US20210208088A1 - Magnetic holder for immunoelectron microscopy grids - Google Patents
Magnetic holder for immunoelectron microscopy grids Download PDFInfo
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- US20210208088A1 US20210208088A1 US17/140,043 US202117140043A US2021208088A1 US 20210208088 A1 US20210208088 A1 US 20210208088A1 US 202117140043 A US202117140043 A US 202117140043A US 2021208088 A1 US2021208088 A1 US 2021208088A1
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- nickel
- holder
- frame
- arm
- magnet
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2202—Preparing specimens therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2204—Specimen supports therefor; Sample conveying means therefore
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/30—Accessories, mechanical or electrical features
- G01N2223/309—Accessories, mechanical or electrical features support of sample holder
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/60—Specific applications or type of materials
- G01N2223/612—Specific applications or type of materials biological material
Definitions
- the present invention relates to a holder for carrying nickel grids with biological ultrasections used in immunoelectron microscope sample preparation technology, in particular to a holder for carrying the nickel grids with a high throughput during rinsing, antigen-antibody labeling and staining.
- a nickel grid is a necessary ultrasections carrier applied to biological sample preparation technology of immunoelectron microscope.
- the nickel grid comprises two parts: one is a nickel ring with a rim width of about 0.2 mm, and another is hollow grids of different sizes ranging from 50-200 meshes inside the nickel ring.
- the thickness of the whole nickel grid is about 18 micrometers.
- the entire operation comprises many steps such as rinsing (multiple times), blocking liquid incubation, primary antibody incubation, rinsing (multiple times), secondary antibody incubation, rinsing (multiple times), fixative, rinsing (multiple times), and staining.
- steps such as rinsing (multiple times), blocking liquid incubation, primary antibody incubation, rinsing (multiple times), secondary antibody incubation, rinsing (multiple times), fixative, rinsing (multiple times), and staining.
- steps such as rinsing (multiple times), blocking liquid incubation, primary antibody incubation, rinsing (multiple times), secondary antibody incubation, rinsing (multiple times), fixative, rinsing (multiple times), and staining.
- the transfer of the nickel grid in each step of the operation is realized by using a pair of tweezers to grip the nickel ring with a rim
- the tweezers touch the grid inner area of the nickel grid in this process, the tweezers can easily touch sections on the grid or the Formvar membrane or carbon membrane adheres to the surfaces of the grids, thereby directly or indirectly damaging the sections loading on the nickel grid, thus affecting following observation .
- these unexpected background are irregularly mixed with the secondary antibody colloidal gold particles of only 6-15 nm in the immunolabeling, which seriously affects the imaging effect.
- usually one immunolabeling experiment needs to simultaneously process multiple sets of sections for labeling under different test conditions; and the number of sections processed in batches is large. This multiplies the steps of transferring sections, the error rate, and the operation time.
- CN105910875A discloses a device which can perform batch staining on the copper grids.
- the device comprises a main body for fixing the copper grids with sample sections and an auxiliary body for fixing and supporting the main body.
- the main body comprises soft layers and a hard layer covered by the soft layers, wherein the cross section of the hard layer is arc-shaped or angle-shaped.
- the copper grid is fixed in the device by the soft layers at the two sides of an arc-shaped opening or an angle-shaped opening.
- the above device is mainly used for the copper grid.
- the copper grid has a high hardness and can be easily inserted into a wax layer; but a nickel grid has a low hardness, and is difficult to insert it into a wax layer.
- the device is only suitable for a single staining operation. If the device is used to load a batch of copper grids and continuously transfer the grids in liquid of multiple steps, it will carry too much residual liquid from the previous steps.
- a main technical problem solved by the present invention is to provide a device that can be used for batch operation (such as rinsing, immunolabeling, dyeing, etc.) on nickel grids and can reduce the amount of residual liquid carried by the nickel grids in the process of continuously transferring the nickel grids in liquid of multiple steps.
- the present invention provides a magnetic holder for immunoelectron microscopy grids, comprising a frame, magnets and a hydrophobic layer
- the frame comprises at least one arm made of a solid material, the arm has a hollow interior and axisymmetric arc-shaped grooves located at an upper end and a lower end of an outer surface of the arm and close to the hollow interior, the magnets are detachably arranged below the frame, and the hydrophobic layer adheres to the outer surface of the magnetic holder.
- the frame comprises at least two arms made of a solid material, the frame further comprises a connecting part, each of the arms has a hollow interior, the magnets are disposed in the hollow interiors of the arms, the connecting part is used for connecting the arms, and the hydrophobic layer adheres to an outer surface of the magnetic holder.
- the beneficial technical effects of the present invention the uniform transfer of batches of sample-loaded nickel grids in different reaction solutions is realized.
- the present invention not only can the reaction time of the section in each of the sample-loaded nickel grids be unified, but also greatly reduce the mechanical damage to the grids, and it can also reduce the amount of liquid carried out by nickel grids during the continuous replacement of the nickel net between different liquids to almost zero, which greatly improves the success rate of immunolabeling at the sample preparation level.
- FIG. 1 is a schematic diagram of a magnetic holder with a frame located at one side of a magnet.
- FIG. 2 is a schematic diagram of a magnetic holder with a magnet located between two adjacent arms.
- FIG. 3 is a schematic diagram of a magnetic holder with magnets located inside arms, showing a A-A′ cross section.
- a magnetic holder for nickel grids with a frame located entirely at one side of a magnet comprises the frame 1 , the magnet 2 and a hydrophobic layer 3 .
- the frame 1 comprises at least one arm 101 with a hollow interior.
- the arms 101 are made of a solid material. The distance between these hollow interiors of the arms is 2.6 mm.
- a plurality of axisymmetric arc-shaped grooves are provided in positions located at an upper end and a lower end of an outer surface of the arms 101 and close to the hollow interiors.
- the openings of the arc-shaped grooves all face the opposite sides, and the two symmetrical arc-shaped grooves can just accommodate the nickel ring on the outside of one nickel grid.
- the magnet 2 is a magnetic iron detachably arranged below the frame 1 .
- the hydrophobic layer 3 adheres to the outer surfaces of the frame 1 and the magnet 2 .
- a magnetic holder for immunoelectron microscopy grids with a magnet located between two adjacent arms comprises a frame 1 , the magnet 2 and a hydrophobic layer 3 .
- the frame 1 comprises at least two axisymmetric arms 101 .
- the distance between the two adjacent arms 101 is 2.6 mm to 3 mm.
- the thickness of one arm 101 is not less than 0.1 mm.
- Each of the arms 101 is made of a solid material such as iron, cobalt or nickel, which are easily magnetizable.
- the hydrophobic layer 3 adheres to the outer surface of each of the arms 101 .
- the magnet 2 is a magnetic iron with a length and a width being both smaller than that of the arms 101 .
- the magnet 2 is located between the two adjacent arms 101 , and connects and magnetizes the arms 101 .
- the hydrophobic layer 3 adheres to the outer surface of the magnet 2 .
- a magnetic holder for immunoelectron microscopy grids with magnets located inside arms comprises a frame 1 , the magnets 2 and a hydrophobic layer 3 .
- the frame 1 comprises at least two axisymmetric arms 101 and a connecting part 102 .
- the overall thickness of one arm 101 is not less than 0.1 mm.
- the interiors of the arms are hollow, and the exteriors of the arms are made of a solid material.
- the arms 101 are connected by the connecting part 102 .
- the distance between the two adjacent arms 101 is 2.6 to 3.3 mm.
- the magnets 2 are magnet iron located in the hollow interiors of the arms 101 .
- the hydrophobic layer 3 adheres to the outer surface of the frame 1 .
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
Description
- This application claims the priority of Chinese Patent Application No. 202010015283.2, filed on Jan. 7, 2020, the entire disclosure of which is incorporated herein by reference.
- The present invention relates to a holder for carrying nickel grids with biological ultrasections used in immunoelectron microscope sample preparation technology, in particular to a holder for carrying the nickel grids with a high throughput during rinsing, antigen-antibody labeling and staining.
- A nickel grid is a necessary ultrasections carrier applied to biological sample preparation technology of immunoelectron microscope. The nickel grid comprises two parts: one is a nickel ring with a rim width of about 0.2 mm, and another is hollow grids of different sizes ranging from 50-200 meshes inside the nickel ring. The thickness of the whole nickel grid is about 18 micrometers. There is an organic Formvar layer on the front of the nickel grid. Some laboratories spray a carbon membrane of few nanometers thickness on the outer surface of the Formvar membrane which is used to reduce the non-specific adsorption capacity of immune colloidal gold and to provide stronger support for sections. The immunolabeling operation of biological samples is performed on the nickel grid. The entire operation comprises many steps such as rinsing (multiple times), blocking liquid incubation, primary antibody incubation, rinsing (multiple times), secondary antibody incubation, rinsing (multiple times), fixative, rinsing (multiple times), and staining. Usually the transfer of the nickel grid in each step of the operation is realized by using a pair of tweezers to grip the nickel ring with a rim width of about 0.2 mm around the nickel grid. If the tweezers touch the grid inner area of the nickel grid in this process, the tweezers can easily touch sections on the grid or the Formvar membrane or carbon membrane adheres to the surfaces of the grids, thereby directly or indirectly damaging the sections loading on the nickel grid, thus affecting following observation . In addition, in the process of transferring the nickel grid, there is often residual liquid that was not cleaned in the previous step in the gap of the tweezers and carried into the next reaction solution, which can easily lead to section contamination (i.e. unexpected background) Under high-resolution observation of an electron microscope, these unexpected background are irregularly mixed with the secondary antibody colloidal gold particles of only 6-15 nm in the immunolabeling, which seriously affects the imaging effect. Furthermore, usually one immunolabeling experiment needs to simultaneously process multiple sets of sections for labeling under different test conditions; and the number of sections processed in batches is large. This multiplies the steps of transferring sections, the error rate, and the operation time.
- In order to achieve the unification of the stain time of each section, there is a device for copper grids batch staining in the prior art. CN105910875A discloses a device which can perform batch staining on the copper grids. The device comprises a main body for fixing the copper grids with sample sections and an auxiliary body for fixing and supporting the main body. The main body comprises soft layers and a hard layer covered by the soft layers, wherein the cross section of the hard layer is arc-shaped or angle-shaped. The copper grid is fixed in the device by the soft layers at the two sides of an arc-shaped opening or an angle-shaped opening.
- The above device is mainly used for the copper grid. The copper grid has a high hardness and can be easily inserted into a wax layer; but a nickel grid has a low hardness, and is difficult to insert it into a wax layer. In addition, the device is only suitable for a single staining operation. If the device is used to load a batch of copper grids and continuously transfer the grids in liquid of multiple steps, it will carry too much residual liquid from the previous steps.
- A main technical problem solved by the present invention is to provide a device that can be used for batch operation (such as rinsing, immunolabeling, dyeing, etc.) on nickel grids and can reduce the amount of residual liquid carried by the nickel grids in the process of continuously transferring the nickel grids in liquid of multiple steps.
- In order to solve the above technical problem, the present invention provides a magnetic holder for immunoelectron microscopy grids, comprising a frame, magnets and a hydrophobic layer
- As a preferred structure of the present invention, the frame comprises at least one arm made of a solid material, the arm has a hollow interior and axisymmetric arc-shaped grooves located at an upper end and a lower end of an outer surface of the arm and close to the hollow interior, the magnets are detachably arranged below the frame, and the hydrophobic layer adheres to the outer surface of the magnetic holder.
- As a preferred structure of the present invention, the frame comprises at least two arms made of a solid material, the frame further comprises a connecting part, each of the arms has a hollow interior, the magnets are disposed in the hollow interiors of the arms, the connecting part is used for connecting the arms, and the hydrophobic layer adheres to an outer surface of the magnetic holder.
- The beneficial technical effects of the present invention: the uniform transfer of batches of sample-loaded nickel grids in different reaction solutions is realized. Through the present invention, not only can the reaction time of the section in each of the sample-loaded nickel grids be unified, but also greatly reduce the mechanical damage to the grids, and it can also reduce the amount of liquid carried out by nickel grids during the continuous replacement of the nickel net between different liquids to almost zero, which greatly improves the success rate of immunolabeling at the sample preparation level.
- The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. The drawings are used to explain the present invention together with the following exemplar examples, but do not constitute a limitation to the present application. In the drawings:
-
FIG. 1 is a schematic diagram of a magnetic holder with a frame located at one side of a magnet. -
FIG. 2 is a schematic diagram of a magnetic holder with a magnet located between two adjacent arms. -
FIG. 3 is a schematic diagram of a magnetic holder with magnets located inside arms, showing a A-A′ cross section. - The specific embodiments of the present invention will be described in further detail below in conjunction with the drawings and embodiments. The following examples are used to illustrate the present invention, but not to limit the scope of the present invention.
- As shown in
FIG. 1 , a magnetic holder for nickel grids with a frame located entirely at one side of a magnet comprises theframe 1, themagnet 2 and ahydrophobic layer 3. Theframe 1 comprises at least onearm 101 with a hollow interior. Thearms 101 are made of a solid material. The distance between these hollow interiors of the arms is 2.6 mm. A plurality of axisymmetric arc-shaped grooves are provided in positions located at an upper end and a lower end of an outer surface of thearms 101 and close to the hollow interiors. The openings of the arc-shaped grooves all face the opposite sides, and the two symmetrical arc-shaped grooves can just accommodate the nickel ring on the outside of one nickel grid. Themagnet 2 is a magnetic iron detachably arranged below theframe 1. Thehydrophobic layer 3 adheres to the outer surfaces of theframe 1 and themagnet 2. - Operation: separating the
frame 1 and themagnet 2, placing the outer rings of the upper sides and lower sides of the plurality ofnickel grids 4 respectively in the symmetrical arc-shaped grooves of thearms 101, recombining theframe 1 loaded thenickel grids 4 and themagnet 2, and carrying out the immunolabeling operation. After the immunolabeling is completed, separating theframe 1 and themagnet 2 again. Finally taking out thenickel grids 4. Thenickel grids 4 are fixed by the magnetic force of themagnet 2 under theframe 1 to achieve the batch staining for thenickel grids 4. Thenickel grids 4 can be prevented from carrying too much residual liquid in the process of transferring between different reaction liquids by the hydrophobic effect of the hydrophobic layer. By separating theframe 1 from themagnet 2, it is possible to avoid the interference of the magnetic force on the nickel grids when the nickel grids are put into the frame or taken out of the frame. - As shown in
FIG. 2 , a magnetic holder for immunoelectron microscopy grids with a magnet located between two adjacent arms comprises aframe 1, themagnet 2 and ahydrophobic layer 3. Theframe 1 comprises at least twoaxisymmetric arms 101. The distance between the twoadjacent arms 101 is 2.6 mm to 3 mm. The thickness of onearm 101 is not less than 0.1 mm. Each of thearms 101 is made of a solid material such as iron, cobalt or nickel, which are easily magnetizable. Thehydrophobic layer 3 adheres to the outer surface of each of thearms 101. Themagnet 2 is a magnetic iron with a length and a width being both smaller than that of thearms 101. Themagnet 2 is located between the twoadjacent arms 101, and connects and magnetizes thearms 101. Thehydrophobic layer 3 adheres to the outer surface of themagnet 2. - Operation: placing the outer rings of the upper sides and lower sides of the plurality of
nickel grids 4 correspondingly on the twoadjacent arms 101 magnetized by themagnet 2. Thenickel grids 4 are fixed by the magnetic force of thearms 101 so as to achieve the batch immunolabeling operation of thenickel grids 4. Thenickel grids 4 can be prevented from carrying too much residual liquid in the process of transferring between different reaction liquids by the hydrophobic effect of the hydrophobic layer. - As shown in
FIG. 3 , a magnetic holder for immunoelectron microscopy grids with magnets located inside arms comprises aframe 1, themagnets 2 and ahydrophobic layer 3. Theframe 1 comprises at least twoaxisymmetric arms 101 and a connectingpart 102. The overall thickness of onearm 101 is not less than 0.1 mm. The interiors of the arms are hollow, and the exteriors of the arms are made of a solid material. Thearms 101 are connected by the connectingpart 102. The distance between the twoadjacent arms 101 is 2.6 to 3.3 mm. Themagnets 2 are magnet iron located in the hollow interiors of thearms 101. Thehydrophobic layer 3 adheres to the outer surface of theframe 1. - Operation: placing the outer rings of the upper sides and lower sides of the plurality of
nickel grids 4 correspondingly on the twoadjacent arms 101. Thenickel grids 4 are fixed by the magnetic force of the magnetic bodies inside thearms 101 so as to achieve the batch immunolabeling operation of thenickel grids 4. Thenickel grids 4 can be prevented from carrying too much residual liquid in the process of transferring between different reaction liquids by the hydrophobic effect of the hydrophobic layer.
Claims (14)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010015283.2 | 2020-01-07 | ||
CN202010015283.2A CN111122631A (en) | 2020-01-07 | 2020-01-07 | Magnetic suction type net carrying frame device |
Publications (2)
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US11041820B1 US11041820B1 (en) | 2021-06-22 |
US20210208088A1 true US20210208088A1 (en) | 2021-07-08 |
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Family Applications (1)
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US17/140,043 Active US11041820B1 (en) | 2020-01-07 | 2021-01-02 | Magnetic holder for immunoelectron microscopy grids |
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US (1) | US11041820B1 (en) |
CN (1) | CN111122631A (en) |
DE (1) | DE212020000728U1 (en) |
WO (1) | WO2021138936A1 (en) |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
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US20090325223A1 (en) * | 2008-06-24 | 2009-12-31 | Timothy Boyd | Magnetic immunohistochemical staining device and methods of use |
CN202757821U (en) * | 2012-07-11 | 2013-02-27 | 南京金斯瑞生物科技有限公司 | Reserving device for liquid on solid surface |
US20140169533A1 (en) * | 2012-12-19 | 2014-06-19 | Michael R. Razzano | System and method for capturing medical images |
CN103308671B (en) * | 2013-05-22 | 2016-02-03 | 北京康彻思坦生物技术有限公司 | A kind of detection film and detection system |
US9869648B2 (en) * | 2014-06-26 | 2018-01-16 | The Board Of Trustees Of The Leland Stanford Junior University | High density grids |
US10849442B2 (en) * | 2014-10-24 | 2020-12-01 | Leer, Inc. | Ice merchandiser with sensing capabilities |
CN105910875B (en) | 2016-05-27 | 2019-05-14 | 河南中镜科仪科技有限公司 | A kind of fixed device of load sample copper mesh |
CN207850957U (en) * | 2018-02-05 | 2018-09-11 | 南京医科大学 | A kind of fixing device of used in transmission electron microscope contained network |
-
2020
- 2020-01-07 CN CN202010015283.2A patent/CN111122631A/en active Pending
- 2020-01-17 DE DE212020000728.4U patent/DE212020000728U1/en active Active
- 2020-01-17 WO PCT/CN2020/072643 patent/WO2021138936A1/en active Application Filing
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2021
- 2021-01-02 US US17/140,043 patent/US11041820B1/en active Active
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Publication number | Publication date |
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CN111122631A (en) | 2020-05-08 |
DE212020000728U1 (en) | 2022-08-08 |
WO2021138936A1 (en) | 2021-07-15 |
US11041820B1 (en) | 2021-06-22 |
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