JP4996235B2 - Trace magnetic drug detection device and magnetic drug detection method - Google Patents

Abstract

A technique of measuring detectable trace magnetic particles with high sensitivity and high efficiency three-dimensionally by a magnetic method using a SQUID magnetic sensor. Disk-shaped sample holders are rotated and inserted from the outside of a magnetic shield into the inside thereof one by one. Magnetizing means magnetizes the standardized specimen outside the magnetic shield. A magnetic sensor detects a magnetic field generated from the magnetized standardized specimen inside the magnetic shield. Thus, the detection of the magnetic field and the magnetization are performed in parallel to each other. The specimen is continuously cut, and the magnetic sensor measures the magnetic field generated from the magnetized standardized specimen before and after the cutting.

Description

  The present invention relates to a trace magnetic drug detection device and a magnetic drug detection method.

  In recent years, treatments for reducing dosage to patients by increasing the efficiency of drug accumulation in cancer affected areas by DDS (Drug Delivery System) have been studied. This treatment is gentle on patients because of reduced side effects, and is promising as a future medical technique for efficiently killing cancerous areas.

  One means for guiding a drug administered into the blood to the affected area is, for example, as shown in FIG. This is a magnetic induction means in which a magnetic drug in which a minute drug 51 is bound directly or indirectly around a magnetic particle 50 having a particle diameter of around 100 nanometers, directly or indirectly, is dispensed into blood and disposed outside the body. The magnetic drug is guided in a predetermined direction at the branch of the arterial blood vessel of the patient. Thereby, it is possible to artificially increase the accumulation efficiency of the magnetic drug in the vicinity of the affected area and guide it. In this DDS, it is indispensable for the development research evaluation of the magnetic induction means and the magnetic drug itself to measure whether the nano-sized magnetic drug particles are quantitatively accumulated in the vicinity of the affected part.

  On the other hand, as a general method of immunoassay, a detection antibody that selectively binds to an antigen to be detected is labeled with an optical marker such as a fluorescent enzyme, and an antigen-antibody binding reaction is detected by an optical signal from the optical marker. And an optical method for detecting the type and amount of the antigen. However, in the optical method, regarding the light source inside the cell, the cell outside the light source inhibits light transmission, and the detection sensitivity is not sufficient. Therefore, quantitative measurement cannot be performed sufficiently, and a process of completely washing away unbound optical markers that reduce detection sensitivity is required.

  In recent years, a magnetic method for quantitatively detecting a minute amount of magnetic fine particles using a SQUID (Superconducting Quantum Interference Device) magnetic sensor has been proposed as a method for obtaining detection sensitivity exceeding that of an optical method. In this magnetic method, a very small amount of magnetic fine particles are detected using a very sensitive SQUID magnetic sensor.

A method for detecting a minute amount of magnetic fine particles will be described below. (1) Measurement of magnetic susceptibility;
(2) A method based on measurement of residual magnetism has been proposed.

Hereinafter, (1) and (2) will be described.
(1) Method for measuring magnetic susceptibility:
A DC magnetic field that magnetizes a small amount of magnetic fine particles is applied from a direction perpendicular to the magnetic flux detection direction of the SQUID magnetic sensor, and the change in the magnetic field generated by the small amount of magnetic fine particles moving in the magnetic flux detection region of the SQUID magnetic sensor is measured. (For example, refer to JP 2001-33455 A).

Further, an alternating magnetic field is applied to a very small amount of magnetic fine particles, and the magnetic fine particles are detected by using a SQUID magnetic sensor (see, for example, Japanese Patent Laid-Open No. 2001-133458).
(2) Method for measuring residual magnetism:
When the size of the magnetic fine particles increases, the residual magnetism of the magnetic fine particles does not relax. A magnetic field of about 0.1 T is applied to a minute amount of magnetic fine particles at a location away from the SQUID magnetic sensor, and residual magnetization is generated in the magnetic fine particles. Thereafter, the substrate on which the sample is placed is moved, and the residual magnetization is measured with a SQUID magnetic sensor (see, for example, Japanese Patent Publication No. 10-513551).

  A specific example of a magnetic method for detecting a minute amount of magnetic fine particles described in JP-A-2004-068645 will be described.

FIG. 2 is a diagram for explaining a conventional magnetic particle inspection apparatus using a minute amount of magnetic particles.
FIG. 2 is a diagram including a cross-section showing a partial configuration of a magnetic particle inspection apparatus according to the prior art, and explains the arrangement state of components during measurement of the apparatus.
The plurality of disk-type sample holders 2 are fixed on the circumference by a non-magnetic disk-type sample holder 3 (hereinafter simply referred to as a sample holder 3). The sample holder 3 is rotated by the rotation mechanism 4. The rotating mechanism 4 is held on a moving stage 5 so as to be movable in a three-dimensional direction. By moving the rotating mechanism 4 on the moving stage 5, the sample holder 3 is moved into the magnetic shield 1 and its position is adjusted.

A high-temperature superconducting SQUID 11 (hereinafter simply referred to as a magnetic signal of a minute amount of magnetic fine particles)
SQUID 11) is cooled to a superconducting transition temperature or lower by liquid nitrogen 7 through sapphire rod 9 and copper rod 10, and is composed of an outer tank 6 a of a vacuum heat insulating container and an inner tank 6 b of a vacuum heat insulating container. 6 is thermally shielded from the outside. The vacuum heat insulating container 6 is made of a nonmagnetic material such as SUS or FRP. Exhaust of vaporized nitrogen and supply of liquid nitrogen are performed through the exhaust / supply port 8 into the inner tank 6b of the vacuum heat insulating container 6.
The SQUID temperature raising light source 13 is used to heat the SQUID 11 and remove the trap magnetic flux.

  The SQUID 11 is surrounded by the magnetic shield 1 in order to reduce the input of environmental magnetic noise. The magnetic shield 1 is made of a high magnetic permeability material such as permalloy, and has a three-layer structure made up of magnetic shields 1a, 1b, and 1c. In order to further improve the efficiency of the magnetic shield, it is desirable that the magnetic shield 1 has a multilayer structure.

  A notch 19 is formed in a part of the magnetic shield 1 (1a, 1b, 1c). At the time of measurement, a part of the disk-type sample holder 2 and the sample holder 3 is completely replaced when the disk-type sample holder 2 is replaced from the notch 19 or when the control circuit of the SQUID 11 is adjusted. The magnetic shield 1 is exposed to the outside.

  With this configuration, the influence of the magnetic field generated by the permanent magnet or the magnetic field application coil on the SQUID 11 is reduced. Therefore, while measuring the magnetic signal of a minute amount of magnetic fine particles in the disk type sample holder 2 inside the magnetic shield 1 with the SQUID 11, the minute amount of magnetic fine particles in another disk type sample holder 2 outside the magnetic shield 1 is measured. By applying a magnetic field to the sample holder 3 and rotating the sample holder 3, the magnetic field generated from the minute amount of magnetic fine particles in the disk-shaped sample holder 2 inside the magnetic shield 1 is measured, and other magnetic elements outside the magnetic shield 1 are measured. Magnetization of a minute amount of magnetic fine particles in the disk-shaped sample holder 2 is performed simultaneously or in parallel.

JP 2001-33455 A JP 2001-133458 A Japanese National Patent Publication No. 10-513551 Japanese Patent Laid-Open No. 2004-068645

  In a cell tissue containing cancer cells (hereinafter referred to as a test cell tissue), for example, how much a minute amount of magnetic fine particles delivered in a state where a drug component is suspended by means such as magnetic induction is accumulated. Therefore, in order to put into practical use a minute magnetic particle measuring and testing apparatus that can measure the distribution in two dimensions and three dimensions by a magnetic method, efficient measurement of a sample of a test cell tissue is required. The The prior art magnetic method focuses on how to measure a minute amount of magnetic fine particles, and no concrete proposal has been made for a method for measuring a two-dimensional and three-dimensional minute concentration distribution. .

  In the conventional magnetic method, a method of measuring one or a plurality of samples on a disk container containing a cell tissue sample or a minute amount of magnetic fine particle sample is employed, and individual measurement of each sample is performed. It was only.

  In the above-described prior art, a microscopic observation image is obtained while thinly slicing the slice surface of the matrix end surface of the examination cell tissue with an optical microscope, and each tomographic cross-sectional image of the matrix of the examination cell tissue is visually reconstructed. Although electronic multilayer image forming means for visualizing three-dimensional and three-dimensional images is provided, it is not intended to measure a minute amount of magnetic particles.

  An object of the present invention is to use a magnetic fine particle and a SQUID magnetic sensor to detect a minute amount of magnetic fine particles by a magnetic method and to detect a two-dimensional and three-dimensional minute concentration distribution efficiently with high sensitivity. To provide measurement technology.

The object is to provide a sample holder for holding a sample holder for storing a specimen of a cell tissue containing a trace amount of a magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disc type sample holder. Rotating means for rotating around the central axis, elevating means for raising and lowering the specimen on the disk type sample holder, cutting means for cutting and removing a part of the specimen, and removing a part of the specimen A plurality of image acquisition means for magnifying a portion; magnetizing means for magnetizing the specimen; a magnetic sensor for detecting a magnetic field generated from the magnetized labeled specimen; and a magnetic shield surrounding the magnetic sensor; the disk-shaped sample holder is configured to be inserted from the outside to the inside of sequentially the magnetic shield by the rotation of the disk-shaped sample holder, wherein the magnetizing means the labeled analyte Magnetized in serial magnetic shield outside, the magnetic sensor detects a magnetic field generated from the labeled analyte which is magnetized in the interior of the magnetic shield, and detecting said magnetization of said magnetic field is performed in parallel, the The specimen is continuously cut and configured to measure the magnetic field generated from the labeled specimen magnetized by the magnetic sensor before and after cutting, and measured before and after the cutting process when detecting the magnetic field. This is achieved by a trace magnetic drug detecting device characterized in that the difference is calculated from the result .
Further, the object is to provide a disk-shaped sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a trace amount of a magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-shaped sample. Rotating means for rotating the holder around its central axis, elevating means for raising and lowering the specimen on the disc-shaped sample holder, cutting means for cutting and removing a part of the specimen, and cutting a part of the specimen An image acquisition means for magnifying the removed portion; a magnetization means for magnetizing the specimen; a magnetic sensor for detecting a magnetic field generated from the magnetized labeled specimen; and a magnetic shield surrounding the magnetic sensor; The plurality of disk-type sample holders are configured to be sequentially inserted from the outside to the inside of the magnetic shield by the rotation of the disk-type sample holder, and the magnetization means is the labeling The body is magnetized outside the magnetic shield, the magnetic sensor detects the magnetic field generated from the magnetized labeled specimen inside the magnetic shield, and the detection of the magnetic field and the magnetization are performed in parallel. The specimen is continuously cut and the magnetic field generated from the labeled specimen magnetized by the magnetic sensor before and after the cutting is measured, and the specimen is measured when the magnetic field is measured before and after the cutting process. After cutting, perform cutting, lower the specimen before measurement, return the specimen to the position before cutting, perform measurement after cutting, then raise the specimen again to the position after cutting, This is achieved by a trace magnetic drug detection device characterized in that the specimen after cutting is magnetized.

  This is achieved by providing the disk-type sample holder with cooling means for freezing and solidifying the specimen of the cell tissue.

  This is achieved by configuring the cooling means with a liquid refrigerant.

  This is achieved by configuring the cooling means with a liquid refrigerant cooled with dry ice.

  This is achieved by configuring the cooling means with a liquid refrigerant cooled by an electronic cooling means.

  This is achieved by configuring the cooling means with a non-magnetic heat storage material cooled by an electronic cooling means.

  This is achieved by configuring the cooling means with a non-magnetic holder cooled by an electronic cooling means.

  This is achieved by providing a cleaning means for removing and collecting a section generated after cutting by the cutting means.

Further, the object is to provide a disk-shaped sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a trace amount of a magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-shaped sample. A rotating step of rotating the holder about its central axis, a lifting step of raising and lowering the specimen on the disk-type sample holder, a cutting step of cutting and removing a part of the specimen, and a part of the specimen being cut An image acquisition process for magnifying the removed portion, a magnetization process for magnetizing the specimen, and a process for detecting a magnetic field generated from the magnetized labeled specimen are performed continuously or programmed. magnetic a magnetic agent detection method, in the step of detecting the magnetic field, to the results measured before and after the cutting process, characterized in that the calculation step of calculating a difference is made to It is achieved by the agent detection methods.

Further, the object is to provide a disk-shaped sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a trace amount of a magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-shaped sample. A rotating step of rotating the holder about its central axis, a lifting step of raising and lowering the specimen on the disk-type sample holder, a cutting step of cutting and removing a part of the specimen, and a part of the specimen being cut An image acquisition process for magnifying the removed portion, a magnetization process for magnetizing the specimen, and a process for detecting a magnetic field generated from the magnetized labeled specimen are performed continuously or programmed. a magnetic agent detection method and, in the process to be measured before and after the cutting step, the cutting step after the step of raising the specimen is performed, to lower the sample before step that is subsequently measured To execute the extent, run the process to be measured after cutting back the specimen position before cutting, is subsequently step of raising the specimen again to the position after cutting performed, to magnetize the sample after the cutting magnetization It is achieved by a magnetic drug detection method characterized by comprising a step of performing a step.

  This is achieved by performing, as necessary, a step of combining a rotating step of rotating the disk-shaped sample holder around its central axis and a radial moving step of transferring in the radial direction of the rotation.

  This is achieved by performing a rotation process for rotating the disk-shaped sample holder about its central axis and a turn-back rotation process in the forward and reverse directions as necessary.

  According to the present invention, a minute amount of magnetic fine particles and a SQUID magnetic sensor can be used to detect a minute amount of magnetic fine particles by a magnetic method with high sensitivity and efficiently detecting a two-dimensional and three-dimensional minute concentration distribution. Can provide measurement technology.

  Hereinafter, an embodiment of the present invention will be described with reference to the accompanying drawings.

  In the trace magnetic drug detection device and the magnetic drug detection method described in the embodiments of the present invention, a trace magnetic drug as shown in FIG. 1 is applied. In addition, in the trace magnetic drug detection device according to the present embodiment, it is desirable that the constituent elements used in the vicinity of performing magnetic field detection be made of a nonmagnetic material from the viewpoint of suppressing magnetic noise from being mixed into the measurement.

FIG. 3 is a cross-sectional view showing the configuration of the trace magnetic drug detection device of the present embodiment, and shows the configuration of the SQUID and the cooling system.
In FIG. 3, for example, a SQUID sensor 61 made of an yttrium-based high-temperature superconducting material reduces the distance between the end surface of the test cell tissue 63, which is a cell tissue sample in the disc-shaped sample holder 62, and SQUID1, and generates a magnetic signal. In order to increase the detection sensitivity and the spatial resolution, the SQUID 61 is arranged close to the disc-shaped sample holder 62. In order to place the SQUID 61 close to the disc-shaped sample holder 62, the SQUID 61 is not cooled directly by the liquid nitrogen 64 but is indirectly cooled via the copper rod 65 and the sapphire rod 66 having high thermal conductivity. The structure to be adopted is adopted. By interposing the sapphire rod 66 between the SQUID 61 and the copper rod 65, there is an effect of reducing the influence of magnetic noise generated from the copper rod 65.

The outer tank 67a and the inner tank 67b of the vacuum heat insulating container 67 are made of a nonmagnetic material such as SUS or FRP. Further, in order to reduce the distance between the minute amount of magnetic fine particles in the disc-shaped sample holder 62 and the SQUID 61, the SQUID 61 and the end surface of the test cell tissue 63 are separated by a thin sapphire window 68 having a thickness of 1 mm or less. . In order to detect magnetic fields with high efficiency,
The distance between the SQUID 61 and the inner bottom surface of the end surface of the test cell tissue 63 is preferably 1.5 mm or less. Liquid nitrogen 64 is replenished from a replenishment pipe 69. A space between the outer tank 67a and the inner tank 67b is an adiabatic vacuum space.

The test cell tissue 63 is inserted so as to be slidable in a cylindrical sapphire support cylinder 70 fixed to a disc-shaped sample holder 62 made of a nonmagnetic material. As a result, the test cell tissue 63 is controlled to move up and down in the cylindrical sapphire support cylinder 70 by the vertical rod 73 whose end is supported by the vertical movement body 72 supported by the vertical movement mechanism 71. Since the disk-shaped sample holder 62 is filled with a refrigerant 74 such as alcohol or antifreeze cooled with dry ice, the cylindrical sapphire support cylinder 70 is cooled to a low temperature of −80 ° C. by the cold heat. The test cell tissue 63 is frozen and solidified.

  The inner surface of the sapphire support cylinder 70 is preferably coated with a material having a low friction coefficient such as a fluororesin in order to reduce the friction coefficient with the test cell tissue 63. The upper and lower rods 73 and the disk-shaped sample holder 62 are airtightly integrated with a bellows 75 so that the refrigerant 74 does not leak. Further, the lower part of the sapphire support cylinder 70 and the disc-shaped sample holder 62 are airtightly integrated with an adhesive. The cylindrical sapphire support cylinder 70 is desirably provided with an introduction port so that the coolant 74 enters the inside thereof to cool the examination cell tissue 63 better.

The disk type sample holder 62 is controlled to rotate by a rotating shaft 76. The disc-shaped sample holder 62 and the rotating shaft 76 are airtightly integrated with an adhesive or the like. The rotation shaft 76 is rotationally controlled by, for example, an ultrasonic motor (not shown) with little magnetic noise in the support moving body 78 that can move on the support base 77. Further, the vertical moving body 72 is moved up and down by the vertical moving mechanism 71 around the disk type sample holder 62 and with the rotary shaft 76 as a core axis. Each control is operated from the electric calculation control device 79 through the signal line 80.

  In order to detect the rotational position of the disk-shaped sample holder 62, a marker (not shown) attached to the periphery of the disk-shaped sample holder 62 is irradiated with laser light from the position detecting device 81, and the reflected light is reflected on the position detecting device. The information is read by the optical sensor, and the information is transmitted to the electric calculation control device 79 through the signal line 82.

  Further, the periphery of the SQUID 61 is surrounded by a double magnetic shield 83 of an outer magnetic shield wall 83a and an inner magnetic shield wall 83b made of permalloy or the like in order to shield magnetic noise in the inspection environment, and part of the disk is a disk. The mold sample holder 62 is opened so that it can enter and exit. A rotary cutter 85, which is a cutting means such as a sharp nonmagnetic cutter blade 84 made of a material such as a rotating ceramic, is provided outside the magnetic shield 83 for the purpose of cutting the end face of the test cell tissue 3. Is provided. The rotary cutter 85 is a shaft 88 supported by a motor 86 and a vertical lift mechanism 87, and its rotational speed and position are operated through a support arm 89 through a signal line 80 from an electric arithmetic control device 79.

  In the end region of the rotary cutter 85, a cleaner 90 that removes and collects a cut piece of the test cell tissue 63 attached to the cutter blade by using, for example, an air blowing suction mechanism or the like, and rotates the support arm 91 to the support arm 89. Is provided. The operation is operated through a signal line 80 from the electric arithmetic control device 79. In addition, the cut pieces of the test cell tissue 63 that have fallen on the upper cover 92 of the disk-type sample holder 62 during the cutting are similarly removed by a cleaner 93 that is removed and collected by, for example, an air blowing suction mechanism. The operation is operated by the electric arithmetic control device 79 via the signal line 94.

  In this way, the test cell tissue 63 solidified on the disc-shaped sample holder 62 is sliced off by the cutter blade 84 to expose a surface parallel to the rotation direction. After the end surface of the test cell tissue 63 is cut to a predetermined thickness, for example, 0.5 mm, the end surface is photographed with an optical microscope 95 at a predetermined rotational position of the disc-shaped sample holder 62, and the image data is, for example, The signal is converted into a signal by the digital camera 96, transmitted to the electric arithmetic control device 79 through the signal line 97, and stored as electronic image data. The end surface portion of the test cell tissue 63 has an appropriate illuminance necessary for photographing with illumination 98 constituted by a light emitting diode or the like, and the control is operated by the electric arithmetic control device 79 via the signal line 99. In this way, the end face is photographed with an optical microscope, and for example, digitized first image data is acquired.

  A permanent magnet for magnetization 100 for magnetizing fine magnetic particles constituting a small amount of magnetic drug contained in the test cell tissue 63 is attached to a shaft 102 supported by an upper limit elevating mechanism 101 and is connected via a signal line 103. The raising / lowering position of the lever is controlled by the electric calculation control device 79. Here, the movement of the trace amount 10 contained in the test cell tissue 63 is moved to the permanent magnet 100 disc sample holder 62 side, and the disc type sample holder rotates so that the cut end surface portion of the test cell tissue 63 is moved to the permanent magnet. Move it directly below. In this way, the fine magnetic particles constituting the minute amount of magnetic drug contained in the test cell tissue 63 are magnetized.

Thereafter, the permanent magnet 100 for excitation is once moved away from the disk-shaped sample holder 62 and the SQUID 61 side to a position where the magnetic noise of the SQUID 61 does not occur. Thereafter, the disc-shaped sample holder 2 is rotated to move the test cell tissue 3 directly below the SQUID 61 surrounded by the magnetic shield 23. Here, the end surface portion of the test cell tissue 63 is rotated and moved directly below the SQUID 61, and the amount of magnetic flux change across the SQUID 61 is precisely measured. At this time, in order to increase the measurement accuracy, the disk-shaped sample holder 62 is rotated directly under the SQUID 61 a plurality of times, for example, 10 times, or reciprocally rotated by forward or reverse operation to obtain a plurality of data, and the data is electrically calculated by the signal line 104. The data is transmitted to the control device 79, the average value is calculated in the electric calculation control device 79, and the data is stored as the first magnetic flux amount a.

  Thereafter, the disc-shaped sample holder 62 is rotated to move the test cell tissue 63 to the outside of the magnetic shield, the vertical moving mechanism 71 is moved up by a predetermined rising amount, for example, 0.5 mm, and the vertical rod 73 is moved to sapphire. The test cell tissue 63 in the support tube 70 is raised by 0.5 mm. The ascending portion is cut by the cutter blade 84, and the vertical moving mechanism 71 lowers the vertical moving body 72 by 0.5 mm before the cutting, and the vertical rod 73 lowers the inspection cell tissue 63 in the sapphire support cylinder 70 to 0. Lower 5mm.

  Next, the disc-shaped sample holder 62 is rotated as it is without being magnetized, and the test cell tissue 63 is moved directly below the SQUID 61 surrounded by the magnetic shield, and the same operation as that for obtaining the first magnetic flux amount a is performed. Thus, data of the first magnetic flux amount b of the end surface portion of the inspection cell tissue 63 is acquired.

  Next, a first magnetic flux difference amount obtained by subtracting the first magnetic flux amount b from the first magnetic flux amount a is obtained by the electric calculation control device 79. By this operation, the first image data deleted during the measurement of the first magnetic flux amount “a” to the first magnetic flux amount “b” is calculated as the magnetic flux amount of the test cell tissue end face section deleted.

In order to measure the second magnetic flux amount, the disk-shaped sample holder 62 is rotated to move the test cell tissue 63 out of the magnetic shield, and the vertical moving mechanism 71 moves the vertical moving body 72 by a predetermined amount of 0.5 mm. Raise.

  Next, the cutting operation by the cutter blade 84 is not performed, and the end face is photographed with the optical microscope 95 at the rotation position of the predetermined disc-shaped sample holder 62. The photographed image data is converted into a signal by, for example, a digital camera 96, transmitted to the electric arithmetic control device 79 through the signal line 97, and stored as second image data.

  Next, the upper limit elevating mechanism 101 moves to the permanent magnet 100 disc type sample holder 62 side to rotate the disc type sample holder, and moves the cut end surface portion of the test cell tissue 63 directly below the permanent magnet to check the test cell tissue 63. The fine magnetic particles inside are magnetized.

Thereafter, the permanent magnet 100 for excitation is moved away from the disk-shaped sample holder 62 and the SQUID 61 side to a position where the magnetic noise of the SQUID 1 is not generated. After the movement, the disc-shaped sample holder 62 is rotated so that the test cell tissue 63 is surrounded by the magnetic shield 83.
Move to just below 61. Thereafter, the end surface portion of the test cell tissue 63 is rotated and moved directly below the SQUID 61, and the disk-shaped sample holder 62 is rotated 10 times immediately below the SQUID 61, and reciprocally rotated by forward or reverse operation to obtain a plurality of data. Is transmitted to the electric calculation control device 79, the average value is calculated in the electric calculation control device 79, and the data is stored as the second magnetic flux amount a.

Thereafter, the disc-shaped sample holder 62 is rotated to move the test cell tissue 63 out of the magnetic shield, the vertical moving mechanism 71 is moved up by a predetermined rising amount, for example, 0.5 mm, and the vertical rod 73 is moved up and down. The test cell tissue 63 in the sapphire support cylinder 70 is raised by 0.5 mm, the rise is cut by the cutter blade 84, and the vertical movement mechanism 71 is again raised by the vertical movement mechanism 71 by 0.5 mm. The test cell tissue 63 in the sapphire support cylinder 70 is lowered by 0.5 mm by the upper and lower rods 73.

  Next, the same operation as when the second magnetic flux amount a is acquired by rotating the disc-shaped sample holder 62 as it is without being magnetized and moving the test cell tissue 3 directly below the SQUID 61 surrounded by the magnetic shield. Thus, data of the second magnetic flux amount b of the end surface portion of the test cell tissue 63 is acquired.

  Next, a second magnetic flux difference amount obtained by subtracting the second magnetic flux amount b from the second magnetic flux amount a is obtained by the electric calculation control device 79. By this operation, it is calculated as the magnetic flux amount corresponding to the test cell tissue end face section of the second image data deleted during the measurement of the second magnetic flux amount a to the second magnetic flux amount b. Next, in order to measure the third magnetic flux amount, the disk-shaped sample holder 62 is rotated to move the test cell tissue 63 out of the magnetic shield, and the vertical moving mechanism 71 moves the vertical moving body 72 to a predetermined ascending amount. Raise 0.5mm.

  By repeating this operation, the data of each layer is obtained, the data for each layer stored in the electric calculation control device 79 is digitally processed, and a three-dimensional image of the entire examination cell tissue is formed from the image data. Furthermore, the amount of the magnetic drug alone obtained experimentally separately from the magnetic flux amount data of each layer and the amount of the magnetic magnetic agent calculated from the magnetic flux amount of each cutting layer from the calibration data of the measured magnetic flux amount are obtained by calculation, The concentration distribution can be imaged from the distribution of the trace magnetic drug amount in the examination cell tissue 63, and the concentration distribution of the trace magnetic drug amount can be imaged so as to overlap the optical 3D image. The imaged image is displayed on the monitor 105.

  Further, since the inspection apparatus is operated at a low temperature, moisture in the atmosphere is condensed. Therefore, in order to prevent this, the device is covered with a cover, and air whose humidity has been reduced by the dehumidifying device 106 is blown into the cover 105a through the duct 107, and the inside of the cover 105a is kept slightly higher than the atmospheric pressure outside the cover. In order to prevent condensation.

  As described above, in this embodiment, the position control and continuous measurement of the cell tissue sample can be easily operated, the cell tissue sample is cooled and solidified, the cell tissue sample is sequentially sliced, and images of each section are taken, and the amount of magnetic flux is cut. Since the magnetic mass can be accurately and accurately measured based on the difference between the amount of magnetic flux before and after, the concentration distribution in the thickness direction of the tissue sample can be accurately and efficiently measured. The total amount and concentration distribution of the magnetic drug to be measured are three-dimensionally measured together with the image data, and the concentration distribution can be displayed on the monitor 105 together with the three-dimensional image of the cell tissue sample and visualized.

  In the present embodiment, a refrigerant cooled with dry ice is used to solidify the cell tissue sample, but the same effect can be obtained by using liquid nitrogen or liquid air instead of dry ice.

FIG. 4 is a schematic configuration diagram of an apparatus showing another embodiment.
In the embodiment of FIG. 3, a refrigerant cooled with dry ice is used to freeze and solidify the test cell tissue 63, but in the embodiment of FIG. 4, a Peltier element or the like is used instead of dry ice, and electromagnetic noise is generated. In this configuration, cooling is performed with a small number of electronic refrigeration apparatuses 108. As a result, power is supplied to the electronic refrigeration apparatus 108 through the wiring 109 connected to the electric arithmetic control apparatus 79.

  According to the present embodiment, since the refrigerant 74 can be cooled without supplying dry ice, there is no need to purchase and store a cooling source such as dry ice, and a new effect that makes the operation easier becomes possible. .

FIG. 5 is a schematic configuration diagram of an apparatus showing another embodiment.
In the embodiment of FIG. 4, a liquid refrigerant cooled with dry ice is used to freeze and solidify the test cell tissue 63. However, in the embodiment of FIG. In addition, a non-magnetic heat storage body 111 having a heat storage effect, such as aluminum or copper that can be cooled over time, is arranged. At this time, the lid 110 covers the entire container in a heat-insulating manner, and the disc-shaped sample holder 62 is made of a material having excellent heat insulating properties. The sapphire support cylinder 70 and the heat storage body 111 are thermally integrated with an adhesive or the like.

  According to the present embodiment, since the sapphire support cylinder 70 can be cooled without supplying and replenishing the liquid refrigerant, there is an effect that the operation operation becomes easier.

FIG. 6 is a schematic configuration diagram of an apparatus showing another embodiment.
This embodiment is different from the embodiment of FIG. 5 in that the sapphire support cylinder 70 is directly cooled by the electronic refrigeration apparatus 108 in order to freeze and solidify the test cell tissue 63. The sapphire support cylinder 70 and the electronic refrigeration apparatus 108 are thermally integrated with an adhesive or the like.

  According to the present embodiment, since a liquid refrigerant or a heat storage body is not required, the apparatus configuration is simplified, and a new effect that the apparatus can be further reduced in weight is produced.

  In the above embodiment, the range measured by the SQUID 61 is the same as that when the test cell tissue 63 crosses directly under the SQUID 61 by rotating the disc type sample holder 62 with the disc type sample holder 62 at one point of the support base 77. The amount of magnetic flux was measured by the amount of change in the magnetic field, but the change in magnetic flux between the small regions in the moving circumferential plane of the end surface of the test cell tissue 63 is measured by moving the disc-shaped sample holder 62 in small increments immediately below the SQUID 61. Thus, the circumferential magnetic flux distribution in the rotation direction within the end surface of the test cell tissue 63 can also be measured. In this case, it is possible to measure the radial magnetic flux distribution in the rotational direction in the end surface of the test cell tissue 63 by controlling the movement of the support moving table in the rotational radial direction of the disk-shaped sample holder 62 and performing the same measurement. An effect is produced.

  In the present invention, if the test cell tissue 63 is solidified with a normal temperature solid agent such as agar, a cooling source and a dew condensation prevention structure are not required, and the total amount and concentration distribution of the magnetic drug contained in the cell tissue sample can be obtained more efficiently. It is possible to measure three-dimensionally together with image data and create a density distribution.

  As described above, according to the present invention, the position control and continuous measurement of the cell tissue sample can be easily performed, the cell tissue sample is cooled and solidified, the cell tissue sample is sequentially sliced, and the image of each section is taken, and the amount of magnetic flux is reduced. Since the magnetic mass can be accurately and accurately measured based on the difference value of the magnetic flux before and after cutting, the concentration distribution in the thickness direction and plane direction of the cell tissue sample can be accurately and efficiently measured. The total amount and concentration distribution of the magnetic drug contained in the tissue sample can be measured three-dimensionally together with the image data, and the concentration distribution can be displayed on the monitor 105 together with the three-dimensional image of the cell tissue sample for visualization.

  As described above, in the present invention, a magnet or a magnetic field generating coil is disposed outside a magnetic shield as a means for applying an external magnetic field to a minute amount of magnetic fine particles distributed three-dimensionally in a test cell tissue, and is generated from the minute amount of magnetic fine particles. A magnetic sensor for detecting a magnetic field is disposed inside the magnetic shield. A disk-type sample holder for storing a test cell tissue containing a minute amount of magnetic fine particles is held on the circumference of a disk-type sample holder made of a non-magnetic material.

  The non-magnetic disc type sample holder includes a mechanism for fixing a plurality of disc type sample holders on the circumference. The disc-shaped sample holder can be rotated by a rotation mechanism composed of an ultrasonic motor. The SQUID magnetic sensor is disposed inside the magnetic shield device. A moving mechanism and a position adjusting mechanism for inserting the disk-shaped sample holder into the magnetic shield device are provided.

  In order to detect the rotational position of the disk-shaped sample holder, a mechanism is provided that detects the position by irradiating a marker attached to the periphery of the disk-shaped sample holder with laser light and reading the reflected light with an optical sensor. A part of the magnetic shield has a hole for passing the disk type holder and the above rotating mechanism so that a part of the disk type holder is exposed to the outside of the magnetic shield. During the measurement of the magnetic field generated from a minute amount of magnetic fine particles in a sample placed on the magnet, the minute amount of magnetic fine particles in the sample of the disk-shaped sample holder at a position different from the above position is magnetized by the magnet or the magnetic field generating coil. It is configured to be able to be made.

  On the other hand, the test cell tissue fixed to the data container is formed with a predetermined non-cutting thickness of 1 μm or more by a cutting means such as a sharp non-magnetic cutter blade made of a material such as ceramic rotating outside the magnetic shield. The thickness is cut off, and the end face is photographed with an optical microscope at the rotation position of a predetermined disc-shaped sample holder. The image data is stored in an electronic image such as a personal computer, and the processing device, cutting, and rotation drive control are performed. Is configured to be controlled by an electrical control device. Here, it is configured such that the test cell tissue can be frozen, for example, by solidification means such as a refrigeration apparatus and the solid phase state can be maintained.

  In this way, the test cell tissue solidified on the disk type sample holder is sliced by the cutting means to expose a surface parallel to the rotation direction, and the test sample tissue is rotated by rotating the disk type sample holder. Is moved directly under the optical microscope, and first, the end face is photographed with the optical microscope to obtain, for example, digitized first image data. In addition, the carter part of the cutting means is moved into the wiping container after the cutting operation, and the cutting pieces on the blade edge part are sufficiently removed. After that, the permanent magnet for excitation is moved to the disk-type sample holder side, and the disk-type sample holder is rotated to move the test cell tissue directly under the permanent magnet to be magnetized. Move away from the disk-shaped sample holder. Thereafter, the disc-shaped sample holder is rotated to move the test cell tissue directly below the SQUID element surrounded by the magnetic shield, and data on the first magnetic flux amount a at the end surface portion of the test cell tissue is acquired.

  Thereafter, the disk-shaped sample holder is rotated to move the test cell tissue out of the magnetic shield, raised by a predetermined distance by a minute lifting / lowering means, and sliced and removed by the cutting means. Next, this time, the disk-shaped sample holder is rotated as it is to move the examination cell tissue directly below the SQUID element surrounded by the magnetic shield, and data on the first magnetic flux amount b at the end portion of the examination cell tissue is acquired. The first image obtained by subtracting the first magnetic flux amount b from the first magnetic flux amount a by the electronic computing means is deleted during the measurement of the first magnetic flux amount b from the first magnetic flux amount a. Examination of data Calculated as the amount of magnetic flux at the end surface portion of the tissue. By repeating this operation, the data of each layer is obtained, and the data for each layer is imaged, whereby the concentration distribution of the magnetic flux amount can be imaged so as to overlap the optical 3D image.

The trace magnetic drug detection device of the present invention has a SQUID that detects a magnetic field generated from a minute amount of magnetic fine particles with high sensitivity, a cryostat that cools the SQUID, and a drive circuit that drives the SQUID, and collects and displays the output of the SQUID. And a device for performing addition processing. Also,
A SQUID temperature increasing light source is provided for heating the SQUID to a superconducting transition temperature or higher by a laser or halogen lamp light source to remove magnetic flux traps that cause magnetic noise. The removal of the magnetic flux trap is executed prior to the measurement of the magnetic field.

  In the trace magnetic drug detection device of the present invention, when the distance between the detection coil surface of the magnetic sensor and the end surface of the test cell tissue is d, an adjacent sample or a magnetic part is placed at an interval of d√2 or more. In other words, interference between magnetic signals emitted from adjacent members is suppressed by setting the gap distance between adjacent samples or magnetized parts to be d√2 or more. In addition, a SQUID temperature raising light source is provided in order to quickly remove magnetic flux traps that cause SQUID magnetic noise. An ultrasonic motor with low magnetic noise was used as the rotating mechanism for the disk-shaped sample holder. To more accurately measure the magnetic signal of a minute amount of magnetic particles, rotate the disk-shaped sample holder multiple times, and measure the magnetic signal of a minute amount of magnetic particles every rotation while the sample is rotating. , So that the averaging can be performed.

  The trace magnetic drug detection device of the present invention is summarized as follows. In order to facilitate the position control and continuous measurement of the cell tissue sample, a disk-type sample holder having a solidification means that can be rotationally controlled by a rotating mechanism was used. A detection method for measurement was used in which a cell tissue sample was sequentially sliced, and the amount of magnetic flux for each slice was determined from the difference value of the amount of magnetic flux before and after cutting.

  As described above, the concentration distribution in the thickness direction of the cell tissue sample can be efficiently measured in the measurement of a minute amount of magnetic fine particles.

It is a figure explaining the structure of the magnetic chemical | medical agent using a trace amount magnetic particle. It is a figure explaining the structural example of the apparatus which measures the magnetic flux amount of the conventional magnetic fine particle using SQUID. It is a figure explaining the structure of the trace magnetic chemical | medical agent detection apparatus which becomes one Example of this invention. It is a figure explaining the structure of the trace magnetic chemical | medical agent detection apparatus which becomes another Example of this invention. It is a figure explaining the structure of the trace magnetic chemical | medical agent detection apparatus which becomes another Example of this invention. It is a figure explaining the structure of the trace magnetic chemical | medical agent detection apparatus which becomes another Example of this invention.

Explanation of symbols

4 Rotating mechanism 61 High-temperature superconducting SQUID
62 Disk type sample holder 63 Test cell tissue 64 Liquid nitrogen 65 Copper rod 66 Sapphire rod 67 Vacuum heat insulating container 68 Sapphire window 70 Sapphire support cylinder 71 Vertical moving mechanism 72 Vertical moving body 73 Vertical rod 74 Refrigerant 76 Rotating shaft 79 Electric calculation control device 83, 83a, 83b Magnetic shield 84 Cutter blade 87 Vertical elevating mechanism 90 Cleaner 95 Microscope 96 Digital camera 100 Permanent magnet 101 Upper limit elevating mechanism 105 Monitor 105a Cover 106 Dehumidifying device 108 Electronic refrigerating device 109 Wiring 111 Heat storage body

Claims (13)

A sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a minute amount of magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-type sample holder on the center axis Rotating means for rotating around, lifting / lowering means for moving the specimen up and down on the disc-shaped sample holder, cutting means for cutting and removing a part of the specimen, and enlarged photographing of a part where the specimen is cut and removed Image acquisition means, magnetizing means for magnetizing the specimen, a magnetic sensor for detecting a magnetic field generated from the magnetized labeled specimen , and a magnetic shield surrounding the magnetic sensor,
The plurality of disk-type sample holders are configured to be sequentially inserted from the outside of the magnetic shield by the rotation of the disk-type sample holder, and the magnetizing means magnetizes the labeled specimen outside the magnetic shield, The magnetic sensor detects a magnetic field generated from the magnetized labeled specimen inside the magnetic shield, the detection of the magnetic field and the magnetization are performed in parallel, and the specimen is continuously cut and cut. While configured to measure the magnetic field generated from the labeled specimen magnetized by the magnetic sensor before and after ,
When detecting the said magnetic field, it comprised so that the difference might be calculated from the result measured before and after the said cutting process, The trace magnetic chemical | medical agent detection apparatus characterized by the above-mentioned. A sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a minute amount of magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-type sample holder on the center axis Rotating means for rotating around, lifting / lowering means for moving the specimen up and down on the disc-shaped sample holder, cutting means for cutting and removing a part of the specimen, and enlarged photographing of a part where the specimen is cut and removed Image acquisition means, magnetizing means for magnetizing the specimen, a magnetic sensor for detecting a magnetic field generated from the magnetized labeled specimen, and a magnetic shield surrounding the magnetic sensor,
  The plurality of disk-type sample holders are configured to be sequentially inserted from the outside of the magnetic shield by the rotation of the disk-type sample holder, and the magnetizing means magnetizes the labeled specimen outside the magnetic shield, The magnetic sensor detects a magnetic field generated from the magnetized labeled specimen inside the magnetic shield, the detection of the magnetic field and the magnetization are performed in parallel, and the specimen is continuously cut and cut. While configured to measure the magnetic field generated from the labeled specimen magnetized by the magnetic sensor before and after,
  When measuring the magnetic field before and after the cutting process, perform cutting after raising the specimen, then lower the specimen before measuring, and return the specimen to the position before cutting to perform measurement after cutting The specimen is then raised again to the position after cutting, and the specimen after cutting is magnetized. In the trace magnetic chemical | medical agent detection apparatus of Claim 1 or 2 ,
An apparatus for detecting a minute amount of magnetic drug, comprising a cooling means for freezing and solidifying the specimen of the cell tissue in the disk-type sample holder. In the trace magnetic chemical | medical agent detection apparatus of Claim 3 ,
A trace magnetic drug detection device, wherein the cooling means is composed of a liquid refrigerant. In the trace magnetic chemical | medical agent detection apparatus of Claim 3 ,
An apparatus for detecting a minute amount of magnetic drug, wherein the cooling means is composed of a liquid refrigerant cooled by dry ice. In the trace magnetic chemical | medical agent detection apparatus of Claim 3 ,
An apparatus for detecting a minute amount of magnetic drug, wherein the cooling means is composed of a liquid refrigerant cooled by an electronic cooling means. In the trace magnetic chemical | medical agent detection apparatus of Claim 3 ,
An apparatus for detecting a minute amount of magnetic drug, wherein the cooling means is composed of a non-magnetic heat storage material cooled by an electronic cooling means. In the trace magnetic chemical | medical agent detection apparatus of Claim 3 ,
An apparatus for detecting a minute amount of magnetic drug, wherein the cooling means is composed of a non-magnetic holder cooled by an electronic cooling means. In the trace magnetic chemical | medical agent detection apparatus of Claim 1 or 2 ,
A trace magnetic drug detection device comprising a cleaning means for removing and collecting a section generated after cutting by the cutting means. A sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a minute amount of magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-type sample holder on the center axis A rotating step for rotating the sample around, a lifting step for moving the sample up and down on the disc-shaped sample holder, a cutting step for cutting and removing a part of the sample, and a magnified image of a portion obtained by cutting and removing a part of the sample A magnetic drug detection method comprising: performing an image acquisition step, a magnetization step of magnetizing the specimen, and a step of detecting a magnetic field generated from the magnetized labeled specimen, continuously or programmed. Because
A magnetic drug detection method comprising a step of calculating a difference from results measured before and after the cutting step in the step of detecting the magnetic field. A sample holder for holding a sample holder for storing a specimen of a cellular tissue containing a minute amount of magnetic drug containing minute magnetic fine particles combined with a non-magnetic substance on the circumference, and the disk-type sample holder on the center axis A rotating step for rotating the sample around, a lifting step for moving the sample up and down on the disc-shaped sample holder, a cutting step for cutting and removing a part of the sample, and a magnified image of a portion obtained by cutting and removing a part of the sample A magnetic drug detection method comprising: performing an image acquisition step, a magnetization step of magnetizing the specimen, and a step of detecting a magnetic field generated from the magnetized labeled specimen, continuously or programmed. Because
In the process that is measured before and after the cutting process, the cutting process is executed after the process of raising the specimen, and the process of lowering the specimen is executed before the process to be measured thereafter, and the specimen is returned to the position before cutting and after the cutting. A method of detecting a magnetic drug, comprising performing a step of measuring, then performing a step of raising the specimen again to a position after cutting, and performing a magnetization step of magnetizing the specimen after cutting. The magnetic drug detection method according to claim 10 or 11 ,
A magnetic drug detection method comprising a step of combining a rotating step of rotating the disc-shaped sample holder about its central axis and a radial moving step of transferring the disc-shaped sample holder in the radial direction of the rotation as necessary. The magnetic drug detection method according to claim 10 or 11 ,
A magnetic drug detection method comprising: a rotating step of rotating the disk-shaped sample holder about its central axis and a turning-back rotating step in the forward and reverse directions as necessary.

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