JP2006162445A - Electrophoretic device, and photographic analytical system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an electrophoretic device capable of restraining conductivity from deteriorat due to bubbles, in the electrophoretic device constituted integrally with an electrical output device, and capable of attaining safety for an experiment person, and being easy to use, and an electrophoretic observation device that enables analysis after electrophoresis, at an accessible site, without utilizing a dark room. <P>SOLUTION: This electrophoretic device arranged with electrodes on its both side, arranged with gel for electrophoresis between the electrodes, and comprising a buffer solution arranged to immerse at least the gel for electrophoresis is provided with a partitioning plate, having an inclination arranged to be projected at least from the buffer solution in the vicinity of the electrodes. This electrophoretic observation device, having a casing storing the electrophoretic device to be used, formed of a material shielding external light, with at least one or more of opened side faces, and having a light-shielding cover for covering the opened side faces is provided with an opening in the side face of the light-shielding cover. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention mainly uses biomolecules such as DNA, RNA, and proteins, or mixtures of various low-molecular compounds and complexes having a charge, and electrophoresis used for separation analysis, purification, and recovery of these samples. The present invention relates to an apparatus and an imaging analysis system thereof.

As a conventional electrophoresis apparatus, an electrophoresis apparatus disclosed in Japanese Utility Model Publication No. 63-39639 is known. This electrophoresis apparatus is composed only of a fuse, a rectifier, an operation on / off switch, and an output changeover switch, and can supply a current corresponding to the impedance of the electrophoresis carrier.
Shoko Sho 63-39639 All of the electrophoresis methods (Medical and Dental Publishing Co., Ltd., 1983) In all of the electrophoresis methods (Medical and Dental Publishing Co., Ltd., 1983), electrodes are placed so as to sandwich the gel soaked in the buffer solution. In addition, a plate-like body having partitioning properties protruding from the buffer solution is disposed between the electrode and the gel, and a configuration in which the plate-like body and the bottom of the electrophoresis tank are used as an electrical passage is shown. .

However, since the inventive device is directly connected to a general-purpose AC power source such as an AC 100 V household power source, there is a high possibility that a large amount of current flows due to a rapid drop in impedance of the electrophoresis carrier or a short circuit of each part. Although this large amount of current is dangerous for the human body, the only consideration for electrical insulation is the disconnection of the built-in fuse and the protective functional form of the case housing the electronic circuit.
Such a simple protection function is not enough at present when electrophoresis experiments are increasing as part of education at high schools. In addition, if an unhandled person conducts an experiment, there is a high possibility that the device will fail, such as accidentally spilling liquid, so that it is difficult to make a configuration that does not cause failure or operations that may cause failure. Such an improvement in the feeling of use is required.

However, if the circuit configuration is complicated only to improve safety and usability, or if automation is promoted, the size and cost of the apparatus will increase. As a result, such devices can only be handled by specialized research institutes, and do not lead to expansion of educational applications or speeding up research by handling multiple units by a single researcher. Therefore, there is a need for an electrophoresis apparatus that is as simple as possible and safe and easy to operate.
The inventors have devised a simple and safe electrophoresis apparatus in Japanese Patent Application No. 2001-379255. In the electrophoresis apparatus according to the present invention, the lid of the electrophoresis tank is provided with a magnet, a reed switch is sandwiched between the connection part of the power supply for electrophoresis and the electrophoresis tank, and the reed switch is operated by the magnet of the lid. Without it, it has the safety of not energizing. In addition, the electrophoretic power switch is equipped with a magnet, and the reed switch in the power supply case is operated by magnetic force, so that the circuit part can be completely sealed in the case. A difficult configuration is realized.

However, the electrophoresis apparatus according to the present invention includes a problem that the operation may become unstable due to variations in performance of magnets used everywhere. In addition, since a reed switch for enhancing safety is attached to the electrophoresis tank, liquid may enter the switch part during washing or injection of a buffer solution, leading to failure. Furthermore, since the electrode wire is exposed at the bottom of the electrophoresis tank, it may be easily touched at the time of cleaning or the like, which may also cause a failure. For this reason, it is necessary to pay attention to handling, and it cannot be said that it can be used easily.

Therefore, the present invention is an extremely compact electrophoresis apparatus that takes advantage of the simplicity of the electric circuit disclosed in Japanese Utility Model Publication No. 63-39639, and has an excellent usability that anyone can easily handle, and is less likely to break down. The object is to devise an electrophoresis apparatus with improved safety.
The safety of the apparatus is ensured by a configuration that cannot be energized without a lid, but the lid and the switch provided in the electrophoresis tank are not interlocked as in the electrophoresis apparatus of Japanese Patent Application No. 2001-379255. And a switch provided in the power supply for electrophoresis. This makes it possible to eliminate switches that are sensitive to liquids from the electrophoresis tank, and dramatically reduce the possibility of failure. Furthermore, an extremely safe electrophoresis apparatus without any possibility of electric shock can be realized by adopting a switch configuration that cannot operate without a lid.

As the power supply switch for electrophoresis, a mechanical switch with little variation and stable operation is used. However, a waterproof structure is improved by devising the shape of the button and the case, and the structure is not easily damaged.
In addition, by devising the shape of the electrophoresis tank and lid, it is possible to transfer and dispose of the buffer solution safely and quickly, or to make it easy to check the interior even if the lid is cloudy. The feeling can be improved.

Furthermore, in an electrophoresis experiment, it is necessary to perform a series of procedures such as visualization of a sample after electrophoretic separation, photographing of a result, and analysis quickly and safely. In the present invention, not only the electrophoresis apparatus but also the photographing method and analysis method thereof are improved. Specifically, the irradiation device for visualizing the sample after electrophoresis, the observation imaging device for the visualized sample, and the software for analyzing the acquired image data are improved. With regard to the irradiation apparatus, by arranging small light sources, it is possible to perform irradiation with high uniformity while reducing the overall size, and further improve the irradiation uniformity by devising the shape of the light reflector. In the observation and photographing device, the overall durability and strength are achieved by configuring the casing with stainless steel material with excellent light-shielding properties, chemical resistance, and strength, and configuring both sides with dark curtains with excellent flexibility. A device that allows flexible and flexible internal processing while maintaining it. By making it possible to select and connect a fast-evolving digital camera as a photographic means, it has become a device with excellent future expandability. Finally, the analysis software adopts the most common multi-window format, allowing the main objects such as images and graphs to have parameters necessary for electrophoresis analysis as properties and to set the properties freely. By enhancing the interface, advanced analysis processing can be performed intuitively.

The present invention makes it possible to realize a safer and more efficient electrophoresis experiment, and allows an experimenter to smoothly supply necessary related-made instruments and consumables. In addition, the observation and photographing means after the electrophoresis experiment and the analysis means for the image data obtained therefrom are improved to realize a system that efficiently proceeds with a series of procedures related to the electrophoresis method.

BEST MODE FOR CARRYING OUT THE INVENTION

The electrophoresis apparatus and the imaging analysis system of the present invention will be described in order.
First, a cross-sectional view of the basic configuration of the electrophoresis apparatus of the present invention is shown in FIG.
The electrophoresis apparatus of the present invention is called a submarine type, and the electrophoresis buffer 2 is injected into the electrophoresis tank 1. The central part of the electrophoresis tank 1 is usually raised in a trapezoidal shape. In addition, by using a small amount of a supporting electrolyte such as a buffer solution used in the electrophoresis tank, it can be diverted to filter paper electrophoresis or cellulose acetate electrophoresis at a place where it rises in a trapezoidal shape. Electrodes 4 are stretched on both bottoms separated by the base 3. The electrode 4 is usually a platinum linear member. Each of the two electrodes 4 is connected to a migration power source, one of which is an anode and the other is a cathode.
An electrophoretic carrier 5 is installed on the platform 3. This electrophoresis carrier 5 is often a gel mainly composed of agarose in a submarine type electrophoresis apparatus. The electrophoresis carrier 5 is provided with a sample installation groove 6 in which the sample 7 is installed. A voltage is applied to the electrophoresis carrier 5 from the electrode 4 via the buffer solution 2, and the charged sample 7 is migrated. Each component contained in the sample 7 is separated in the electrophoresis carrier 5 according to a difference in molecular size or a difference in charged state.

  In addition, since a high voltage is usually applied in electrophoresis, an electrophoresis tank lid 8 is installed to prevent electric shock. The electrophoresis tank lid 8 also plays a role of suppressing evaporation of moisture in the buffer solution and preventing contamination such as dust from entering the electrophoresis tank. Further, as shown in FIG. 1A, the partition plate 8a may be installed above the electrode 4 as necessary. This is to cover at least the lower part so as not to obstruct energization from the electrode 4 but to prevent the electrode 4 from touching the electrode 4 easily from the upper part. The partition plate 8a also functions to prevent air bubbles generated from the electrodes 4 from flowing into the central portion of the electrophoresis tank 1. The configuration in which the partition plate 8a is arranged in a direction perpendicular to the bottom surface of the electrophoresis tank has been used for a long time, such as the one described in the column of non-patent literature. In order to influence electrophoresis, it is desirable that the partition plate 8a be a part that can be attached and detached as required, such as a plug-in type. Further, as shown in FIG. 1B, the partition plate 8b may be installed inclined from the vertical direction. This is a structure for preventing bubbles generated from the electrode 4 from adhering to the partition plate and remaining in the buffer solution 2 for migration. If bubbles remain, dissolution in the electrophoresis buffer 2 is promoted, and there is a possibility that electrophoresis will be affected by an increase in concentration in the solution.

Further, as shown in FIG. 1C, a bottom partition plate 9 may be installed. The bottom partition plate 9 is a plate erected upward from the bottom of the buffer solution tank of the electrophoresis tank 1 and is not necessarily vertical, but the upper limit of the bottom partition plate 9 is the partition plate (FIG. 1A). 1a or 8b of Fig. 1 (b) and Fig. 1 (c), so that the possibility of touching the electrode 4 is further reduced by the presence of the bottom partition plate 9. In addition, since the bottom partition plate 9 has a function of physically dividing the buffer solution tank, it is possible to cause discontinuity in the components and concentration of the buffer solution. The partition plates 8a and 8b and the bottom partition plate 9 do not necessarily have to be separated from each other, and may be incorporated into the migration tank as one component 8c having a cross section as shown in FIG. 1 (d). .
It is exemplified that the partition plate 8a is provided with a guide groove on the opposing inner surface in the migration tank, and is incorporated and installed in the migration tank so as to be inserted along the guide groove.
Other examples of the guide groove include those provided with two continuous convex portions in parallel or those provided with a rail-like body.
The inclination of the partition plate 8a is preferably 3 ° to 30 ° with respect to the side surface of the electrophoresis tank in that bubbles can be efficiently discharged to the outside, but may be inclined at other angles depending on the composition of the buffer solution. In some cases.
The partition plate 8a only needs to have at least a gap and a hole protruding from the buffer solution surface and penetrating inside the buffer solution. The shape of the partition plate 8a is not limited to these.

  In the electrophoresis experiment, different electrochemical reactions occur between the two electrodes, and generally different amounts of bubbles are generated. For example, an electrolysis reaction occurs in which oxygen is generated at the anode and hydrogen twice as much as that is generated at the cathode. Since the reaction is different between the two electrodes, the buffer solution in the vicinity of the two electrodes has different physical properties with the passage of time, affecting the electrophoresis. Even in the basic configuration as shown in FIG. 1, the circulation of the buffer solution occurs mainly due to the difference in the amount of generated bubbles, but the effect is small. Pumps that forcibly circulate the buffer solution for the purpose of making the physical properties of the buffer solution in the vicinity of both poles are also commercially available, but they are more expensive than the electrophoresis tank and require a lot of work to install, making it easy to use Not suitable for purpose. Therefore, as shown in FIG. 2, a method of configuring the inside for the purpose of circulating the buffer solution is considered. The base part of the electrophoresis tank 1 is used as a gel installation member 3a so that it can be attached and detached as necessary. There is a space between the lower surface of the gel installation member 3 a and the electrophoresis tank 1, and the space is filled with the buffer solution 2. In the basic configuration shown in FIG. 1, the buffer solution path between the electrodes exists only in the upper part of the gel. Therefore, the buffer solutions from both electrodes are pushed and exchanged here, but in the configuration shown in FIG. Since the liquid path exists, the buffer solution 2 is easily circulated.

That is, the buffer solution flows from the cathode (electrode with a large amount of bubble generation) 4a to the anode 4b (electrode with a small amount of bubble generation) at the top of the gel, and vice versa at the bottom. Circulate. Since the driving force for circulating the buffer solution is due to the generation of bubbles, the path constituting portion 3b extends from the gel installation member 3a toward the cathode 4a, and further, the bubble guiding portion 3c is formed above the cathode 4a. Yes. Due to the presence of the bubble guiding portion 3c, the bubbles generated from the cathode 4a are guided upward and are efficiently used as a driving force for circulating the buffer solution 2. The narrower the gap between the side surface of the electrophoresis tank 1 and the bubble guiding portion 3c, the more concentrated the flow of bubbles and the greater the driving force. However, since it affects the formation of the electric field, it cannot be made extremely narrow. To the extent. The path | route structure part 3b is for concentrating the driving force by a bubble on the force which draws in a buffer solution from an anode side. In addition, since the gel installation member 3a can be attached in the reverse direction as long as it is detachable, it can cope with the case where the amount of bubbles generated in both electrodes is different. Since the buffer solution bottom path 1A below the gel installation member 3a is energized, if this path is too large, conversely, consumption of the buffer solution is promoted. The diameter of the buffer solution bottom path 1A may be about 3 mm or less in height and about 400 mm ² or less in cross-sectional area, and the width on the cathode 4a side (side on which the buffer solution is sucked by bubbles) may be narrower than the anode 4b side.

FIG. 3 shows another embodiment of the present invention for safely and easily handling the electrophoretic tank 600A and the power supply unit 600 without touching the electrical connection portion between them from the outside.
Apart from the electrode terminal pin 602 connected to the power source, a safety pin 630 which is preferably made of a material having no conductivity is installed on the side of the electrophoresis tank.
It is preferable that the electrode terminal pin 602 and the safety pin 630 have approximately the same length and protrude in the direction of the electrophoresis tank. However, the safety pin 630 may differ depending on the position of the switch unit to be operated.
Furthermore, an L-shaped stopper 632 that can rotate around the shaft portion 632S is provided in the power supply unit. The stopper 632 is associated with the shaft portion 632S as a center, but the coil spring, the plate, etc., which changes the state shown in FIG. 3A to the steady state and returns to the steady state after being deformed as shown in FIG. 3C. It has elastic members such as springs. However, the elastic member may not be required as long as the structure naturally returns to the steady state by gravity or the like.

At least a portion 632a of the stopper 632 is exposed outside the case of the power supply unit 600, but is covered with a stopper cover 633 so that the exposed portion cannot be easily pushed or operated with a finger.

With such a configuration, unless the stopper 632 is moved, the safety pin 630 and the stopper 632 collide, and thus the power contact 608 of the power supply unit and the electrode terminal pin 602 of the electrophoresis tank cannot be connected. That is, in the state shown in FIG. 3B, the stopper 632 collides with the safety pin 630 and prevents the electrode terminal pin 602 from entering the inside because it prevents the inside from entering.

Therefore, in this state, it is physically impossible to output power from the power supply unit to the outside, so safety is absolutely ensured.
The safety pin 630 and the electrode terminal pin 602 may be shared. That is, as long as the position of the stopper 632 is not changed, it is sufficient that the power contact 608 and the electrode terminal pin 602 cannot be physically connected.
For this configuration, in order to enable energization while ensuring safety, it is assumed that an electrophoresis tank lid 634 is installed to prevent electric shock, and the electrophoresis tank lid 634 rotates the stopper 632. Thus, the stopper 632 is moved for the first time, and the power contact 608 and the electrode terminal pin 602 can be connected.

  Specifically, a stopper release protrusion 635 is formed on the migration tank lid 634 as shown in FIG. The stopper release protrusion 635 has a shape that enters the inside of the stopper cover 633. When the power supply unit is pushed to the migration tank side from this state, the stopper release protrusion 635 of the migration tank lid 634 pushes the exposed portion 632a of the stopper 632 as shown in FIG. Then, the collision state between the safety pin 630 and the stopper 632 is released. Therefore, the power supply unit can be entered as it is, and the power contact 608 and the electrode terminal pin 602 are connected. With such a configuration, the power supply unit cannot be connected to the electrophoresis tank unless the electrophoresis tank lid is installed, and an electric shock accident or ignition accident caused by mistakenly inserting a finger or a foreign object into the energized electrophoresis tank. Can be prevented. In the system of FIG. 3, the safety mechanism is not functioned due to a failure of the switch or the danger that the buffer solution etc. touches the switch and the circuit is easily short-circuited is avoided. However, in this system, the power supply unit can be connected only after the electrophoresis tank lid is installed in the electrophoresis tank, but the electrode terminal pin 602 and the power contact 608 are fitted with the strength suitable for the power consumption of several tens of watts. Therefore, vibration may be transmitted to the electrophoresis tank at the time of connection, and the installed sample may be affected.

In order to avoid such problems, first, the electrophoresis tank and the power supply unit are connected, then the gel and sample for electrophoresis are installed, and the electrophoresis tank lid is gently installed to enable migration. A procedure is desirable.
Therefore, FIG. 4 shows an example of a system that realizes this procedure and can avoid the problem of the switch failure and the short circuit easily. The migration tank lid 640 is provided with a protruding structure 641. The migration tank lid 640 is for covering the migration tank 642, but it is assumed that the migration tank 642 and the power supply unit 643 are connected before the migration tank lid 640 is installed. As a connection method, for example, electrode terminal pins and power contacts may be used as shown in FIG.
However, power must not be supplied from the power supply unit to the migration tank without the migration tank lid 640 being installed.
In other words, the circuit board 644 incorporated in the power supply unit 643 must be disconnected as an electric circuit when the migration tank lid 640 is not installed, regardless of the status of the migration tank lid 640 switch.

  For example, if two or more relay parts 645 are installed on the circuit board 644 and the parts are not electrically connected, power is not supplied to the migration tank side. Further, a lever 646 is prepared, and a conductive member 647 is attached to the tip of the lever 646, so that the lever 646 can be rotated about the rotation shaft 648. The rotating shaft 648 is more convenient for manufacturing if it is installed on the circuit board 644 as shown in FIG. 4A, but the position is not particularly a problem. What is important is that the conductive member 647 and the relay component 645 are usually separated from each other by a load of the conductive member 647, etc., but if the lever 646 is rotated by applying an external force by the migration tank lid 640, the conductive member 647 is electrically conductive. The member 647 and the relay component 645 are in contact with each other so that the plurality of relay components 645 are electrically connected and energized.

For example, as shown in FIG. 4A, the projection structure 641 of the electrophoresis tank lid is inserted into the opening 649 of the power supply unit, and the lever 646 is rotated to bring the conductive member 647 into contact with the relay component 645. (The contact state is shown in FIG. 4 (b)). In this method, since no switch is used, the distance between the conductive member 647 and the relay component 645 in a normal state can be secured at a sufficiently safe arbitrary size, and the switch is installed on a substrate like a switch. Therefore, there is an advantage that a position where the buffer solution is difficult to be applied and a position where the circuit is not short-circuited easily can be selected and installed. Since the safety is the first, it is natural that the opening 649 has a size (for example, less than 3 mm in diameter) that does not easily allow a foreign object such as a finger to enter.
As described above, according to the present embodiment, at least the structure in which electrical connection is not completed unless migration is possible can be made possible while maintaining the durability of components.

Furthermore, the apparatus and auxiliary instrument regarding the observation after electrophoresis which are one Example of this invention are demonstrated. Many biomolecules such as nucleic acids and proteins that are generally handled as samples for electrophoresis are transparent to visible light. Therefore, fluorescent substances that bind to these biomolecules are used for detection after electrophoresis. Often to do. The fluorescent material used generally absorbs ultraviolet light and emits visible light fluorescence. Ultraviolet light is not only harmful to humans, but also affects film and CCD elements when recording with a camera. Therefore, as an apparatus and an auxiliary instrument relating to observation after electrophoresis, an apparatus for irradiating ultraviolet light and an optical signal obtained from the sample are recorded, and the irradiation light does not affect observation and photographing. Each of the observation and photographing devices to be cut into two is necessary.
The inventors have proposed in Japanese Patent Application No. 2004-73549 and Japanese Patent Application No. 2004-148223 regarding an ultraviolet light irradiation apparatus and an observation imaging apparatus, and further propose an apparatus with improved irradiation uniformity and operability.

First, an ultraviolet light irradiation apparatus will be described. FIG. 5 shows the inside of the ultraviolet light irradiation apparatus according to the present invention. The housing 1600 of the irradiation device is preferably made of a metal such as aluminum, steel, and stainless steel in terms of durability against ultraviolet light, considering chemical resistance, internal ultraviolet light reflection efficiency, light weight, and the like. For example, it is better to use aluminum.
A light source 1601 having a maximum intensity wavelength in the range of 250 nm to 325 nm is disposed inside the housing 1600, and an ultraviolet fluorescent tube is usually used. As a light source that can emit ultraviolet light with a wavelength of 320 nm or less, which is necessary for detection of biomolecules, fluorescent tubes (fluorescent lamps) that use mercury are currently the best. This is because there is a problem that there is no semiconductor light source having a suitable wavelength band. Therefore, if a fluorescent label for biomolecules having an absorption band other than ultraviolet light is used, or if a light source made of a semiconductor or an organic compound having an optimum wavelength can be used, a fluorescent tube is not necessarily used. . For example, recently, a new labeling substance using the quantum effect has been developed.

  The light emission phenomenon using the quantum effect generates different quantum states (energy states) depending on the physical structure such as the particle size of the material itself and the size difference, not the physical properties of the atoms and atomic bonds that make up the material. Even when excited with the same light source using, various emission wavelengths can be realized. A substance in which a substance (molecule) that easily binds to a biomolecule is bound to such a quantum structure has been put to practical use as a label for the biomolecule. Currently, antibodies and antigens are used as binding substances, and they are often used as fluorescent labels for detecting specific molecules. . For example, if a mixture of quantum structures with different particle sizes is introduced into a centrifuge together with a biological sample, the quantum structures having the same mass as each component contained in the biological sample will be distributed at the same position. If each biological component and the quantum structure can be combined by heating or the like in a state, simple mass spectrometry can be performed by analyzing the wavelength distribution of the combined quantum structure. Aside from specific examples, since a new labeling substance has been developed in this way, the light source is not necessarily specialized for ultraviolet light. When the area to be irradiated with the light source covers a wide area, a plurality of light sources 1601 may be arranged as shown in FIG. 5. For example, when irradiating an area of about 150 mm × 150 mm, about six are arranged. Is appropriately selected depending on the size of the gel to be illuminated and the signal intensity of the sample.

If the light sources 1601 are simply arranged at equal intervals, the intensity of illumination is generated at equal intervals, and the non-uniformity of illuminance increases as shown in FIG. FIG. 6A is a distribution diagram in which the vertical axis is the illuminance and the horizontal axis X is the light source arrangement site. Reference numeral 1601 denotes the position of the light source in the distribution diagram of FIG. The arrangement of the light sources 1601 shown in (a), (b), and (c) of FIG. 6 corresponds to a cross section cut along xx ′ with respect to the configuration shown in FIG.
The illuminance shown in FIGS. 6 (a) and 6 (b) indicates a part separated from the light source by about 10 mm to 25 mm (about 1 to 4 times the diameter of the light source).
Assuming that the fluorescent tube 1601 is a linear light source, the light emitted from the partial element Δl of the fluorescent tube is distributed to an area of 2πr · Δl at a distance r, and the illuminance is proportional to 1 / r. The illuminance distribution L on the irradiation surface (horizontal axis) X in FIG. 6 (b) is not simply proportional to 1 / r because the light amounts from a plurality of adjacent light sources are added, but from the fluorescent tube. The maximum value is at a right above the fluorescent tube 1601 where the optical path is the shortest (r1), and the minimum value is at the intermediate point b where the optical path is the longest (r2). If the distance between the light source and the irradiation surface is increased, the difference between the maximum value and the minimum value is reduced. However, the absolute value of the light amount is greatly reduced, and the irradiation device is also increased in size, which is not a practical measure.

  Therefore, in order to improve the uniformity of illuminance, a light reflector 1602 is installed on the back side of the light source 1601 (the direction opposite to the direction in which the sample to be irradiated is installed) as shown in FIG. It is necessary to supplement the amount of light at b with reflected light. Specifically, a configuration as shown in FIG. Of course, the irradiating device has a bottom surface 1600a. If the bottom surface 1600a is formed of a material having excellent ultraviolet light reflection characteristics such as aluminum, the reflected light from the bottom surface 1600a also greatly affects the illuminance distribution L. First, the reflected light path r3 that is reflected by the bottom surface 1600a and enters the vicinity of the intermediate point b of the irradiation surface X should be used as it is. On the other hand, the reflected light path that is reflected by the bottom surface 1600a and enters the vicinity of a immediately above the adjacent light source originally enters a portion where the illuminance by direct light is large, so the non-uniformity of the illuminance distribution L is expanded. I will let you. Therefore, a reflector 1602 is installed to change the optical path that does not enter the intermediate point b if there is nothing into the optical path r4 that enters the vicinity of the intermediate point b. Thereby, the illuminance at the intermediate point b is increased, the difference between the maximum value and the minimum value of the illuminance distribution L is reduced, and the uniformity is improved. In the case where there is no reflector 1602 and only the bottom surface 1600a exists, the nonuniformity is amplified by the reflected light directly above the light source, so that the reflector 1602 not only increases the amount of light at the intermediate point b,

  It also fulfills the function of preventing an increase in the amount of light just above the light source a. The specific shape of the reflector 1602 may be a shape obtained by bending a flat plate as shown in FIG. 6B or a curved surface. What is important is that the optical path r3 that is reflected by the bottom surface 1600a and does not enter the vicinity of the intermediate point b where the illuminance is low does not affect the optical path that does not enter the vicinity of the intermediate point b. The shape should be changed to r4. For example, if the light source interval is p, 60% or more of the reflected light component (the component that does not have a vector from the light source to the irradiation surface direction) is collected near the midpoint b (bp / 3 to b + p / 3). The amount of light that is 1.5 times or more the amount of light that the reflected light component reaches or directly above the light source (ap / 2 to a + p / 2) around the intermediate point b (bp / 2 to b + p / 2) However, the shape is not limited to this. Since it does not affect the optical path r3, it is usually a discontinuous shape as shown in FIG. 6B. However, when the reflection at the bottom surface 1600a is small, the reflection at the bottom surface 1600a is compensated. Further, a shape 1602a as shown in FIG. 6C in which a part of the bottom surface is fused may be used. In terms of non-uniform illuminance distribution, there is only one adjacent fluorescent tube in the vicinity of the outermost fluorescent tube, so the amount of light is insufficient. Therefore, the amount of light on the outside may be supplemented by a method such as narrowing the interval between the outer fluorescent tubes than the interval between the fluorescent tubes at the center.

The light source system composed of the light source 1601 and the light reflection plate 1602 configured as described above is housed in the inner case 1603 as shown in FIG. If the light source 1601 is an ultraviolet light source, the inner case is preferably made of aluminum, and if it is a light source of another wavelength, a material having a high light reflectance for that wavelength is selected. The inner case 1603 concentrates the ultraviolet light on the irradiation surface and prevents the surrounding wiring and electronic components from being deteriorated by being irradiated with the ultraviolet light. However, in this state, the heat generated in the fluorescent tube remains in the case and is difficult to cool.
Therefore, a cross-sectional configuration as shown in FIG. 7 is considered. The inner case 1603 shown in FIG. 5 is easy to manufacture to form a box shape with the upper part opened by overlapping two U-shaped members so as to be perpendicular to each other as shown in the figure. As shown in FIG. 7, the bottom surfaces of these two U-shaped members 1612 and 1613 are shifted in the vertical direction so that a space K is generated. In addition, if a ventilation hole 1614 is provided below the bottom surface or side surface of the lower member 1613, air permeability is secured while maintaining the light shielding function, and cooling can be achieved if internal air is sucked from the ventilation hole 1614 by a fan or the like. Increases efficiency. The upper member 1612 may have a reflective structure as described with reference to FIGS. 6B and 6C.

The light source system configured in the inner case 1603 as described above functions by supplying power from the power supply substrate 1605 to the electrode 1604 of the fluorescent tube in FIG. Usually, one inverter board for driving the fluorescent tubes is often used for one fluorescent tube. Therefore, the power supply board 1605 is not the inverter board itself but distributes electric power from the outside to these inverter boards. Functions as a relay board. An external power cord 1606 is connected to the power board 1605 or a connector component with the external power cord 1606 is installed.
The external power cord 1606 is connected to an outlet as it is if the power required by the irradiation device is commercial AC power, but is connected to an AC / DC conversion adapter if DC power is required. Regardless of which method is used, a high voltage is required for lighting the fluorescent tube, and it is necessary to boost the voltage. Therefore, it is safer to use an AC / DC conversion adapter for external power supply. Convenient in terms of overseas use.

In practice, when the switch 1609 for turning on and off the light source, the light emitting diode 1610 for displaying the lighting state of the light source, and the safety cover for cutting ultraviolet light are opened, the power supply to the light source is forcibly stopped. Electronic parts such as a safety switch 1611 are also required, and wiring with these parts and connector parts thereof are also installed on the power supply board 1605. The safety switch 1611 is, for example, a magnet switch, and a part for connecting the switch to a safety cover side that cuts ultraviolet light is installed.
However, in a configuration in which the light source cannot be driven unless the safety cover is closed, there may be a problem in practical use. Therefore, a release switch that disables the safety switch 1611 by short-circuiting is also installed as necessary. Since the release switch should not be in a state where it can be easily touched, it is desirable that the release switch be disposed at a position where it cannot be operated unless it is intentionally pressed from a small hole formed in the housing 1600 with a thin pin or the like. Reference numeral 1607 denotes a fan, which keeps the internal temperature low by sucking outside air from the fan and removing internal air from the vent hole 1608. In addition, as shown in FIG. 7, a ventilation hole 1614 may be provided on the bottom surface of the housing, or an electric cooling fan may be disposed in the ventilation hole 1614. In any case, it should not be configured such that ultraviolet light leaks from the inner case 1603 shown in FIG. 5 to the outside through the vent hole 1608, and a double bottom as shown in FIG. A device like a structure is necessary.

FIG. 8 shows an example of an ultraviolet light irradiation apparatus. FIG. 8 is preferably used in a dark room. An upper cover 1621 is installed on the internal unit of FIG. An ultraviolet light transmission filter 1622 that transmits ultraviolet light and cuts visible light is disposed at the center of the upper cover 1621. Specifically, this filter transmits 50% or more of wavelengths of 290 nm to 320 nm and cuts 99% or more of wavelengths of 400 nm or more, but the characteristics can be changed depending on the light source and application. The size of the filter is about 60 mm × 110 mm or more and 150 mm × 150 mm or less, but this can also vary depending on the application. Further, it is preferable to connect an ultraviolet light cut cover 1623 to the upper cover 1621 or the casing of the irradiation apparatus for safety. As a connection method, a hinge 1624 or the like can be opened and closed in the vertical and horizontal directions as necessary. It is desirable that the hinge 1624 can be stopped at an arbitrary position, such as a torque hinge (for example, PC-30, handled by Morrigin Co., Ltd.). In this case, the ultraviolet light cut cover 1623 is fixed in front of the face for safety. In the meantime, it is possible to use both hands freely. Reference numeral 1626 denotes a safety switch operating device for operating a safety switch 1625 installed on the ultraviolet light irradiation apparatus side. For example, if the safety switch 1625 is a magnet switch, the safety switch operating device 1626 is a magnet. If the safety switch 1625 is pushed by the weight of the ultraviolet light cut cover 1623, the safety switch operating device 1626 may be unnecessary.
In use, the ultraviolet light cut cover 1623 shown in FIG. 8 is opened, and after the electrophoresis tank capable of transmitting ultraviolet light or the gel after electrophoresis, the ultraviolet light transmission tray and the electrophoresis gel are placed on the ultraviolet light transmission filter 1622, again. It is preferable to measure with the ultraviolet light cut cover 1623 closed.

In the configuration of the ultraviolet light irradiation apparatus as described so far, the fluorescence from the sample is not so strong, so that it can basically be used only in a dark room. In order to perform observation and photographing of a sample even in a bright room, it is necessary to install a simple dark room cover above the ultraviolet light irradiation device. FIG. 9 shows an example of a simple darkroom cover. The simple darkroom cover casing 1701 is formed of a metal such as stainless steel or aluminum from the viewpoint of ultraviolet resistance and chemical resistance. Reference numeral 1702 denotes a safety switch operating device similar to 1626 shown in FIG. 8 for operating a safety switch installed on the ultraviolet irradiation apparatus side. The lower surface of the housing 1701 has a large opening 1703, and an irradiation device is installed below the housing 1701 so that irradiation light is taken into the housing. However, if external light enters the inside of the housing, the observation photographing is hindered. Therefore, light shielding frames 1704 are formed on the four sides around the opening 1703. When it is assumed to be used in an uneven surface, it is difficult to completely shield the light only by the light shielding frame 1704, and therefore, an elastic material such as sponge or rubber is attached to the back surface of the light shielding frame 1704. Furthermore, the left and right sides of the housing 1701 are also open, but a frame 1705 is similarly formed on the four sides around this side.

  The frame 1705 is not a main object of light shielding, but a structure for strengthening the structure and attaching a light shielding curtain 1707. Therefore, members necessary for attaching the light shielding curtain such as a magic tape and a magnet sheet are bonded to the frame 1705. In the simple darkroom cover of the present invention, it is assumed that the user puts his hand inside the housing and operates, so that the wrist 1 will be painful if the frame 1705 remains as it is. Therefore, a cushion member 1706 made of rubber, sponge, or the like is installed at the lower part of the frame 1705 (the part that is touched when the hand is put inside the housing 1701) so that it can be operated smoothly. As described above, reference numeral 1707 denotes a light-shielding curtain, which is attached to the left and right sides of the housing 1701 with a magic tape or a magnet sheet bonded to the frame 1705. The light-shielding curtain 1707 may be removed when a sample or instrument is taken in or out of the housing. Further, when the operation is performed with the hand inside the casing, external light enters when the light shielding curtain 1707 is peeled off. Therefore, it is desirable that the operation can be performed with the light shielding curtain 1707 attached to the frame 1705. For this reason, sleeves 1708 are sewn on the light-shielding curtain 1707, and a hand can be inserted from here. Rubber is put in the exit of the sleeve so that it is in close contact with the wrist and does not allow external light to enter. When the hand is not inserted, external light enters from the sleeve 1708, and thus an outer curtain 1709 is further sewn outside the light-shielding curtain 1707.

The sewing part is preferably the upper part because it is necessary to put a hand inside the outer curtain 1709. If necessary, the outer curtain 1709 is fixed by the lower connecting member 1710 so that light can be completely shielded. The connecting member 1710 is preferably a hook, button, magic tape, or the like that can be attached and detached with one touch. In this way, by forming a side surface with a flexible material having flexibility, and further providing a flexible cylindrical (sleeve-shaped) operation port on the side surface, a very high degree of freedom and complete light shielding Internal processing can be performed while maintaining the characteristics.
When the light-shielding curtain 1707 is attached to the left and right surfaces of the housing 1701, the inside cannot be observed because it is completely sealed. Therefore, an opening 1711 is provided in the upper portion of the housing 1701. Since ultraviolet light is used, a simple opening is harmful to the eyes. Therefore, a transparent member that cuts ultraviolet light is provided inside the opening 1711. In order to operate by putting a hand while observing from above, it is preferable that the opening 1711 is as large as possible, and it should be a hole having a diameter of 80 mm or more. In addition, it is desirable to be able to work while sitting on a chair when performing internal operations, and the height of the casing 1701 is about 140 to 300 mm, but it depends on the height of the ultraviolet light irradiation device installed below. However, it is not limited to this range.

  Since the lower the housing 1701 is, the narrower the range that can be observed and photographed, the irradiation device shown in FIG. 8 employs a small light source such as a cold cathode tube or a light-emitting diode to suppress the height of the irradiation device. Therefore, it is preferable to secure the height of the observation photographing apparatus. A lens barrel 1712 is attached to the upper part of the opening 1711. As shown in the drawing, the lens barrel 1712 covers the periphery of the opening 1711 and prevents outside light from entering the inside of the housing from the lateral direction and the oblique direction. Further, the hole conversion adapter 1713 is installed in a form that covers the upper portion of the lens barrel 1712. A lens opening 1715 having a size of about 37 to 43 mm, which is an optimum size for a camera lens, is provided at the center. The hole conversion adapter 1713 is screwed using a screw hole 1714 provided on the side of the lens barrel 1712. The lens opening 1715 and the camera adapter connection opening 1716 are threaded so as to engage with each other and are connected. A camera fixing screw 1717 of the camera adapter is adapted to fit a tripod fixing screw hole provided on the bottom surface of the camera, and can fix the camera.

  The position of the camera fixing screw 1717 can be freely moved left and right. In addition, since the member to which the camera is fixed can be moved back and forth and up and down by the back and forth moving screw 1718 and the up and down moving screw 1719, the position of the camera can be changed three-dimensionally, and many cameras can be fixed. It is like that. As a camera to be connected, a film camera such as a polaroid camera often used for life science research, a widely used digital camera, a video camera, and the like are assumed. Polaroid cameras are suitable for experiments because they can shoot samples and develop them on the spot, but digital cameras can also be connected to computers and printers via small removable media such as USB memory or connected to small printers. Since data can be transferred, equivalent usage is possible. If the use of cables and media is troublesome, it is also possible to use a camera, mobile phone, mobile terminal, etc. that can transfer data by wireless communication. In any case, since it is common for many users to share experimental equipment, can they use personally owned cameras and mobile devices as a means of photography, or can they output data without having to carry the camera? Desirably, the camera itself is provided with developing or printing means. It is desirable to be able to check on the spot whether the camera is properly focused and whether exposure and color balance settings are appropriate, so it can be enlarged by connecting to a display screen attached to the camera or a camera. It is possible to use a display device that can display.

Various optical filters are commercially available for photographing biological samples, and these filters can be inserted into the upper portion of the connection opening 1716 of the camera adapter as necessary. Further, since it is troublesome to attach and detach the hole conversion adapter 1713 every time of observation and photographing, a simple window 1720 is formed so that the inside of the housing 1701 can be easily observed with the camera connected. Since it is for internal confirmation, the simple window 1720 does not need to be so large, and is about 40 mm × 90 mm. However, since ultraviolet light cannot be viewed directly, a transparent member that cuts ultraviolet light is also installed here. The ultraviolet light cut member of the opening 1711 and the simple window 1720 may be shared. When photographing, outside light should not enter from the simple window 1720, and thus a small cover 1721 that can cover the simple window 1720 with one touch is installed. The small cover 1721 can be opened and closed in the left-right or up-down direction, and is formed of metal or a curtain. Select a connection method suitable for the material, such as hinges or velcro.

  The simple darkroom cover is not necessarily applicable only to the ultraviolet irradiation apparatus of the present invention, and can be used by being placed above a commercially available irradiation apparatus. If it is installed and used in an irradiation device that does not indicate whether ultraviolet light is irradiated by a light emitting diode or the like, it is desirable that the presence or absence of irradiation can be confirmed by some method. Therefore, a fluorescent display member 1722 is installed on the upper surface or the front surface of the housing 1701. This is formed of fluorescent acrylic that emits light with respect to ultraviolet light, and as long as ultraviolet light is irradiated, it emits light regardless of the irradiation device, so the irradiation state can be easily confirmed.

  FIG. 10 is a cross-sectional view of the center portion of the simple darkroom cover of the present invention (a view with the right side of FIG. 9 as the front). A frame structure 1802 formed by folding or welding exists in the grounding portion of the darkroom cover housing 1801 so that external light does not enter the housing. An ultraviolet cut plate 1803 is attached to the back surface of the top opening by screwing or bonding, so that internal observation with the naked eye is possible. A lens barrel 1804 is attached to the peripheral surface of the upper surface opening, and a hole conversion adapter 1805 is fitted on the upper portion thereof. As shown in the figure, an inner wall is formed on the back surface (the lens barrel side) of the hole conversion adapter 1805, and is fixed to the lens barrel 1804 with a fixing screw 1806 as necessary. In order to ensure fixation by the fixing screw 1806, a groove may be provided in a portion where the hole conversion adapter 1805 contacts the tip of the fixing screw 1806. A stand support 1807 is provided on the rear surface of the housing 1801 and a stand 1808 is attached thereto. As shown in FIG. 10, when it is unnecessary, the stand 1808 is in contact with the back surface of the housing 1801, and if necessary, the base is rotated and fixed at a specified angle, and the stand 1808 is placed so that the tip of the stand 1808 is in contact with the installation surface. Thus, the entire simple darkroom cover can be fixed open.

  When a large sample or instrument is taken in or out, it is more convenient to open and close the whole as described above. A lens support 1809 is provided on the inner surface of the housing 1801, and a lens frame 1810 fitted with a lens 1811 is attached to the case 1801. The lens frame 1810 can be rotated in the same manner as the stand 1808 and can be fixed at a specified or arbitrary angle. Although it is more convenient to use a lens that can be magnified when processing fine parts inside the sample, the lens is a hindrance during shooting, so you can easily use the lens as necessary. A structure that can be removed (moved) is desirable. A lighting unit 1812 can be attached to and detached from the top of the housing 1801 as necessary. The light source 1813 incorporated in the illumination unit 1812 is a fluorescent tube, a light emitting diode, an organic light emitting material, a laser, or the like, and the emission color is white for a general colored sample, and single color for a sample that requires specific excitation light. Or ultraviolet light is selected. A reflector 1814 is installed behind the light source 1813 as needed, and has the effect of dispersing light so that the sample can be condensed or irradiated over a wide range. When the light source with strong directivity or the light condensing effect by the reflecting plate is high, the light diffusing plate 1815 provided between the light source 1813 and the irradiation target diffuses light so that uniform irradiation is possible. To do. Electric power is supplied to the light source 1813 from the power cord 1817 via the light source driving board 1816.

A sample can be photographed by the irradiation apparatus and the simple darkroom cover described so far, and image data can be obtained. When a digital camera is used as a photographing machine, image data can be directly taken into a computer as digital data. When a film camera is used, it is a common practice to digitize data using an optical scanner or the like and load it into a computer. In any case, it is necessary to analyze the digital image data captured in the computer by software. Next, the image analysis software of the present invention will be described.
The image analysis software of the present invention (hereinafter referred to as “this software”) is assumed to be used in a widely used personal computer, and its operating system is Windows (manufactured by Microsoft Corporation). It is assumed that the system operates on a system based on window display on the display screen and operations using mouse input and keyboard input, as represented by MacOS (manufactured by Apple Computer). FIG. 11 is an overview of the operation of this software, and adopts a general multi-window format. That is, the main window 1901 that controls the whole has a window name display portion 1902 and a command menu portion 1903 at the top, and each subwindow 1904 and palette window 1906 are displayed and operated inside the main window. The sub window 1904 is for performing image processing and analysis processing, and displays an object 1905 such as an image or a graph as necessary. Details will be described later. The palette window 1906 supplements the command menu unit 1903, and frequently used commands (commands) are preferentially displayed with characters and buttons, or set by the user. This will also be described later.

First, an image processing window among the sub-windows will be described. The purpose of this software is to analyze image data obtained by experiments such as electrophoresis, so an image processing window for displaying and processing image data is required. An overview of the image processing window is as shown in FIG. There is a window name display section 2002 at the upper part of the image processing window 2001. If it is not specified, the name of the image file to be displayed is displayed. However, it can be changed by specification. In the lower part, there is a status display unit 2003, which displays the position coordinates of the mouse pointer, image pixel data (luminance, etc.) at that position, the enlargement ratio of the image display, and the like. Reference numeral 2004 denotes an image displayed by opening image data, and the enlargement ratio and the like can be changed. A “graphic object” such as a rectangle, an ellipse, a polygon, a straight line, a broken line, or the like can be drawn on the image 2004 with a mouse. The type of figure is specified from the command menu of the main window, the right-click menu of the image, the command button on the palette window, and the like. The generation timing of the graphic may be either a method of drawing on the image immediately after specifying the graphic and transforming it with the mouse, or a method of specifying the position and size with the mouse after specifying the graphic.

  With the former method, there is room to devise such as generating the same type of figure according to the previous setting, but it is still difficult to realize the shape desired by the user from the beginning, so the drawn figure is The operation to deform is a prerequisite. For example, in the transformation of a figure, the whole of the rectangle is circumscribed by dragging the corner of the rectangle that circumscribes the figure, or each vertex of the figure is moved individually by dragging. A figure can also be generated by copying and pasting exactly the same thing. For images and graphic objects, properties (characteristic parameters) can be changed by inputting from a keyboard into a “property window” displayed by clicking or double-clicking with a mouse. Changes from the property window are the name, display color, size, position, attribute, hierarchical structure, and display switching of the image and graphic objects (which figure is displayed if it is an image, whether it is displayed if it is a figure) Or the like. These will be described later. In addition, since the shape deformation method described above cannot increase the number of vertices of the shape, this property window has a button to enter the mode for adding or deleting vertices. If you specify with the mouse, the vertex is added or deleted at that position.

In order to analyze image data such as electrophoresis, first, necessary image processing must be performed on the obtained image data. The image processing functions possessed by this software include the following.

Monochrome When the image data is a color image, the analysis is often cumbersome as it is, for example, what color is selected as a luminance reference or graphing target. The color to be analyzed in the experiment is usually narrowed down such as the color of the reagent used. Therefore, it is desirable to make the image monochrome (single color) and simplify the subsequent analysis processing. Monochrome processing performed by this software is a method that extracts only a specific color from a color image, and a method that calculates an average value (including mean square, factorial average, etc.) by multiplying each color component by a user-specified coefficient. Etc. A two-gradation process is also included in which a value above a certain reference value is converted into a maximum value, and a value below the reference value is converted into a minimum value.

Color correction This is a process of performing normalization after multiplying each color component by a user-specified coefficient and converting it into a color component after conversion.

This is a process of applying a conversion function having the current luminance value as a variable to each color component of contrast correction and converting it to a color component after conversion. The conversion function is not limited to that expressed by a mathematical expression, but can be specified by deforming a graph indicating the relationship between the luminance value before conversion and the luminance value after conversion with a mouse or drawing a broken line.

This is a process for changing the size of an image to be displayed.

The process of rotating the entire image or rotating the image in the graphic object. A method to display the property window of the image or graphic object and specify the rotation angle numerically, a method to determine the rotation angle from the mouse displacement by dragging with the mouse, a method based on the graphic object drawn on the image is there. In particular, in the case where the sample has directionality, such as an electrophoresis image, for example, a straight line (or a broken line) connecting the starting points of electrophoresis is used as a reference so that it becomes a desired angle such as horizontal or vertical. The method of rotating the entire image is easier to use than other methods. As a procedure, a graphic object such as a straight line is drawn on a portion desired to have a desired angle on the image, and a command “Rotate” − “Rotate based on a specified graphic” is executed. When this command is executed, a vertical bisector is temporarily displayed for the figure to be rotated, and by clicking a point on it, the rotation center and the rotation axis (vertical bisector) are changed. It is specified. When a desired angle such as “horizontal”, “vertical”, or “45 °” is selected from the dialog to be opened next, the image is rotated. If the image is not a square or a perfect circle, the image size before and after rotation is different, so the image size is enlarged so that all the figures before and after the rotation are included, and there is no luminance value data Data such as white and black will be supplemented.
This is a process of inverting the brightness value of the brightness value inverted image so that the maximum value and the minimum value are switched. That is, the maximum luminance value handled by the software is X, the luminance value of each pixel is x, and x is converted to X−x. For color images, the same processing is applied to each color component such as (R, G, B).

After appropriately executing the image processing command, the process proceeds to analysis processing. The analysis may be performed on the entire image, but when the area where the sample exists is limited or regular as in electrophoresis, it is common to perform analysis on a specific area. is there. In this software, the area is specified by drawing the various graphic objects already described. Even if the figure is the same, there are various types of analysis processing to be performed. Therefore, in this software, appropriate analysis processing is performed by designating or changing the attributes and hierarchical properties of the drawn figure. These designations and changes are executed by double-clicking the graphic object or selecting it from the command menu and entering it in the “Property Window” that opens. The method of selecting attributes from the right-click menu is also convenient, so that it can be used. Hereinafter, attributes and a hierarchical structure will be described.

Lane attribute A rectangular or polygonal object attribute. In the case of electrophoresis using a flat type gel, it is common to run a plurality of samples simultaneously, and the course in which each runs is called a lane. If a rectangle surrounding this lane is drawn like the rectangle in the graphic object 2005 of FIG. 12 and the attribute of the rectangle is designated as “lane”, the region surrounded by this rectangle will be recognized as an electrophoresis lane. It becomes the object of analysis processing for electrophoresis. The same applies to polygonal objects. As will be described later, since this software has a function of automatically recognizing a plurality of lanes, the attribute of the lane automatically recognized (generated) by the software in this way is also “lane” from the beginning.

Marker lane attribute A rectangular or polygonal object attribute. The basic characteristics are the same as the lane attributes, but the general lane contains a band with unknown molecular weight (described below), while the marker lane has a known molecular weight (and substance weight). It is characteristic that it consists of. Since there are a plurality of samples used as markers, each has a unique name. In this software, data related to markers is collected in one file, and if the marker name, the number of included bands, the molecular weight of each band, the amount of substance, etc. are arranged in one line, and multiple markers are registered , This line also spans multiple. It is also possible for the user to register a marker independently, or to additionally register new marker data through the Internet. In these cases, a new line is added to the marker data file.

  When the user designates the rectangular and polygonal objects (including those already specified in the lane attribute) on the image, if the attribute is specified as “marker lane”, the marker is selected. To display the marker data file information in a tabular format. A marker name is displayed in the first column of each row (preferably in alphabetical order), the marker name column is in the form of a button, and the selection is confirmed by pressing the mouse. This dialog is also used when a user registers a marker independently, and it is easy to use if information such as the marker name, the number of bands, and the molecular weight can be directly entered in the displayed table. If a partial image surrounded by marker lanes is also displayed in this dialog, the possibility of wrong selection is reduced.

Band attribute A rectangular or polygonal object attribute. A sample that migrates in each lane by electrophoresis is separated into a plurality of bands 2007 as shown in FIG. 12 mainly in accordance with the molecular size in the course of electrophoresis. This is also basically a rectangle, and if the attribute of the surrounding graphic object is designated as “band”, it becomes a target of analysis processing normally performed on the band such as molecular weight calculation in the analysis processing. Since the band belongs to the lane, the name of the lane corresponding to the upper hierarchy must be specified for the graphic object whose attribute is specified as “band”. However, if the object designated as the band is located inside the object having the specific lane attribute at that time, the lane designation is automatically performed. As will be described later, this software has an automatic band recognition function, and usually performs automatic recognition processing on objects with lane attributes. , The attribute is “band”, and the upper lane is a lane containing the band.

Range attribute This is an attribute of a rectangle, polygon, or ellipse object. This is used when you want to specify any two-dimensional area (range) on the image. For example, it is used when it is desired to calculate the luminance average value or integral value of a specific area on the image. In many cases, it is necessary to perform the same processing for a plurality of ranges. However, it is preferable to determine whether a plurality of graphic objects having range attributes are grouped or their names. For example, if an analysis processing command is selected for one of the objects with a name that differs only in the index in the format XXX [i] (i is an index), the same processing is performed for objects with the same name. You can open a dialog asking if you want to do.

Center attribute line (including broken line) This is an attribute of an object, and indicates the center line of a lane or band. When creating a gray scale graph, it is a common process to graph along a line having a center attribute.

Well line attribute This is an attribute of a line (including broken line) object. In the analysis of electrophoresis, since the moving distance of the band is important, it is necessary to clearly recognize where the starting point of electrophoresis (the well where the sample is placed). When multiple samples are run at the same time, the starting point of each lane may be slightly shifted or the image may be tilted diagonally, creating an object that connects the starting points (wells) of each lane with lines. In addition, it is preferable that the attribute is designated as “well line” and the intersection with each lane (or its center line) is recognized as the starting point. Although the well exists as shown in 2006 in FIG. 12 as an image, it is often reflected at a lower brightness than the background. Therefore, the well is automatically recognized during automatic lane recognition or automatic band recognition. Can also be generated. In this case, if the recognition is not successful, the user is allowed to deform and correct the automatically recognized well line.

Band group attribute line (including broken line) object attributes. Among a plurality of bands present in a plurality of lanes, a band group is formed by connecting the center lines of bands having the same molecular weight with a broken line. In principle, if the molecular weight is the same, the bands are arranged in a straight line. However, in general, due to the temperature distribution in the gel, the difference in electrical conditions, and the like, the band is not generally linear. If the same molecular weight band is not a straight line (not equal migration distances), there will be a problem in the analysis. Therefore, it is necessary to designate the same molecular weight band as necessary. As a designation method, a plurality of bands are selected with a mouse, and “well line generation” is executed from the analysis menu or right-click menu of the main window.

Smiley attribute An attribute of a line (including a broken line) object. As described in the band group attribute, the mobility of bands with the same molecular weight is not equal in each lane (generally called the “smileing phenomenon”), but bands with the same molecular weight are in multiple lanes. In this case, the difference in mobility of each lane cannot be designated as a band group. However, if each lane contains a band with a known molecular weight, it is possible to estimate the difference in mobility. Therefore, drawing a broken line using the mouse and expressing the estimated mobility difference can be used as a reference for molecular weight calculation. Specifically, the polygonal line at this time is designated as a smiley attribute, and it is sufficient to input how much the smiley line corresponds to the molecular weight. In addition, if there is a band with a known molecular weight in each lane even if it is not the same, and the molecular weight is given in a table or the like, a smiley line can be automatically generated.

By the operations so far, the basic processing for the image and the input of basic information necessary for the analysis are completed. Next, the process proceeds to a specific analysis process. Since the analysis processing is mainly performed by opening the analysis window, the analysis window will be described here.
FIG. 13 shows an overview of the analysis window of this software. In the upper part of the analysis window 2100, there is a window name display portion 2101 like other windows. It can be changed, but if not specified, the selected graphic object or table data name is displayed. Since graphing is often performed at the time of analysis, a graph display unit 2102 is arranged at the upper left of the analysis window. The original data of the graph is often a graphic object on the image. When a command such as “Create graph” or “Create analysis window” is executed on the graphic object, the graph display unit 2102 will display the graph from the beginning. Is displayed in a drawn state. The graph in this case uses the luminance of the image as the base data.

  The graph display unit 2102 naturally displays a line 2103 indicating a graph, but in addition to this, a peak line 2104 indicating a peak, a range object 2105 indicating an analysis range, and each point on the graph are highlighted. A graph pointer that can be moved with the left and right keys and the up and down keys on the keyboard is also displayed. A graphic object image 2106 that is the basis of the graph is displayed beside the graph display unit 2102. Thereby, it is possible to easily grasp what the graph means. Of course, when the basic data of the graph is not obtained from the image, the graphic object image 2106 is not displayed either. When a graph is generated from a plurality of graphic objects (for example, graphing for a plurality of electrophoresis lanes, etc.), a plurality of graph lines 2103 are also displayed, and the respective graphic object images 2106 are displayed side by side. .

To generate multiple graph lines, execute commands such as “Create graph” and “Create analysis window” from the state where multiple graphic objects are selected, or execute these commands for one graphic object. This is done by searching for a graphic object having the same attribute and opening a dialog asking whether to graph the searched object together. Also, in the “Graph Property Window” that is opened by double-clicking the graph, describe the object name and check box that have the same attribute side by side, and the user selects the object that he / she wants to display in the graph. , Graph lines to display are added / removed at any time.
A table 2107 is displayed below or beside the graph display unit 2102. First, numerical data on which the graph line 2103 is based is displayed in this table. The first column of the table 2107 is a data name 2108, and for example, the base object name is displayed. The data name 2108 is in the form of a button, and some operations are performed by pressing this button depending on the attribute of the object.

For example, when graphing including an object having the marker lane attribute is performed, a marker name is entered in the data name 2108. By pressing this, a dialog for selecting a marker is opened, and appropriate marker data is displayed. It is also possible to change to. However, if appropriate processing cannot be performed, such as when the number of peak lines 2104 detected or specified in the graph line for the marker lane does not match the number of bands of the selected marker, a message box is displayed that warns the user. In some cases, the process is displayed and nothing is performed. Numeric data and character data are displayed in a cell 2109 arranged beside the data name 2108. Here, analysis processing results such as concentration integration, and the presence or absence of peak lines (when peak detection is performed, a peak line display line is provided in the line below the numerical data in the graph, where the peak is located. ○ is displayed).
In addition, the cell 2109 not only displays previously determined results such as the selected numerical data and the presence or absence of a peak, but is also used as an active input / change means. For example, if you click a blank cell in the peak line display row, a new peak line is generated at that position and displayed on the graph, or conversely, if you click a cell that is a peak line, it becomes a blank cell, There is a function to be deleted. This is a necessary function because the automatically recognized peak line does not always match the user's desired one.

  In addition, processing for generating a range object 2105 on the graph is also performed by providing a range designation row next to the numerical data row of the graph and clicking each of the first and last cells of the range. When the value of a cell that originally contains numerical data is changed, there is also a process of changing the shape of the graph line according to the result. Since this table 2107 is closely linked to the graph display unit 2102, when the graph display unit 2107 is clicked, the display of the table 2107 is displayed so that the numerical data of the clicked point is displayed at a desired position such as the center. change. This feature allows you to click on the peak line on the graph to display the peak line cell in the center of the table, click that cell to delete the peak line, and click another cell to create a new peak line. It is possible to make the processing easier by linking the graph and the table so that they appear. In addition to the contents described so far, calculation lines indicating the result of applying a function such as a formula to a numerical data line, and a character input line that displays a character at a position on the graph corresponding to the cell when a character is input to the cell Exists. It is also possible to read numerical data from an external data file and display it on each line, or to generate a graph based on the data.

An analysis command button 2110 is arranged at an appropriate position such as next to or below the table 2107. Commands such as integrating the luminance value for the selected graph line, table row, and range object, inverting the graph display, automatically detecting the peak of the graph, and calculating the molecular weight of the electrophoresis band are buttons. Is displayed. If there is a problem with the molecular weight calculation, such as when no well line is specified or when no marker is specified, a message window will be displayed explaining that it is not appropriate to execute the command by pressing the command button. Cancels execution of. These commands can also be executed from the command menu of the main window.

As a sub-window of this software, there is a command palette window in addition to an image processing window and an analysis window. This window is a palette of frequently used items that saves you the trouble of selecting from a vast number of commands included in the command menu. There are the following methods for selecting commands included in this command palette, and they may be displayed in parallel on the palette as needed. In either method, since the command to be selected and displayed is different for each user, it is desirable to recognize who the current software user is. For example, a method of selecting a user or inputting a password is used when starting the software.

In this method, the user selects a command included in the selection palette by the user according to his / her own intention. Deleting or adding commands can be done at any time by the user.

Select in order of frequency of use Based on the past software usage results, the most frequently used items are arranged on the command palette with priority. The usage record information can be reset according to the intention of the user.

This is a method of recognizing a command executed immediately before selection in the order of connection usage frequency , and placing it on the command palette preferentially in descending order of frequency of use as a command to be used next to the command. In the method of simply displaying in order of the overall usage frequency, the command that the user wants to use is not necessarily displayed. For example, the molecular weight calculation command is not high in the overall use frequency, but it is presumed that the frequency is overwhelmingly high as a command used next to the marker designation command and the peak detection command. Therefore, it is convenient that the molecular weight calculation command is displayed on the command palette immediately after these commands are executed. In this method, each time a command is executed, the command palette also changes. As statistical data, it is necessary to hold the frequency of the next command to be used for each command. It is desirable that this statistical data can be selected for each user. Further, it is possible to reset or edit the data file as necessary. It is also convenient to have a function that registers a series of commands in advance as a macro and automatically executes them continuously.

  Many commonly used analysis software supports network connections, has the function of taking in the latest data through the Internet and using it for analysis. This software also has a function of utilizing data that can be obtained through the Internet or an intranet, or data included in a server on which the software operates, for analysis. With respect to data included in the server, the user who has input the data can set whether or not to grant the authority to allow other users to use the data. In addition, this software not only shares data between networks, but also performs analysis processing through the network as necessary. For example, the analysis processing is distributed to a plurality of computers (at least hardware having an arithmetic processing function) linked by a network, and each analysis result is collected through the network and provided to the user. Like analysis of electrophoresis results, each analysis process is relatively simple, but in an analysis that requires high-throughput processing (a large number of processes are processed simultaneously), the processing is performed on multiple computers. Dispersion is easy, and high-speed processing can be realized at low cost.

As described above, this software allows objects such as images that are the main result of electrophoresis results, graphs and numerical data obtained from them, to hold parameters that are important in electrophoresis analysis as properties, and handle them flexibly. By providing an interface that can be used, we have realized analysis software that can be used intuitively and with a high degree of freedom.

  As described above, the electrophoretic device of the present invention realizes a structure with excellent durability and safety that prevents failure and accidents, while having a very simple circuit and component structure, and is capable of dynamically generating bubbles and buffer solutions. Stable performance is secured by controlling the characteristics. In addition, the electrophoretic observation photographing apparatus of the present invention is a combination of a light irradiation device whose uniformity is improved by devising a light source arrangement and a light reflecting plate structure, a casing made of metal, and a flexible curtain. It is composed of a dark room type photographing apparatus that realizes light-shielding properties and improved usability, and as a whole, it is possible to observe and record electrophoresis results with very simple and high accuracy. In addition, by providing an interface that allows objects such as windows and figures to have useful properties for electrophoretic analysis and to change these properties flexibly, it is possible to prepare analysis software that can be used intuitively and with a high degree of freedom. In addition, an electrophoresis system that can complete a series of experimental procedures from electrophoresis to observation photography and analysis processing is extremely simple and highly accurate.

  The present invention relates to a power supply integrated apparatus for electrophoresis experiments, a power supply with a timer for supplying an electric output to a plurality of integrated electrophoresis apparatuses, a photographing observation apparatus, analysis software, and the like.

The figure which shows one Example of this invention. The figure which shows one Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention. The figure which shows the other Example of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Electrophoresis tank 2 Buffer 3 Platform-like part 3a Gel installation member 3b Path | route structure part 3c Bubble induction part 4 Electrode 4a Cathode 4b Anode 5 Electrophoresis carrier 6 Sample installation groove 7 Sample 8 Electrophoresis tank lid 9 Bottom partition plate 630 Safety pin 631 Safety pin insertion port 632 Stopper 633 Stopper cover 634 Migration tank lid 635 Stopper release projection 640 Migration tank lid 641 Projection structure 642 Migration tank 643 Power supply unit 644 Circuit board 645 Relay component 646 Lever 647 Conductive member 648 Rotating shaft 649 Opening 1600 Housing Body 1600a Bottom face 1601 Light source 1602 Light reflector 1603 Inner case 1604 Fluorescent tube electrode 1605 Power supply board 1606 External power cord 1607 Fan 1608 Vent 1609 Light source switch 1610 Light emitting diode 1611 Safety switch 1612 U-shaped member 1613 U-shaped Material 1614 Ventilation hole 1621 Upper cover 1622 Ultraviolet light transmission filter 1623 Ultraviolet light cut cover 1624 Hinge 1625 Safety switch 1626 Safety switch operation device 1701 Case 1702 Safety switch operation device 1703 Opening 1704 Light shielding frame 1705 Frame 1706 Cushion member 1707 Light shielding curtain 1708 Sleeve 1709 Outer curtain 1710 Connection member 1711 Opening 1712 Lens barrel 1713 Hole conversion adapter 1714 Screw hole 1715 Lens opening 1716 Connection opening 1717 Fixing screw 1718 Back and forth moving screw 1719 Vertical moving screw 1720 Simple window 1721 Small cover 1722 Fluorescent display Member 1801 Case 1802 Frame structure 1803 Ultraviolet cut plate 1804 Lens barrel 1805 Hole conversion adapter 1806 Fixing screw 1807 Stand support 1808 Stand 1809 Lens support 1810 Lens frame 1811 Lens 1812 Illumination unit 1813 Light source 1814 Reflector 1815 Light diffuser 1816 Light source drive board 1901 Main window 1902 Window name display 1903 Command menu 1904 Sub window 1905 Object 1906 Palette window 2001 Image processing window 2002 Window name display section 2003 Status display section 2004 Image 2005 Graphic object 2006 Well 2007 Band 2100 Analysis window 2101 Window name display section 2102 Graph display section 2103 Graph line 2104 Peak line 2105 Range object 2106 Graphic object image 2107 Table 2108 Data name 2109 Cell

Claims (5)

In an electrophoresis apparatus composed of a buffer solution in which an electrode is disposed on both sides, an electrophoresis gel is disposed between the electrodes, and at least the electrophoresis gel is immersed, at least in the vicinity of the electrode An electrophoresis apparatus comprising a partition plate having an inclination arranged so as to protrude from a buffer solution. The electrophoresis apparatus according to claim 1, further comprising a second partition plate directed upward from the bottom of the electrophoresis tank at a position closer to the electrophoresis gel than the inclined partition plate. In an electrophoretic observation apparatus having a housing formed of a material that blocks external light and having at least one side surface opened and a light shielding cover covering the opened side surface, a mouth portion is provided on the light shielding cover side surface. Electrophoresis observation device. The electrophoresis observation apparatus according to claim 3, wherein the light shielding cover is formed of a flexible material. The electrophoresis observation apparatus according to claim 3, wherein a mouth provided on a side surface of the light shielding cover is formed in a cylindrical shape having flexibility.

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