JPS61262635A - Chromatographic scanner - Google Patents

Abstract

PURPOSE:To shorten the scanning time without lowering the accuracy of quantitative analysis, by automatically limiting a range performing highly accurate final scanning by rough scanning. CONSTITUTION:A specimen 3 such as a thin layer chromatography plate or an electrophoretic gel is held on a specimen stage 3 movable to both X- and Y-directions and irradiated with light from an optical system 2. The specimen stage 20 is controlled by a microcomputer MC12 through a drive mechanism and linear scanning being rough scanning and zigzag scanning being main scanning are performed. The reflected light from the specimen 3 is converted to an electric signal by a photomultiplier tube 4 and the transmitted light from the specimen 3 is converted to an electric signal by a photomultiplier tube 6 while both signals are amplified by an amplifier 8 and converted by an A/D converter 10 to be taken in MC12. MC12 detects the peak position at the time of rough scanning from the taken-in signal and the main scanning range performing zigzag scanning is limited only to a peak range containing a spot and data processing at the time of final scanning is performed to output the processed data to a printer 14 and CRT16.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to a chromatography scanner that irradiates light onto a sample such as a TLC (thin layer chromatography) plate or electrophoresis gel and performs quantitative determination using the reflected or transmitted light. It is related to.

(Prior Art) A TLC plate or an electrophoresis gel has a pattern that spreads discretely in a straight line on a sample.

Figure 8 is an example of a TLC plate, Figure 9 is an example of an electrophoresis gel, and 30.32 is a spot.

Conventional chromatography scanners perform quantification by scanning a pattern spread discretely on a straight line in the Y direction, such as on a TLC plate or electrophoresis gel.

The scanning start point of the zigzag scan was set at the end of the scanning range of the rectangle, the number of steps in the Y direction was kept constant, and the entire surface of the rectangle determined by the swing width of the zigzag was scanned as shown in Figure 10. .

(Problem to be Solved by the Invention) Although the spots on a sample are discrete, in conventional chromatography scanners, areas where there are no spots must be scanned in the same way as important areas where spots are present, especially for quantitative analysis. A huge amount of time is wasted on unnecessary parts, such as when performing integration to improve accuracy.

An object of the present invention is to provide a chromatography scanner that automatically searches for the position of a sample spot at high speed, thereby shortening scanning time without reducing quantitative accuracy.

(Means for Solving the Problems) In the present invention, as shown in FIG. 1, a mode switching section switches between a coarse scanning mode and a main scanning mode.

During rough scanning, the stage control unit causes the sample stage 20 to fix the X direction and perform linear scanning only in the Y direction. During this linear scanning, the peak processing section detects the peak position from the detection signal, and the peak position is stored in the memory 18.

During the main scan, the sample stage 20 performs a zigzag scan by the stage control section, but at this time, the stage control section performs a highly accurate zigzag scan at the peak position according to the peak position stored in the memory 18, and performs a highly accurate zigzag scan at the peak position. Then, the sample stage 20 is controlled so that the zigzag scan is omitted or the scan is performed roughly. The data processing section performs quantitative data processing from the detection signals during the main scan.

(Example) FIG. 2 represents one embodiment of the invention.

2 is an optical system that irradiates the sample 3 with light.

Sample 3 such as TLC plate or electrophoresis gel is X, Y
It is held on a sample stage 20 that can move in both directions. The sample stage 20 is a drive mechanism (not shown)
It is controlled by the microcomputer 12 via the microcomputer 12 to perform coarse linear scanning and main scanning zigzag scanning.

Reference numeral 4 denotes a photomultiplier tube as a photoelectric conversion unit that receives reflected light from the sample 3 and converts it into an electrical signal, and 6 refers to a photoelectron as a photoelectric conversion unit that receives transmitted light from the sample 3 and converts it into an electrical signal. It is a multiplier tube. The output signal of the photomultiplier tube 4 or 6 is amplified by an amplifier 8, converted into a digital signal by an A/D converter 10 serving as a signal converter, and then taken into a microcomputer 12.

The microcomputer 12 performs peak position detection during rough scanning and data processing during main scanning from the captured signals.

14 is a printer for displaying the results of data processing by the microcomputer 12, and 16 is a CRT.

The microcomputer 12 implements a data processing section, a peak processing section, a memory, a stage control section, and a mode switching section in the present invention.

Next, the operation of this embodiment will be explained with reference to FIGS. 2 to 7.

The sample stage 20 is positioned so that the irradiation position of the sample 3 by the optical system 2 is at the scanning start point (step S1).
. Set the measurement mode to coarse scanning mode (step S2
), whereby the sample stage 20 is controlled by the microcomputer 12 to perform linear scanning of the row of spots 30 without integration by fixing the X direction and increasing the feed in the Y direction, as shown in FIG. (Step S3).

This linear scanning causes the photomultiplier tube 4 or 6 to
The waveform of reflected light intensity or transmitted light intensity as shown in the figure is detected. The microcomputer 12 performs peak processing to obtain peak positions such as a peak start position 71w, a peak top position y2, and a peak end position y3 for each peak from this waveform (step S4). This can be done by differentiating and using the differential value.

Next, the peak range 71"" obtained by this peak processing
'y3 is expanded by an appropriate amount as shown in FIG.
6).

Next, position the irradiation position again to the scanning start point of the sample,
Set the measurement mode to main scanning mode (step S7)
. The microcomputer 12 reads out the first peak position from the memory (step S8), and the sample stage 20
is moved to the peak position, the original zigzag scanning is performed for the peak range AI shown in FIG. 7, and the detection signal is taken in and data processing is performed (step S9.5IO). This zigzag scanning and data processing is conventional.

When the zigzag scan of the first peak range A1 is completed, the microcomputer 12 reads out the next peak position and similarly performs data processing by zigzag scan.In this way, A+ v A2. A3.・
The zigzag scan of the peak range indicated by . . . is sequentially repeated until all peaks are completed (step 511).

Through the above processing, the range in which the zigzag scan is performed is limited to only the area where the spot is present, and the background area, which occupies most of the area, is excluded from the target of the zigzag scan, thereby shortening the scanning time.

In the above embodiment, the main scanning range for performing zigzag scanning is limited to only the peak range including the spot.

In areas other than the peak range, main scanning is not performed at all. However, for example, although zigzag scanning is performed in areas other than the peak range, the feed in the Y direction may be increased or the number of integrations may be reduced.

Such an operation can also shorten the scanning time.

(Effects of the Invention) The chromatography scanner of the present invention includes an optical system, a photoelectric conversion section, a signal conversion section, a sample stage whose drive is controlled by a microcomputer, and a data processing section implemented by the microcomputer.

It is equipped with a peak processing unit, memory, stage control unit, and mode switching unit, and operates to automatically limit the range of highly accurate main scanning based on a rough survey, reducing the scanning time in the background area where there are no spots. The scanning time can be significantly shortened without reducing quantitative accuracy.

[Brief explanation of drawings]

FIG. 1 is a block diagram showing the configuration of the present invention, FIG. 2 is a schematic diagram showing an embodiment of the present invention, FIG. 3 is a flowchart showing the operation of one embodiment, and FIG. 4 is a rough diagram of the same embodiment. FIG. 5 is a diagram showing a linear scan, and FIG. 5 is a diagram showing a detected waveform obtained by the linear scan. Figure 6 is a diagram showing the peak range stored in memory, and Figure 7 is a diagram showing the peak range stored in memory.
The figure shows the main scanning range in the same embodiment, and FIG.
FIG. 9 is a diagram showing an LC plate, FIG. 9 is a diagram showing an electrophoresis gel, and FIG. 10 is a diagram showing conventional zigzag scanning. 2...Optical system, 4.6...Photomultiplier tube, 10...A/D converter, 12...Microcomputer, 18...・・・
··memory. 20...Sample stage.

Claims (1)

[Claims] (1) An optical system that irradiates the sample with light, a photoelectric conversion unit that receives reflected light or transmitted light from the sample and converts it into an electrical signal, and a signal that converts the output signal from this photoelectric conversion unit into a digital signal. a converting section; a data processing section that processes and outputs the output signal of the signal converting section during main scanning; a peak processing section that detects a peak position from the output signal of the signal converting section during rough scanning; and this peak processing. a memory for storing the position of the peak detected in the section, a sample stage for moving the sample, and a sample stage for moving the sample according to a preset movement mode during rough scanning and according to the peak position stored in the memory during main scanning. A chromato scanner comprising: a stage control section that controls driving; and a mode switching section that switches operations of the data processing section, peak processing section, and stage control section between main scanning and coarse scanning.