JP2010181234A - Autoanalyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To realize an autoanalyzer capable of accurately confirming whether the magnification and target of an optical system are shifted from a focus position at a high speed. <P>SOLUTION: A plurality of chips 2 are successively fed to the place positioned under a lens 3 by a chip feeding part 1. The substance to be measured of each of the chips 2 is imaged by an imaging element 4 through the lens 3 to be stored in a data memory 5. The data stored in the data memory 5 are supplied to the fast Fourier transform part 71 of a magnification and focus position confirming part 7. The data subjected to Fourier transform by the fast Fourier transform part 71 are supplied to a magnification-focus position determining part 72, and imaging magnification and the focus position are determined. The determined result is supplied to a display part 8 to be displayed. A control part 6 analyzes the substance to be measured on the basis of the data stored in the data memory 5. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an automatic analyzer that performs quantitative measurement of light emission of a minute amount of fluorescence or phosphorescence by bioluminescence, chemiluminescence, or electrochemiluminescence.

  A technique is known in which a labeling substance is attached to a substance that specifically binds to a substance to be measured, and the labeling substance is measured qualitatively and quantitatively to perform qualitative and quantitative analysis of the substance to be measured.

  This technique has been applied to a method for determining the presence or absence of an antigen, antibody, or enzyme in a biological sample, or measuring the presence or absence of a gene having a specific sequence. In the past, there were times when radioactive substances were used as labels, but there are problems with easy handling due to radioactivity problems, etc., and in recent years it has become common to use substances that emit light as labels. is there.

  In particular, by immobilizing a substance that specifically binds to the substance to be measured in a specific area on the substrate surface, the substance to be measured is captured in the specific area, and the substance to be measured captured on the substrate surface is labeled. There is known a method for quantifying the amount of a labeling substance existing in a specific region by projecting light from the substance onto an image sensor by an imaging optical system.

  By this method, a high S / N can be realized by locally limiting the inspection region and increasing the density by integration of a specific region of the substance to be measured, thereby enabling a minute measurement.

  Furthermore, qualitative and quantitative analysis of multiple types of substances to be measured can be performed by immobilizing substances that specifically bind to different substances to be measured in multiple areas on the same substrate or in multiple substrates, and imaging multiple areas at once. Can be done. Since a plurality of items can be inspected by a single measurement, the amount of the sample and the amount of the reaction reagent can be reduced, and the output of the inspection result can be speeded up.

  In the imaging system of the automatic analyzer, the light from the marker present on the imaging surface is transmitted to the imaging device by the imaging optical system, but the amount of light projected per unit imaging device depends on the optical magnification and the focus position. To do.

  That is, when the optical magnification is high, the amount of light projected per unit image sensor decreases, and when the optical magnification is low, the amount of light projected per unit image sensor increases.

  In addition, even when deviating from the focus position, the image spreads and the amount of light projected per unit image sensor decreases.

  As described above, the amount of signal obtained from the image sensor varies depending on the difference in optical magnification and focus position. Therefore, in order to guarantee the reliability of the analysis result, an image is captured at a predetermined magnification and focus position in the analyzer. It is necessary to confirm whether it is.

  Such confirmation is more important as the amount of the substance to be measured is extremely small.

  A telecentric optical system is known as an optical system in which the optical magnification is constant within the depth of field and the fluctuation is small. In the telecentric lens, even when the object plane fluctuates from the focus plane, the magnification fluctuation is small because the principal ray position fluctuation on the imaging plane is small.

  In addition, there exists a technique described in patent document 1 as a related well-known technique. The technique described in Patent Document 1 is intended to shorten the manufacturing time and reduce the manufacturing cost, and is obtained by transferring the cells while maintaining the mutual position of each cell on the chip by direct contact with the host DNA chip. A replicated DNA chip substrate.

JP 2003-28865 A

  Although the above-described telecentric optical system has a small change in magnification, blurring occurs when the object plane deviates from the focus position. Telecentric optical systems are used in optical systems for inspection devices because the magnification and focus are relatively difficult to change. However, the aperture ratio is relatively low, resulting in an expensive optical system. It is not suitable for realizing possible bright optical systems.

  On the other hand, in other optical systems such as a macro lens that can be manufactured relatively inexpensively, the optical magnification is generally determined by the relationship between the distance between the lens and the object, and the distance between the lens and the image sensor, and the object plane varies from the focus position. In such a case, the optical magnification fluctuates and the image is blurred.

  Compared to the above optical system, both the magnification and focus position are more likely to fluctuate.However, the longer the back focus and working distance, the longer the object plane will change from the focus position within the depth of field. It is also possible to make an optical system in which the deviation of the light beam is within the spatial resolution of the image sensor.

  In any optical system, if the object plane and imaging plane deviate from the focus position, it is unavoidable to generate blur. Even in the case where the object plane or the image sensor surface fluctuates from the focus position and the image is blurred, it can be made constant within the depth of field of the imaging system.

  In this way, the inherent characteristics of the optical system itself are the blur due to the shift of the focus position and the fluctuation of the magnification. Therefore, the magnification is constant and not blurred in the device that performs microanalysis. Must be confirmed by some method to ensure the reliability of the analysis results.

  In the prior art, magnification confirmation is performed by imaging an object of a known length such as a standard scale and comparing the actual size of the object itself such as the outer shape with the captured image size. In the case of this method, special image processing is necessary to extract the contour of the object and to specify the principal ray position.

  For confirmation of the focus position, there is known a method of confirming by differential calculation of a captured image of the inspection chip without using a standard scale, but the magnification cannot be confirmed at the same time.

  An object of the present invention is to realize an automatic analyzer capable of confirming at high speed and accurately whether the magnification of an optical system and whether an object is deviated from a focus position.

  In order to achieve the above object, the present invention is configured as follows.

  In an automatic analyzer having an imaging optical system for imaging a substance to be measured, and an arithmetic control unit for controlling the imaging optical system and analyzing the substance to be measured based on an image captured by the imaging optical system, at least Image information of a plurality of confirmation substances arranged on the substrate imaged by the imaging optical system, and a plurality of confirmation substances that do not contain the substance to be measured arranged at a constant distance at equal intervals And calculating the magnification of the imaging optical system based on the calculated cycle information and determining whether the focus position with respect to the substance to be measured is appropriate. Prepare.

It is possible to realize an automatic analyzer that can quickly and accurately confirm the magnification of the optical system and whether the object is displaced from the focus position.
An automatic analyzer capable of measurement can be realized.

It is a schematic operation | movement block diagram in the 1st Example of this invention. It is explanatory drawing of the 1st Example of this invention. It is explanatory drawing of the 2nd Example of this invention.

  Embodiments of the present invention will be described below with reference to the accompanying drawings.

  FIG. 1 is a schematic operation block diagram in the first embodiment of the present invention. In FIG. 1, a chip (substrate) 2 is disposed on a chip transfer unit 1. A plurality of chips 2 are sequentially transferred to a position located under the lens 3 by the chip transfer unit 1.

  The substance to be measured on the chip 2 is imaged by the image sensor 4 through the lens 3 and stored in the data memory 5. The data stored in the data memory 5 is supplied to the fast Fourier transform unit 71 of the magnification and focus position determination unit 7. The data Fourier-transformed by the fast Fourier transform unit 71 is supplied to the magnification / focus position determination unit 72 to determine the imaging magnification and the focus position. Then, the determination result is supplied to the display unit 8 and displayed.

  The arithmetic control unit 6 analyzes (analyzes) the substance to be measured based on the data stored in the data memory 5. The analysis result is displayed on the display unit 8 or the like. Further, the arithmetic control unit 6 controls the operations of the lens 3, the image sensor 4, the data memory 5, the magnification and focus position confirmation unit 7, and adjusts the magnification and the focus position.

  FIG. 2 is an explanatory diagram of the first embodiment of the present invention. The first embodiment of the present invention is an example of imaging a substrate having a periodic pattern for magnification measurement on a substrate.

  2A, the actual center positions of the objects to be measured 2a to 2d are arranged on the chip 2 so as to have an equal interval y. And the area | region containing the some to-be-measured substances 2a-2d of the chip | tip 2 is equidistant Y in the image obtained with the imaging optical system of the optical magnification A. FIG.

  The areas 2a to 2d including the substance to be measured are made of a substance or a structure that can be discriminated by fluorescence or reflected light from the fluorescent label, and are in the same focal plane as the substance to be measured and include the substance to be measured. Anything can be used as long as it can be identified.

  The analysis region 20 including the plurality of measured regions 2a to 2d is arranged so that the actual center positions are at equal intervals y. The interval y is arbitrary if the regions containing the measured substances do not overlap each other. The size of

  In the obtained optical image, a spatial periodic structure (positional periodic information) is obtained in a region including the region 20 including the plurality of substances to be measured 2a to 2d.

  As one method of spatial frequency analysis for obtaining a periodic structure of space, a fast Fourier transform can be used. The number of regions including the substance to be measured used for the analysis can be from two, but the larger the number, the better. In the example of FIG. 2, for the sake of explanation, the analysis result in the direction in which the regions including the substance to be measured are arranged at equal intervals is shown. That is, the luminance data in the direction aligned at equal intervals is calculated, and the calculated luminance data is fast Fourier transformed by the fast Fourier transform unit 71.

  FIG. 2B is a graph of the result of the fast Fourier transform, where the vertical axis indicates the luminance intensity and the horizontal axis indicates the spatial frequency.

  In FIG. 2B, the first peak is observed at the position of the spatial frequency 1 / Y. Further, the peak at the high spatial frequency 1 / Y observed in the focus image obtained when the object position is at the focus position is not observed in the blurred image 30 obtained when the object position deviates from the focus position. .

  The optical magnification A can be confirmed by A = Y / y from the first peak position 1 / Y by obtaining the periodic structure of the space of the obtained image using the above events. Whether or not the object is in the focus position is determined by whether a peak is observed at a high spatial frequency 1 / Y or a peak is observed at a high spatial frequency of about 1 / Y ′. Can do.

  This determination is performed by the magnification / focus position determination unit 72, and the determination result is displayed on the display unit 8.

  As described above, according to the first embodiment of the present invention, the luminance data of the region 20 including the plurality of substances to be measured 2a to 2d on the chip 2 is subjected to the fast Fourier transform, and the object to be measured is focused from the peak position. The optical magnification A is calculated from the interval Y between the measurement target objects in the optical image and the set actual interval y and displayed.

  Therefore, it is possible to realize an automatic analyzer capable of confirming at high speed and accurately whether the magnification of the optical system and whether the object is displaced from the focus position.

  Further, complicated object position and contour extraction processing is not required, and the image processing load on the apparatus is reduced.

  In addition, an expensive standard scale manufactured with high accuracy is not required, and inspection can be performed at a low price.

  Furthermore, since the reliability can be ensured, an automatic analyzer capable of minute measurement can be realized by a general and inexpensive optical system that is worried about fluctuations in magnification and focus position.

  FIG. 3 is an explanatory diagram of the second embodiment of the present invention. Since the apparatus configuration is the same as that shown in FIG. 1, its illustration and detailed description are omitted.

  In the second embodiment, a plurality of regions (confirmation substance regions) 22 arranged in advance at equal intervals y are formed on the chip 2. The plurality of regions 22 do not include the substance to be measured, and the substance to be measured is disposed in the region 21. A plurality of substances to be measured are arranged at the same interval as the region 22. The plurality of areas 22 are analysis areas 23 for magnification calculation and focus position determination.

  In FIG. 3A, regions 22 not including a plurality of substances to be measured arranged so that the center positions are at equal intervals y are equally spaced in an image obtained by an imaging optical system with an optical magnification A. Y. The region (substance region for confirmation) 22 that does not contain the substance to be measured is made of a substance or structure that can be discriminated by fluorescence or reflected light from the fluorescent label, and is in the same focal plane as the region 21 containing the substance to be measured. It doesn't matter.

  A plurality of regions 22 that do not include a region to be measured and the center positions are arranged at equal intervals y. However, y may have any size as long as the regions 22 that do not include a substance to be measured do not overlap each other. In this image, a periodic structure of the space is obtained in a region 23 that includes a region 22 that does not include a plurality of substances to be measured.

  As one method of spatial frequency analysis for obtaining the periodic structure of space, fast Fourier transform can be used as in the first embodiment. The number of the regions 22 that do not include the substance to be measured used for analysis can be two, but the larger the number, the better. In the example shown in FIG. 3B, for the purpose of explanation, an analysis result is shown in the direction in which the regions 22 not including the substance to be measured are arranged at equal intervals.

  In FIG. 3B, the first peak is observed at the position of the spatial frequency 1 / Y. In addition, the peak at the high spatial frequency 1 / Y observed in the focus image obtained when the object position is at the focus position is not observed in the blurred image obtained when the object position deviates from the focus position. .

  The optical magnification A can be confirmed by A = Y / y based on the first peak position 1 / Y by obtaining the periodic structure of the space of the obtained image using the above events. Whether or not the object is in the focus position is determined by whether a peak is observed at a high spatial frequency 1 / Y or a peak is observed at a high spatial frequency of about 1 / Y ′. Can do.

  In the second embodiment of the present invention, the same effect as that of the first embodiment can be obtained.

  As a third embodiment of the present invention, the chip 2 in the second embodiment does not include the substance 21 to be measured, and only the confirmation substance region 22 is arranged, and the chip 2 dedicated for determining magnification and focus position is used. This is an example of calculating the magnification and determining the focus position.

  In the third embodiment, the same effects as those of the first and second embodiments can be obtained.

  DESCRIPTION OF SYMBOLS 1 ... Chip conveyance part, 2 ... Chip | tip, 2a-2d ... Area | region containing to-be-measured substance, 3 ... Lens, 4 ... Image sensor, 5 ... Data memory, 6 ... Control unit (analysis unit), 7 ... Magnification and focus position determination unit, 8 ... Display unit, 20, 21 ... Analysis region, 22 ... Area not including the substance to be measured, 30 ... -Blurred image, 31 ... Focus image, 71 ... Fast Fourier transform unit, 72 ... Optical magnification calculation / Focus position determination unit

Claims (9)

In an automatic analyzer having an imaging optical system for imaging a substance to be measured, and an arithmetic control unit that controls the imaging optical system and analyzes the substance to be measured based on an image captured by the imaging optical system,
A substrate on which a plurality of substances to be measured are arranged at regular intervals and at equal intervals;
Positional period information is calculated from image information of a plurality of substances to be measured arranged on the substrate imaged by the imaging optical system, and a magnification of the imaging optical system is calculated based on the calculated period information. And a magnification and focus position determination unit for determining whether or not the focus position for the substance to be measured is appropriate,
An automatic analyzer characterized by comprising.   2. The automatic analyzer according to claim 1, further comprising a display unit that displays whether the magnification and focus position of the imaging optical system determined by the magnification and focus position determination unit are appropriate.   The automatic analyzer according to claim 2, wherein the magnification and focus position determination unit divides the fixed distance of the substrate by an interval between substances to be measured in an image captured by the image sensor, The magnification is calculated, and the positional period information is frequency information obtained by Fourier transforming the relationship between the luminance and position of the image, and it is determined whether or not the focus position is appropriate based on the peak of the waveform of the frequency information. The automatic analyzer characterized by doing. In an automatic analyzer having an imaging optical system for imaging a substance to be measured, and an arithmetic control unit that controls the imaging optical system and analyzes the substance to be measured based on an image captured by the imaging optical system,
A plurality of substances to be measured are arranged at regular intervals at regular intervals, and a plurality of confirmation substances that do not contain the substance to be measured are arranged at regular distances at the regular intervals, and a substrate,
The positional periodic information is calculated from the image information of the plurality of confirmation substances arranged on the substrate imaged by the imaging optical system, and the magnification of the imaging optical system is calculated based on the calculated periodic information. And a magnification and focus position determination unit for determining whether or not the focus position with respect to the confirmation substance is appropriate,
An automatic analyzer characterized by comprising.   5. The automatic analyzer according to claim 4, further comprising a display unit that displays whether or not the magnification and focus position of the imaging optical system determined by the magnification and focus position determination unit are appropriate.   The automatic analyzer according to claim 5, wherein the magnification and focus position determination unit divides the certain distance of the substrate by an interval between the confirmation substances in the image captured by the image sensor, The magnification is calculated, and the positional period information is frequency information obtained by Fourier transforming the relationship between the luminance and position of the image, and it is determined whether or not the focus position is appropriate based on the peak of the waveform of the frequency information. The automatic analyzer characterized by doing. In an automatic analyzer having an imaging optical system for imaging a substance to be measured, and an arithmetic control unit that controls the imaging optical system and analyzes the substance to be measured based on an image captured by the imaging optical system,
A substrate on which a plurality of confirmation substances that do not contain a substance to be measured are arranged at regular intervals and at equal intervals;
The positional periodic information is calculated from the image information of the plurality of confirmation substances arranged on the substrate imaged by the imaging optical system, and the magnification of the imaging optical system is calculated based on the calculated periodic information. And a magnification and focus position determination unit for determining whether or not the focus position with respect to the confirmation substance is appropriate,
An automatic analyzer characterized by comprising.   8. The automatic analyzer according to claim 7, further comprising a display unit that displays whether or not the magnification and focus position of the imaging optical system determined by the magnification and focus position determination unit are appropriate.   The automatic analyzer according to claim 8, wherein the magnification and focus position determination unit divides the fixed distance of the substrate by an interval between confirmation substances in an image captured by the image sensor, The magnification is calculated, and the positional period information is frequency information obtained by Fourier transforming the relationship between the luminance and position of the image, and it is determined whether or not the focus position is appropriate based on the peak of the waveform of the frequency information. The automatic analyzer characterized by doing.

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