulse motor, 18 ... Origin detection hole, 20 ... ... Detector for origin detection.

Claims (1)

[Claims] 1. A rotary slit disk having a slit on the exit slit of the spectroscope, at its image forming position, or in the vicinity thereof, the distance from the center of which changes depending on the angle. Installed in the radial direction of the disk, rotate this slit disk to move the light beam from the spectroscope through the hole at the intersection of both slits and move the light beam irradiation position at the measurement position. A means for detecting the luminous flux irradiation position and a means for storing the detection signal from the measurement photodetector in association with the luminous flux irradiation position, and measuring with no sample plate at the measuring position and storing the luminous flux in the storage means. A chromatograph equipped with means for performing a locality correction calculation for a detection signal from a measurement photodetector at the time of measuring a sample plate using data for each position. Scanner.