U.S. patent application number 17/140045 was filed with the patent office on 2021-07-08 for dual-arm clamping type holder for transmission electron microscopy grids and preparation method thereof.
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 Zheng Fan, Chunli Li, Hao Liang, Jingnan Liang, Yiwei Liu, Yuanming Luo, Shutao Sun, Weina Sun, Qian Wang, Xiaolan Zhang, Tong Zhao.
Application Number | 20210208090 17/140045 |
Document ID | / |
Family ID | 1000005666512 |
Filed Date | 2021-07-08 |
United States Patent
Application |
20210208090 |
Kind Code |
A1 |
Liang; Jingnan ; et
al. |
July 8, 2021 |
DUAL-ARM CLAMPING TYPE HOLDER FOR TRANSMISSION ELECTRON MICROSCOPY
GRIDS AND PREPARATION METHOD THEREOF
Abstract
The present invention relates to a dual-arm clamping type holder
for transmission electron microscopy grids and preparation method
thereof. The preparation method comprises: firstly manufacturing a
frame with two adjacent arms located on a same plane and have a
clamping structure by a hard material; then putting the frame in a
molten adhering liquid so that the frame is dipped with the
adhering liquid; and finally, taking out the frame dipped with the
adhering liquid, and waiting for the adhering liquid to solidify
into adhering layers along the arms. The dual-arm clamping type
holder manufactured by the method of the present invention
comprises a frame and adhering layers; and the adhering layers
adhere to the inner sides of the clamping structure between the two
adjacent arms of the frame.
Inventors: |
Liang; Jingnan; (Beijing,
CN) ; Liang; Hao; (Beijing, CN) ; Liu;
Yiwei; (Beijing, CN) ; Sun; Weina; (Beijing,
CN) ; Li; Chunli; (Beijing, CN) ; Luo;
Yuanming; (Beijing, CN) ; Fan; Zheng;
(Beijing, CN) ; Zhao; Tong; (Beijing, CN) ;
Sun; Shutao; (Beijing, CN) ; Wang; Qian;
(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: |
1000005666512 |
Appl. No.: |
17/140045 |
Filed: |
January 2, 2021 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 2223/309 20130101;
G01N 23/2202 20130101; G01N 23/2204 20130101 |
International
Class: |
G01N 23/2204 20060101
G01N023/2204; G01N 23/2202 20060101 G01N023/2202 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 7, 2020 |
CN |
202010014620.6 |
Claims
1. A preparation method for a dual-arm clamping type holder for
transmission electron microscopy grids, comprising: a.
manufacturing a frame from a hard material; wherein the frame has
at least two arms, and two adjacent arms are linked and located on
a same plane and have a clamping structure; b. putting the frame in
a molten adhering liquid, so that the frame is dipped with the
adhering liquid; c. taking out the frame dipped with the adhering
liquid, and waiting for the adhering liquid to solidify into
adhering layers.
2. The preparation method of claim 1, wherein step c further
comprises removing the adhering layers except on inner sides of two
adjacent arms of the frame.
3. A dual-arm clamping type holder for transmission electron
microscopy grids, comprising: a frame having at least two arms and
a connecting part, wherein two adjacent arms at two sides of the
connecting part are located on a same plane and have a clamping
structure; adhering layers adhered to inner sides of the clamping
structure between the two adjacent arms of the frame; and a
plurality of metal grids for transmission electron microscopy
supported by the frame, each metal grid comprising a metal ring and
a mesh surrounded by the metal ring and having a top surface, at
least one of the metal grids being disposed between the two
adjacent arms of the frame with the top surface of the metal grid
facing in a lengthwise direction of the two adjacent arms and with
the metal ring fixed to the adhering layers on the inner sides of
the two adjacent arms.
4. The dual-arm clamping type holder of claim 3, wherein the frame
is made of a hard material.
5. The dual-arm clamping type holder of claim 3, wherein the
adhering layers are made of a soft material.
6. The dual-arm clamping type holder of claim 3, wherein the
longitudinal section of the dual-arm clamping type holder is single
U-shaped.
7. The dual-arm clamping type holder of claim 3, wherein the
longitudinal section of the dual-arm clamping type holder is
parallel spiral type U-shaped.
8. The dual-arm clamping type holder of claim 3, wherein the
longitudinal section of the dual-arm clamping type holder is a
closed ring-shaped.
9. The dual-arm clamping type holder of claim 3 wherein the
adhering layers are adhered only to the inner sides of the two
adjacent arms.
10. The dual-arm clamping type holder of claim 3 wherein the two
adjacent arms extend parallel to each other.
11. The dual-arm clamping type holder of claim 3 wherein the
adhering layers are softer than the arms.
12. The dual-arm clamping type holder of claim 11 wherein the
adhering layers comprise wax.
13. A dual-arm holder for transmission electron microscopy grids
comprising: a frame comprising first and second arms each having a
lengthwise direction parallel to the lengthwise direction of the
other arm, and a first connecting part connected to a lengthwise
end of each arm, each arm having an inner side facing the other arm
and an outer side facing away from the other arm; an adhering layer
adhered to the inner side of each arm; and a plurality of metal
grids for electron microscopy supported by the frame, each metal
grid comprising a circular metal ring and a mesh which is
surrounded by the metal ring and has a top surface, at least one of
the metal grids being disposed between the first and second arms
with the top surface of the mesh facing in the lengthwise direction
of the arms and with the metal ring fixed to the adhering layer on
the inner side of each arm.
14. A dual-arm holder as claimed in claim 13 wherein a plurality of
the metal grids are disposed between the first and second arms with
the top surface of the mesh of each metal grid facing in the
lengthwise direction of the arms and with the metal ring of each
metal grid fixed to the adhering layer on the inner side of each
arm.
15. A holder as claimed in claim 13 wherein the adhering layer is
not adhered to the outer sides of the arms.
16. A holder as claimed in claim 13 wherein the adhering layer is
softer than the arms.
17. A holder as claimed in claim 16 wherein the adhering layer
comprises wax.
18. A holder as claimed in claim 13 wherein the arms and the first
connecting part form the shape of a single U.
19. A holder as claimed in claim 13 including a second connecting
part connected to the first and second arms at an opposite
lengthwise end of the arms from the first connecting part, wherein
the arms and the first and second connecting parts form an
elongated loop.
20. A holder as claimed in claim 13 further comprising a third arm
extending parallel to the first and second arms, and a second
connecting part connected between a lengthwise end of the second
arm and a lengthwise end of the third arm at an opposite lengthwise
end of the first and second arms from the first connecting part,
wherein the adhering layer is adhered to a surface of the third arm
opposing the second arm and a surface of the second arm opposing
the third arm, and at least one of the metal grids is disposed
between the second and third arms with the top surface of the mesh
of the metal grid facing in the lengthwise direction of the third
arm and with the metal ring of the metal grid fixed by the adhering
layer on the second and third arms.
Description
[0001] This application claims the priority of Chinese Patent
Application No. 202010014620.6, filed on Jan. 7, 2020, the entire
disclosure of which is incorporated herein by reference.
TECHNICAL FIELD
[0002] The invention relates to a holder for carrying grids with
biological ultrasections applied to observe biology ultrastructure
by transmission electron microscope, in particular to a holder for
carrying the grids with a high throughput in the sample staining
process.
BACKGROUND ART
[0003] A metal grid is a sample carrier used in a transmission
electron microscope. A copper grid is the most common metal grid.
The copper grid comprises two parts which are respectively a copper
ring with a rim width of about 0.2 mm and hollow grids inside the
copper ring. The thickness of the whole copper grid is about 18
micrometers, and there is an organic Formvar membrane on the
surface of the copper grid, which can effectively support section
of samples. The staining process is occurred on the copper grid.
Usually a pair of tweezers is needed for clamping the position of
outboard rim width around the copper grid during the staining
process. If the grid inner area of the copper grid is clamped by
the tweezers, the Formvar membrane adhering to the surfaces of the
grids may easily be damaged, and then cell sections loaded on the
copper grid may be damaged, which affects observation. As the
number of the grids with sections treated each time is large (4 to
20 grids with sections or even more), if each copper grid is
stained one by one, the staining times of the sections on the grids
are not uniform, and the sections are vulnerable to mechanical
damage due to repeated operations.
[0004] In order to achieve the unification of the staining time of
each grid with sections, 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 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 the arc-shaped opening or an
angle-shaped opening.
[0005] In the above-mentioned device, the main body adopts a
semi-closed structure, which causes the overall volume of the
device to be relatively large, ultimately leading to a need to use
more dye liquor when the device is used for batch staining. Besides
the opening part of the device, grooves or slits are formed between
the remaining part of the main body and the grids. When the device
is removed from the stain liquor after completing the staining
operation, some stain liquor in the grooves or slits will be taken
out, thereby increasing the stain liquid consumption. In addition,
the shielding formed by the semi-enclosed structure of the holder
increases the time of rinse and air-drying steps after staining.
Finally, the preparation of the above-mentioned device is highly
dependent on a mold. As the soft layers are inside the semi-closed
enclosing structure of the device, a complicated process is
required to achieve the adhesion and clamping efficiency of the
soft layers.
SUMMARY OF THE INVENTION
[0006] A main technical problem solved by the present invention is
to provide a preparation method for a dual-arm clamping type holder
for transmission electron microscopy grids which reduces dye loss,
improves the efficiency of cleaning and air-drying operations,
simplifies the preparation process, and can realize the batch
dyeing of copper grids, and a dual-arm clamping type holder for
transmission electron microscopy grids.
[0007] In order to solve the above technical problem, the present
invention provides a preparation method for a dual-arm clamping
type holder for transmission electron microscopy grids,
comprising:
[0008] a. manufacturing a frame by a hard material; wherein the
frame has at least two arms, and two adjacent arms are located on a
same plane and have a clamping structure;
[0009] b. putting the frame in a molten adhering liquid, so that
the frame is dipped with the adhering liquid; and
[0010] c. taking out the frame dipped with the adhering liquid, and
waiting for the adhering liquid to solidify into adhering
layers.
[0011] Further, a method of reducing the loss of the adhering
layers and improving the energy efficiency of the holder is
provided. Specifically, by removing the adhering layers except on
inner sides of two adjacent arms of the frame, not only can the
adhering layers be recycled to a great extent, but also the contact
area of the copper ring and wax layers can be reduced, so that the
pressure is stronger, and the copper grid is easily inserted into
and pulled out from the holder.
[0012] Further, a method of adjusting the thicknesses of the
adhering layers is also provided. Specially, the thickness of each
adhering layer decreases with the increase of liquid temperature,
increases with the increase of the number of dipping operations,
increases with the increase of dipping time, and increases with the
increase of the solidification degree of wax liquid after each
dipping operation except the last time.
[0013] A dual-arm clamping type holder for transmission electron
microscopy grids, comprising a frame and adhering layers, the
adhering layers adhere to inner sides of a clamping structure
between two adjacent arms of the frame.
[0014] Further, the frame is made of a hard material and provides a
basic surface for adhesion and molding of the adhering layers.
[0015] Further, the adhering layers are made of a soft material and
used for fixing and supporting the copper grid.
[0016] Further, a section of the dual-arm clamping type holder is
preferably single U-shaped.
[0017] Further, a section of the dual-arm clamping type holder is
preferably parallel spiral type U-shaped.
[0018] Further, a section of the dual-arm clamping type holder is
preferably closed ring-shaped.
[0019] In conclusion, the present invention has the following
beneficial effects:
[0020] by eliminating the redundant frame of the holder, the space
occupied by the holder is reduced. Further the amount of dye liquor
required for staining is reduced. At the same time, the open
structure adopted by the present invention effectively avoids the
formation of grooves or slits between the holder and the copper
grid, so that the amount of the dye liquor carried by the holder is
close to zero. In addition, by adopting the open structure, the
efficiency of a cleaning step and an air-drying step after dyeing
is also improved. Furthermore, the preparation method of the device
is simpler, and almost zero dependence on a mold can be realized
when the frame is manufactured. Improvement in clamping efficiency
of the adhering layers can be quickly realized by convenient
technologies such as directly cutting off the peripheral excess wax
layers along the wall of the frame.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] 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:
[0022] FIG. 1 shows a single U-shaped holder before efficiency
improvement and a cross section of the holder.
[0023] FIG. 2 shows a single U-shaped holder after efficiency
improvement and a cross-section of the holder.
[0024] FIG. 3 shows a single U-shaped holder loading a copper
grid.
[0025] FIG. 4 shows a parallel spiral type U-shaped holder loading
a copper grid.
[0026] FIG. 5 shows a closed ring-shaped holder loading a copper
grid.
[0027] FIG. 6 shows batch staining of a copper grid.
DETAILED DESCRIPTION OF THE INVENTION
[0028] 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
[0029] This example provides a preparation method for a dual-arm
clamping type holder for transmission electron microscopy grids,
which specially comprises the following steps:
[0030] step 1. manufacturing a frame;
[0031] winding a hard material (such as iron wires, steel wires,
glass and plastics) into the frame with at least two arms, wherein
two adjacent arms are located on a same plane and have a clamping
structure, and the distance between the two arms is 3-4.5 mm. The
frame is single U-shaped, parallelly spiral type U-shaped or closed
ring-shaped so as to provide basic surfaces for adhesion and
molding of wax layers.
[0032] step 2. adhering the wax layers;
[0033] a. putting a solid wax (food grade beeswax or dental wax)
into a container for heating;
[0034] b. after heating the solid wax to a completely molten state,
putting the frame into the wax liquid for dipping the frame in the
wax liquid;
[0035] Further, the thicknesses of the wax layers of the frame can
be adjusted by controlling the time and number of times of the
frame dipped in the wax liquid, the temperature of the wax liquid,
and the solidification degree of the wax liquid after each dipping
except the last time.
[0036] As shown in the table 1, under the condition that the single
dipping time, the number of times of dipping and the solidification
degree of the wax liquid after each dipping except the last time
are the same, the thicknesses of the wax layers are inversely
proportional to the temperature of the wax liquid. Under the
condition that the temperature of the wax liquid, the single
dipping time, and the solidification degree of the wax liquid after
each dipping except the last time are the same, the thicknesses of
the wax layers are directly proportional to the number of times of
dipping. And under the condition that the temperature of the wax
liquid, the single dipping time and the number of times of dipping
are the same, the thicknesses of the wax layers are directly
proportional to the solidification degree of the wax liquid after
each dipping except the last time.
TABLE-US-00001 TABLE 1 Thickness of wax layer under different
conditions Wax layer thickness (mm) Dipping Dipping 1s .times. 2 1s
.times.2 number number of times of times (Dipping (Dipping for for
the the second second time time after the Wax after the wax liquid
liquid wax liquid dipped at dipping Dipping Dipping dipped at the
first Wax liquid mode 1s .times. 1 2s .times. 1 the first time is
temperature ( error number number time is half completely (DEG C)
range) of times of times solidified) solidified) 80 Horizonal 0.215
0.23 0.30 0.40 (.+-.10%) Vertical 0.27 0.35 0.41 0.51 (.+-.25%) 75
Horizontal 0.235 0.25 0.50 0.64 (.+-.10%) Vertical 0.3 0.54 0.53
0.68 (.+-.25%) 70 Horizontal 0.375 0.45 0.55 0.71 (.+-.10%)
vertical 0.425 0.61 0.69 0.79 (.+-.25%)
[0037] c. taking the frame out of the wax liquid, and waiting for
the wax liquid adhering to the frame to naturally solidify.
[0038] step 3. reducing consumption and improving efficiency;
[0039] As shown in FIGS. 1 and 2, taking the horizontal plane of
the frame as the reference, the wax layers adhered to the inner
sides of the clamping structure between two adjacent arms of the
frame are retained, and the wax layers adhered to the front and
back of the frame outside are cut off, thus, the contact area when
the copper grid is inserted into the wax layers is reduced, the
pressure of the contact points between the grid and the wax layers
is increased, the insertion and extraction of the copper grid are
facilitated, and the deformation of the copper grid is reduced; and
the wax layers adhering to the outer edges of the two arms of the
frame are cut off so as to reduce wax consumption and recycle
wax.
[0040] The ideal working thicknesses of the wax layers on the inner
sides of the two adjacent arms of the holder are 0.5-1 mm; and the
ideal gap between the wax layers is 1-2.6 mm. When the wax layers
adsorbed by the entire frame are too thick or the wax layers
adhering to the lower part of the frame is too thick (after the
frame is taken out of the wax liquid, the wax liquid adhered to the
frame flows downward under the action of gravity, and thus, the wax
layers adhered to the frame are thin on the upper part and thick on
the lower part), the extra thick parts of the wax layers adhered to
the two adjacent arms of the holder can be cut off according to
actual needs so as to improve the efficiency or utilization rate of
the holder.
Example 2
[0041] FIG. 3 shows a single U-shaped holder loading a copper grid,
which is manufactured by the method of the present application. The
device comprises a single U-shaped frame 1 and adhering layers 2.
The two arms included in the frame 1 provide basic surfaces for
adhesion and molding of the adhering layers 2. The adhering layers
2 are used for fixing and supporting the copper grid 3. The copper
ring part of the copper grid 3 can be inserted into and fixed to
the inner sides of the adhering layers 2, and are arranged in
sequence along the long axes of the arms. Specially, the single
U-shaped frame 1 is made of a hard material (such as iron wires,
steel wires, glass, plastics). The adhering layers 2 are made of
wax layers (food grade beeswax or dental wax), and adhere to the
inner sides of the U-shaped structure of the single U-shaped frame
1. The distance between the inner sides of the two arms of the
single U-shaped frame 1 is 3-4.5 mm, and the distance between the
inner sides of the adhering layers 2 is 1-2.6 mm. The copper grid 3
is a copper ring with an outer diameter of 3 mm, an inner diameter
of 2.6 mm, and a rim width of about 0.2 mm. There are hollow grids
ranging from 50 to 400 meshes within the ring inner area. The
thickness of the entire copper grid 3 is about 18 micrometers. And
there is an organic Formvar membrane on the front of the copper
grid 3, which can effectively support section samples.
[0042] FIG. 4 shows a parallelly spiral type U-shaped holder
loading a copper grid, which is manufactured by the method of the
present application. The holder comprises a parallelly spiral type
U-shaped frame 1 and adhering layers 2. The two adjacent arms
included in the frame 1 provide the basic surfaces for adhesion and
molding of the adhering layers 2. The adhering layers 2 are used
for fixing and supporting the copper grid 3. The copper ring parts
of the copper grid 3 can be inserted into and fixed to the inner
sides of the adhering layers 2, and are arranged in sequence along
the long axes of the arms. Specially, the parallelly spiral type
U-shaped frame 1 is made of a hard materials (such as iron wires,
steel wires, glass, plastics). The adhering layers 2 are made of
wax layers (food grade beeswax or dental wax), and adhere to the
inner sides of the U-shaped structure of the parallelly spiral type
U-shaped frame 1. The distance between the inner sides of the two
adjacent arms of the parallelly spiral type U-shaped frame 1 is
3-4.5 mm, and the distance between the inner sides of the adhering
layers 2 is 1-2.6 mm. The copper grid 3 is a copper ring with an
outer diameter of 3 mm, an inner diameter of 2.6 mm, and a rim
width of about 0.2 mm. There are hollow grids ranging from 50 to
400 meshes within the ring inner area. The thickness of the entire
copper grid 3 is about 18 micrometers. And there is an organic
Formvar membrane on the front of the copper grid 3, which can
effectively support section samples.
[0043] FIG. 5 shows a closed ring-shaped holder loading a copper
grid, which is manufactured by the method of the present
application. The holder comprises a closed ring-shaped frame 1 and
adhering layers 2. The two arms of the frame 1 provide basic
surfaces for adhesion of the adhering layers 2. The adhering layers
2 are used for fixing and supporting the copper grid 3. The copper
ring parts of the copper grid 3 can be inserted into and fixed to
the inner sides of the adhering layers 2, and are arranged in
sequence along the long axes of the arms. Specially, the closed
ring-shaped frame 1 is made of a hard material (such as iron wires,
steel wires, glass). The adhering layers 2 are made of wax layers
(food grade beeswax or dental wax), and adhere to the inner sides
of the two arms of the closed ring-shaped frame 1. The distance
between the inner sides of the two arms of the closed ring-shaped
frame 1 is 3-4.5 mm, and the distance between the inner sides of
the adhering layers 2 is 1-2.6 mm. The copper grid 3 is a copper
ring with an outer diameter of 3 mm, an inner diameter of 2.6 mm,
and a rim width of about 0.2 mm. There are hollow grids ranging
from 50 to 400 meshes within the ring inner area. The thickness of
the entire copper grid 3 is about 18 micrometers. And there is an
organic Formvar membrane on the front of the copper grid 3, which
can effectively support section samples.
[0044] FIG. 6 shows batch staining of the copper grids with
sections, which is performed according to the present invention.
The distance between the adjacent copper grids fixed to the
adhering layers is greater than 2.5 mm to reduce the carry-over
amount of staining liquor 4. The part of the holder loading the
copper grids 3 is dipped under the liquid surface of the staining
liquor 4 so as to achieve the batch staining of the copper
grids.
* * * * *