JP3706450B2 - Compound microscope - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a composite microscope.
[0002]
[Prior art and its drawbacks]
For example, a laser microscope or an X-ray microscope has been conventionally used to measure and analyze a sample that cannot be observed with the naked eye, such as a microorganism.
[0003]
On the other hand, as a device for measuring a two-dimensional distribution of a chemical substance that changes due to the activity of a microorganism, there is a two-dimensional concentration distribution measuring device composed of a LAPS sensor (Light-Addressable Potentometric Sensor). This two-dimensional concentration distribution measuring device was recently developed to two-dimensionally measure the pH of a substance dissolved in a liquid or a liquid soaked in a substance. For example, Jpn. J. et al. Appl. Phys. Vol. 33 (1994) pp L394-L397, a semiconductor substrate on which a sensor surface is formed is scanned with an appropriate light, and a photocurrent induced in the semiconductor substrate by this scan is taken out to perform measurement. be able to.
[0004]
The concentration distribution of the dissolved substance is measured by inserting or contacting the sensor surface directly with the target substance to be measured. The obtained data is output as a two-dimensional density distribution image by computer processing. Not only the concentration distribution at a certain time but also the state of the change can be tracked in real time.
[0005]
The applicant of the present application has applied for a patent on the device used for measuring the two-dimensional distribution of the ion concentration such as pH as an “optical scanning device” dated February 4, 1995. Application No. 7-39114).
[0006]
Originally, in research on microorganisms and the like, it is particularly important to obtain characteristics of exactly the same object under the same conditions. By using the optical scanning device, metabolism of living samples such as microorganisms can be measured. However, since the activity of microorganisms varies depending on temperature and other conditions, analysis is performed in comparison with the measurement results of other microscope devices. Can not. Thus, a plurality of measurements cannot be performed under the same conditions. Also, even if a two-dimensional measurement is performed on a fine object and a plurality of microscope observation results are overlapped with the same distribution and position, the sample conditions may not always be the same. In some cases, the analysis results are often meaningless.
[0007]
The present invention has been made in consideration of the above-described matters, and an object of the present invention is to provide a composite microscope capable of performing composite measurements.
[0008]
[Means for Solving the Problems]
In order to achieve the above object, in the present invention, a combined function is obtained by combining a current light scanning chemical microscope or potential light scanning chemical microscope with a laser microscope or an X-ray microscope.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
In the first composite microscope of the present invention, a sensor part made of a photoconductive layer showing conductivity is formed only on a part of the light-transmitting substrate part that is directly irradiated with light, and the sensor part is formed on the sensor part. The counter electrode and the reference electrode are brought into contact with the sample arranged so as to be in contact with each other, a voltage is applied between the counter electrode and the substrate portion, and irradiation is performed while scanning light from the other surface of the substrate portion. The present invention is characterized in that a current-type optical scanning chemical microscope in which a current change caused by the above is taken out from a light-transmitting substrate portion and a laser microscope or an X-ray microscope are combined.
[0010]
And the 2nd compound microscope of this invention forms the sensor part which consists of an ion response film on one side of a semiconductor substrate via an insulating layer, and sets a counter electrode to the sample arranged so as to contact this sensor part. A reference electrode is brought into contact, a voltage is applied between the counter electrode and the semiconductor substrate, and irradiation is performed while scanning light from the other surface of the semiconductor substrate, and a photocurrent generated in the irradiated portion is taken out of the semiconductor substrate. The potential-type optical scanning chemical microscope and a laser microscope or an X-ray microscope are combined.
[0011]
【Example】
FIG. 1 schematically shows the configuration of a first composite microscope of the present invention. In FIG. 1, A is a current type optical scanning chemical microscope, which is configured as follows. That is, reference numeral 1 denotes a light-transmitting substrate portion, for example, a light-transmitting substrate (hereinafter referred to as a transparent substrate) 2 and light transmission provided in close contact with one surface (upper surface in the illustrated example) of the transparent substrate 2. Electrode 3 (hereinafter referred to as a transparent electrode). As the transparent substrate 2, for example, a glass plate is used, and as the transparent electrode 3, for example, ITO (transparent conductive film) obtained by mixing In ₂ O ₃ and SnO ₂ at an appropriate ratio is used.
[0012]
4 is a photoconductive layer as a sensor portion formed on the upper surface of the transparent electrode 3, and this photoconductive layer 4 exhibits conductivity only in a portion directly irradiated with light. For example, amorphous silicon ( It is formed by treating α-Si) at a temperature of 300 to 350 ° C.
[0013]
Reference numeral 5 denotes a side wall which is erected so as to be in close contact with the upper surface of the photoconductive layer 4 and forms a sample container 6 together with the photoconductive layer 4 and is made of an appropriate resin material.
[0014]
Reference numeral 7 denotes a measurement sample including, for example, the cells 7a, and is placed on the flat sensor unit 4 in a gel state.
[0015]
Reference numerals 8 and 9 denote a counter electrode and a reference electrode arranged so as to be in contact with the measurement sample 7, and each of them is composed of, for example, a Pt electrode and an Ag—Cl electrode.
[0016]
Reference numeral 10 denotes a light source that irradiates the laser beam 11 onto the substrate unit 1 from the lower surface side, and is scanned in the X and Y directions by a scanning mechanism (not shown) controlled by the scanning control device 12.
[0017]
Reference numeral 13 denotes a control box, which includes a potentiostat 14 that applies a voltage between the counter electrode 8 and the transparent electrode 3 and extracts a signal obtained at that time as a current signal, and an interface board 15 that exchanges signals with the potentiostat 14. Become. The potentiostat 14 receives a stabilizing bias circuit 16, a current-voltage converter 17 that converts a current signal extracted from the transparent electrode 3 into a voltage signal, and a signal from the current-voltage converter 17. And an operational amplifier circuit 18.
[0018]
Reference numeral 19 denotes a computer as a control / arithmetic unit that performs various controls and computations, and sends control signals to the scanning control device 12 and processes signals from the control box 13 as appropriate. The processing result is displayed on a display unit 19a such as a color display or stored in a memory (not shown).
B is a laser microscope, which is a known confocal type, and is provided so that the objective lens faces the measurement sample 7 provided on the upper surface of the sensor unit 4. The laser microscope B is controlled by the computer 19, and image data from the laser microscope B is sent to the computer 19 for appropriate signal processing and displayed on a display unit 19 b provided in parallel with the display unit 19 a, It is stored in the memory of the computer 19.
[0020]
In the composite microscope having the above configuration, the ion-sensitive dye is infiltrated into the measurement sample 7 to color the cells 7 a in the measurement sample 7, and this is placed on the sensor unit 4. Then, the ion concentration (for example, pH) distribution outside the cell 7a can be measured by the current-type optical scanning chemical microscope A, and the ion concentration distribution in the cell 7a can be measured by the laser microscope B. These measurement results are displayed on the display units 19a and 19b of the computer 19, respectively.
[0021]
In the above-described embodiment, the inside and outside of the cell 7a in the measurement sample 7 are measured simultaneously. However, the present invention is not limited to this and can be implemented with various modifications. Instead of the laser microscope B, an X-ray microscope may be used, and this may be combined with the current-type optical scanning chemical microscope A. For example, when a living organism is a measurement target, it is observed with an X-ray microscope in a living state, and the metabolic activity of the organism is measured in real time, so that an image of various ion concentration distributions, enzyme reactions or immunosensitive reactions can be obtained. Can be displayed.
[0022]
And when the said current type optical scanning chemical microscope A and X-ray microscope are combined, the thin film layer of an electronic device can also be used as a measurement sample. That is, while the X-ray microscope can display the distribution of elemental components formed in each layer, the current-type optical scanning chemical microscope A can detect the potential distribution and pinholes of the thin film.
[0023]
In any of the above-described embodiments, the current type optical scanning chemical microscope A is used. However, instead of this, a potential type optical scanning chemical microscope can be used. This will be described below with reference to FIG.
[0024]
FIG. 2 schematically shows the configuration of the second composite microscope of the present invention. In this embodiment, a potential light scanning chemical microscope A ′ is used. That is, 20 is a semiconductor substrate, which is made of, for example, a silicon substrate, and this semiconductor substrate 20 is electrically connected to the current-voltage converter 17 of the potentiostat 14 via the ohmic electrode 21.
[0025]
Reference numeral 22 denotes an insulating layer made of, for example, SiO ₂ provided in close contact with one surface (upper surface in the illustrated example) of the silicon substrate 20. Reference numeral 23 denotes an ion-responsive film as a sensor unit formed in close contact with the upper surface of the insulating layer 22, which is made of, for example, Si ₃ N ₄ and is configured to respond to hydrogen ions.
[0026]
Reference numeral 24 denotes a side wall which is erected so as to be in close contact with the upper surface of the ion response film 23 and which forms the sample holder 25 together with the ion response film 23 and is made of an appropriate resin material.
[0027]
Other configurations are the same as those shown in FIG. 1, and thus detailed description thereof is omitted. Since the operation in this embodiment is the same as that of the first invention, its detailed description is omitted.
[0028]
In addition, the device kit of the current-type optical scanning chemical microscope A and the potential-type optical scanning chemical microscope A ′ may be attached to the housing of the laser microscope B or the X-ray microscope. The whole structure is neat and can be handled easily.
[0029]
【The invention's effect】
The present invention is implemented in the form as described above, and has the following effects.
[0030]
(1) Observe and measure the same sample with two types of microscopes with different characteristics under the same processing conditions, and simultaneously analyze and measure the distribution, so analysis that cannot be analyzed with one conventional microscope is performed. I was able to do it.
[0031]
(2) There is no need to purchase expensive microscopes.
[0032]
(3) There is no need to make a sample many times.
[Brief description of the drawings]
FIG. 1 is a diagram schematically showing a configuration of a first composite microscope of the present invention.
FIG. 2 is a diagram schematically showing a configuration of a second composite microscope of the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Board | substrate part, 4 ... Sensor part, 7 ... Sample, 8 ... Counter electrode, 9 ... Comparison electrode, 20 ... Semiconductor substrate, 22 ... Insulating layer, 23 ... Sensor part, A ... Current type optical scanning chemical microscope, A ' ... potential type optical scanning chemical microscope, B ... laser microscope or X-ray microscope.

Claims (2)

A sensor part made of a photoconductive layer showing conductivity is formed only on a part of the light-transmitting substrate part that has been directly irradiated with light, and a counter electrode is placed on the sample placed in contact with the sensor part. The reference electrode is brought into contact, a voltage is applied between the counter electrode and the substrate portion, and irradiation is performed while scanning light from the other surface of the substrate portion. A composite microscope comprising a combination of a current-type optical scanning chemical microscope that can be taken out from a part and a laser microscope or an X-ray microscope. A sensor unit made of an ion-responsive film is formed on one surface of the semiconductor substrate via an insulating layer, and a counter electrode and a reference electrode are brought into contact with a sample arranged so as to be in contact with the sensor unit. A potential-type optical scanning chemical microscope that applies a voltage between and irradiates while scanning light from the other surface of the semiconductor substrate, and takes out the photocurrent generated in the irradiated portion from the semiconductor substrate; A composite microscope characterized by combining a laser microscope or an X-ray microscope.

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