U.S. patent application number 17/140043 was filed with the patent office on 2021-07-08 for magnetic holder for immunoelectron microscopy grids.
This patent application is currently assigned to Institute of Microbiology, Chinese Academy of Sciences. The applicant listed for this patent is Institute of Microbiology, Chinese Academy of Sciences. Invention is credited to Xing Dai, Zheng Fan, Zhousinuo Jiang, Chunli Li, Hao Liang, Jingnan Liang, Yiwei Liu, Yuanming Luo, Shutao Sun, Qian Wang, Xiaolan Zhang, Tong Zhao.
Application Number | 20210208088 17/140043 |
Document ID | / |
Family ID | 1000005666511 |
Filed Date | 2021-07-08 |
United States Patent
Application |
20210208088 |
Kind Code |
A1 |
Liang; Jingnan ; et
al. |
July 8, 2021 |
MAGNETIC HOLDER FOR IMMUNOELECTRON MICROSCOPY GRIDS
Abstract
The present invention relates to a magnetic holder for
immunoelectron microscopy grids. The holder comprises a frame, a
magnet and a hydrophobic layer. The device can use a magnetic force
to simultaneously attach the outer rings of nickel grids to the
frame, so that a batch operation (such as rinsing, immunolabeling
and dyeing) of the nickel grids can be realized. In addition, due
to the hydrophobic effect of the hydrophobic layer, the holder can
reduce the amount of the liquid carried by the nickel grids in the
process of continuously transferring the nickel grids between
different types of liquids to almost zero. Compared with the prior
art, the magnetic holder effectively reduces the probability of
cross-contamination between reagents.
Inventors: |
Liang; Jingnan; (Beijing,
CN) ; Liu; Yiwei; (Beijing, CN) ; Wang;
Qian; (Beijing, CN) ; Jiang; Zhousinuo;
(Beijing, CN) ; Dai; Xing; (Beijing, CN) ;
Liang; Hao; (Beijing, CN) ; Luo; Yuanming;
(Beijing, CN) ; Li; Chunli; (Beijing, CN) ;
Fan; Zheng; (Beijing, CN) ; Zhao; Tong;
(Beijing, CN) ; Sun; Shutao; (Beijing, CN)
; Zhang; Xiaolan; (Beijing, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Institute of Microbiology, Chinese Academy of Sciences |
Beijing |
|
CN |
|
|
Assignee: |
Institute of Microbiology, Chinese
Academy of Sciences
Beijing
CN
|
Family ID: |
1000005666511 |
Appl. No.: |
17/140043 |
Filed: |
January 2, 2021 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 2223/612 20130101;
G01N 2223/309 20130101; G01N 23/2202 20130101; G01N 23/2204
20130101; G01N 33/532 20130101 |
International
Class: |
G01N 23/2202 20060101
G01N023/2202; G01N 23/2204 20060101 G01N023/2204; G01N 33/532
20060101 G01N033/532 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 7, 2020 |
CN |
202010015283.2 |
Claims
1. A magnetic holder for nickel grids [] for use in immunoelectron
microscopy, comprising a frame, a first magnet a hydrophobic layer
adhered to an outer surface of the holder, and a plurality of
nickel grids fixed to a surface of the frame by the first magnet,
each nickel grid having a nickel ring and a mesh grid disposed
inside the nickel ring.
2-18. (canceled)
19. A holder as claimed in claim 1 wherein: each nickel ring has an
inner diameter and an outer diameter: the frame comprises first and
second arms opposing each other and spaced from each other by a gap
which is smaller than the outer diameter of the nickel rings and
which is greater than or equal to the inner diameter of the nickel
rings; the first magnet contacts the frame and generates a magnetic
force through the frame which fixes the nickels grids to the frame
with the nickel ring of each nickel grid spanning the gap between
the two arms of the frame.
20. A holder as claimed in claim 19 wherein each arm has a flat
surface adjoining the gap for supporting the nickel rings of the
nickel grids fixed to the frame by the first magnet.
21. A holder as claimed in claim 19 wherein the hydrophobic layer
adheres to the first magnet.
22. A holder as claimed in claim 19 wherein: each arm of the frame
has a plurality of arc-shaped grooves formed along the gap in an
upper surface of the arm and spaced from each other in a lengthwise
direction of the arm, each groove having an opening which faces the
opposing arm, each groove being axisymmetric with respect to one of
the grooves in the opposing arm, each groove accommodating a
portion of the periphery of the nickel ring of one of the nickel
grads spanning the gap between the arms.
23. A holder as claimed in claim 22 wherein each groove extends
partway through a depth of the arm in which the groove is
formed.
24. A holder as claimed in claim 22 wherein the first magnet is
detachably arranged below and in contact with the arms of the
frame.
25. A holder as claimed in claim 19 wherein: the first magnet is
located in the gap between the two arms and connects and magnetizes
the arms and generates a magnetic force which fixes the nickel
grids to the frame with the nickel grids spanning the gap.
26. A holder as claimed in claim 25 wherein each arm has a length
and a width, and the first magnet has a length which is smaller
than the length of either arm and a width measured in a widthwise
direction of the frame which is smaller than the width of either
arm.
27. A holder as claimed in claim 25 wherein each arm is made of a
magnetizable material.
28. A holder as claimed in claim 19 further comprising a second
magnet and a connecting member which extends between and connects
the first and second arms, wherein the first magnet is disposed in
a hollow interior of the first arm and the second magnet is
disposed in a hollow interior of the second arm, and the first and
second magnets generate magnetic forces which fix the nickel grids
to the frame with the nickel grids spanning the gap between the
arms.
29. A holder as claimed in claim 28 wherein the first arm surrounds
a periphery of the first magnet and the second arm surrounds a
periphery of the second magnet.
30. A holder as claimed in claim 28 wherein the connecting member
is connected to a lengthwise end of each arm.
Description
[0001] 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.
TECHNICAL FIELD
[0002] 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.
BACKGROUND ART
[0003] 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.
[0004] 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.
[0005] 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.
SUMMARY OF THE INVENTION
[0006] 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.
[0007] 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
[0008] 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.
[0009] 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.
[0010] 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.
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] 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:
[0012] FIG. 1 is a schematic diagram of a magnetic holder with a
frame located at one side of a magnet.
[0013] FIG. 2 is a schematic diagram of a magnetic holder with a
magnet located between two adjacent arms.
[0014] FIG. 3 is a schematic diagram of a magnetic holder with
magnets located inside arms, showing a A-A' cross section.
DETAILED DESCRIPTION OF THE INVENTION
[0015] 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.
EXAMPLE 1
[0016] As shown in FIG. 1, 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.
[0017] Operation: separating the frame 1 and the magnet 2, placing
the outer rings of the upper sides and lower sides of the plurality
of nickel grids 4 respectively in the symmetrical arc-shaped
grooves of the arms 101, recombining the frame 1 loaded the nickel
grids 4 and the magnet 2, and carrying out the immunolabeling
operation. After the immunolabeling is completed, separating the
frame 1 and the magnet 2 again. Finally taking out the nickel grids
4. The nickel grids 4 are fixed by the magnetic force of the magnet
2 under the frame 1 to achieve the batch staining for the nickel
grids 4. The nickel 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 the frame 1 from the magnet 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.
EXAMPLE 2
[0018] As shown in FIG. 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.
[0019] Operation: placing the outer rings of the upper sides and
lower sides of the plurality of nickel grids 4 correspondingly on
the two adjacent arms 101 magnetized by the magnet 2. The nickel
grids 4 are fixed by the magnetic force of the arms 101 so as to
achieve the batch immunolabeling operation of the nickel grids 4.
The nickel 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.
EXAMPLE 3
[0020] As shown in FIG. 3, 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.
[0021] Operation: placing the outer rings of the upper sides and
lower sides of the plurality of nickel grids 4 correspondingly on
the two adjacent arms 101. The nickel grids 4 are fixed by the
magnetic force of the magnetic bodies inside the arms 101 so as to
achieve the batch immunolabeling operation of the nickel grids 4.
The nickel 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.
* * * * *