CN210181050U

CN210181050U - Anti-collision temperature-control antibody incubation box

Info

Publication number: CN210181050U
Application number: CN201920855483.1U
Authority: CN
Inventors: Zhizi Tong; 童智子
Original assignee: Southwest University
Current assignee: Southwest University
Priority date: 2019-06-06
Filing date: 2019-06-06
Publication date: 2020-03-24
Anticipated expiration: 2029-06-06

Abstract

The utility model discloses a scour protection hits accuse temperature antibody and hatches box relates to biological experiment equipment technical field, has solved that current antibody is hatched the box and is slightly rocked or place not just can make an anti (liquid) from the epimembranal outflow to lead to the membrane to become dry to cause the problem of influence to final experimental result. The key points of the technical scheme are as follows: the box body is composed of a first incubation grid, a second incubation grid, a third incubation grid and a fourth incubation grid, wherein the bottom surface and the top surface of the box body are positioned on the same plane and are different in size; the box body is provided with a box cover for covering the top opening of the box body; the inner wall of the box body and the bottom surface of the box cover are provided with super-hydrophobic nano coatings. This hatch box can prevent among the test process box body when bumping that the antibody diluent from splashing to hatch the box wall on the messenger and place the membrane in the box body bottom dry to can guarantee the effect of anti hatching, and can reduce the waste of time, space, resource and the cost among the test process effectively.

Description

Anti-collision temperature-control antibody incubation box

Technical Field

The utility model relates to a biological assay equipment technical field, more specifically say that it relates to scour protection hits accuse temperature antibody and hatches box.

Background

Immunology is the key research field in the contemporary medical and life science fields, and provides a key idea for disease prevention, treatment and research and development of new drugs. Immunoblotting (Western Blotting) is an international and common method for detecting proteins in immunological studies. However, many factors in the practical operation of this experimental technique can lead to the final result being not ideal. The incubation process of the primary antibody is one of the key steps in the experimental method.

At present, the main structure of the existing antibody incubation box for primary antibody incubation comprises a cuboid box body made of common materials and a box cover connected with the top end of the box body.

In the prior art, when an antibody incubation box is used, a PVDF membrane in an experimental material is placed in the antibody incubation box, and corresponding antibody diluent is dripped on the PVDF membrane according to the protein to be detected; then, putting the antibody incubation box into a refrigeration box to carry out an antibody incubation experiment; the existing antibody incubation box is slightly shaken or placed unevenly, so that the antibody (liquid) flows out of the membrane once, the membrane is dried, and the final result is possibly influenced greatly. In addition, the inner wall of the existing antibody incubation box is inconvenient to clean after the test is carried out.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a scour protection hits accuse temperature antibody and hatches box, can prevent that the antibody diluent from splashing to the box wall of hatching when box body is collided among the test process and make the membrane of placing in the box body bottom dry to can guarantee the effect of anti hatching, and can reduce the waste of time, resource and the cost among the test process effectively.

The above technical purpose of the present invention can be achieved by the following technical solutions: the anti-collision temperature-control antibody incubation box comprises a box body, wherein the box body is composed of a first incubation grid, a second incubation grid, a third incubation grid and a fourth incubation grid, the bottom surface and the top surface of the box body are positioned on the same plane, and the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are different in size; the box body is provided with a box cover for covering the top opening of the box body; the inner wall of the box body and the bottom surface of the box cover are provided with super-hydrophobic nano coatings.

By adopting the technical scheme, in the process of performing primary antibody incubation by using the antibody incubation box, firstly, the washed PVDF membrane is clamped by using tweezers according to the width of a strip of a protein immunoblot, the PVDF membrane is selectively placed in a first incubation lattice, a second incubation lattice, a third incubation lattice or a fourth incubation lattice, the back surface of the PVDF membrane is attached to the bottom end of the inner wall of the box body, and then the PVDF membrane is dropwise added with corresponding types of antibody diluents according to the protein to be detected; through the first incubation grids, the second incubation grids, the third incubation grids and the fourth incubation grids with different sizes, the antibody incubation of strips with different widths is facilitated; through the box cover, the covering of the openings at the top ends of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid and the super-hydrophobic effect of the super-hydrophobic nano coating are facilitated, so that the antibodies in the box body are prevented from volatilizing in a large amount; through the hydrophobic effect of the super-hydrophobic nano coating and a certain inclination of the side wall of the box body, the antibody is prevented from splashing out of the surface of the PVDF membrane due to collision in the experimental process, and the antibody cannot return to the surface of the PVDF membrane, so that the membrane is dried to cause experimental failure; meanwhile, the box body is convenient to clean through the super-hydrophobic nano layer, and pollution to subsequent experiments is avoided, so that water resources are saved to a certain extent, and an environment-friendly effect is achieved; through the first anti-collision temperature-control antibody incubation box that incubate check, second incubation check, third incubation check, fourth incubation check, lid and super-hydrophobic nano layer constitute, the antibody diluent splashes to the incubation box wall when can prevent among the test process box body from colliding and makes the membrane of placing in the box body bottom dry to can guarantee the result of anti-incubation, and can reduce the waste of time, resource and the expenditure among the test process effectively.

The utility model discloses further set up to: the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are inverted truncated pyramid bodies; the end of the first incubation grid with the open top is connected with the end of the second incubation grid with the open top, the end of the second incubation grid with the top far from the first incubation grid is connected with the end of the third incubation grid with the open top, and the end of the third incubation grid with the top far from the second incubation grid is connected with the end of the fourth incubation grid with the open top.

Through adopting above-mentioned technical scheme, when box body collided in the test process, had certain gradient and scribbled the first check of hatching, the check is hatched to the second, the check is hatched to the third and the check is hatched to the fourth of super hydrophobic coating through the lateral wall, is convenient for make the antibody diluent who splashes on the box body inner wall slide back box body inner wall bottom.

The utility model discloses further set up to: the length of the top end of the box body is 13.0cm, and the width of the top end of the box body is 9.4 cm; the side surface of the box body is in an inverted trapezoid shape, and the height of the box body is 1.7 cm; the widths of openings at the top ends of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are respectively 4.0cm, 3.5cm, 3.0cm and 2.5 cm; the bottom surfaces of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are 8.4cm in length; the widths of the bottom surfaces of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are 3.0cm, 2.5cm, 2.0cm and 1.5cm respectively; the distance between the bottom surfaces of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid is 1.0 cm.

By adopting the technical scheme, the widths of the openings at the top ends of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are respectively 3.0cm, 2.5cm, 2.0cm and 1.5cm, so that PVDF films with different sizes can be conveniently placed into the corresponding incubation grids according to requirements.

The utility model discloses further set up to: the cross section of the box cover is rectangular, the length of the box cover is 10.1cm, and the width of the box cover is 9.5 cm; the bottom surface of the box cover is provided with a raised interface which is matched with openings at the top ends of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid, and the raised interface is rectangular.

By adopting the technical scheme, the top openings of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are conveniently covered by the box cover with the length of 10.1cm and the width of 9.5 cm; through protruding interface, be convenient for lid and first incubation check, second incubation check, third incubation check and the connection of fourth incubation check top are fixed.

The utility model discloses further set up to: the side walls of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid are respectively provided with a refrigerating sheet and a film type temperature sensor which are used for controlling the temperature in the box body, and the temperature control range of the refrigerating sheet is 4-37 ℃; the box body is provided with a singlechip which is connected with the refrigerating sheet and the membrane type temperature sensor; and the side wall of the box body is provided with a solar cell.

By adopting the technical scheme, in the process of carrying out the antibody incubation test, the temperature information of the outer walls of the first incubation grid, the second incubation grid, the third incubation grid and the fourth incubation grid can be sensed conveniently in real time through the film type temperature sensor, and the sensed temperature information is transmitted to the single chip microcomputer; the single chip microcomputer receives the temperature information transmitted by the membrane type temperature sensor and then processes and analyzes the temperature information, the temperature information is compared with an antibody incubation temperature value implanted into the single chip microcomputer in advance, if the temperature information is higher than the antibody incubation temperature value, the single chip microcomputer controls the refrigerating sheet to work, so that the first incubation lattice, the second incubation lattice, the third incubation lattice and the fourth incubation lattice are cooled, when the temperature information sensed by the membrane type temperature sensor is located at the antibody incubation temperature value implanted into the single chip microcomputer in advance, the single chip microcomputer controls the refrigerating sheet to stop cooling work, so that the temperature of antibody incubation in the box body can be kept in a stable range, and therefore the test result of antibody incubation can be conveniently guaranteed; meanwhile, a temperature control system consisting of the refrigerating sheet, the film type temperature sensor, the solar cell sheet and the single chip microcomputer avoids the situation that the incubation box is easily collided by opening and closing the refrigerator door or the incubator door for multiple times after the incubation box is placed in the refrigerator or the incubator in the traditional antibody incubation process, and the waste of space resources is reduced; meanwhile, the solar cell is utilized, the influence of electricity interruption on the incubation temperature of the antibody in the incubation box in the traditional antibody incubation process can be avoided, and the effect of saving energy is achieved.

To sum up, the utility model discloses following beneficial effect has: through the first incubation grids, the second incubation grids, the third incubation grids and the fourth incubation grids with different sizes, the antibody incubation of strips with different widths is facilitated; through the box cover, the first incubation grids, the second incubation grids, the third incubation grids and the fourth incubation grids are conveniently covered by openings at the top ends, so that the antibodies in the box body are conveniently prevented from volatilizing in large quantity; the super-hydrophobic nano coating is convenient for preventing the antibody from splashing out of the surface of the PVDF membrane to cause experiment failure due to collision in the experiment process; meanwhile, the box body is convenient to clean through the super-hydrophobic nano layer, and pollution to subsequent experiments is avoided, so that water resources are saved to a certain extent, and an environment-friendly effect is achieved; through the first scour protection that incubate check, second are incubated check, the check is incubated to the third, the check is incubated to the fourth, the case lid is incubated with accuse temperature antibody and is hatched the box that the super hydrophobic nanometer layer constitutes, can prevent that the antibody diluent from splashing when the box body is collided in the experimentation and making the membrane of placing in the box body bottom dry on incubating the box wall, thereby can guarantee an anti effect of incubating, and can reduce time and the waste of expenditure in the experimentation effectively.

Drawings

Fig. 1 is a schematic structural diagram of a box body in an embodiment of the present invention;

FIG. 2 is a schematic view of the structure of FIG. 1 taken along line A-A;

FIG. 3 is a side view of a cartridge body in an embodiment of the invention;

fig. 4 is a schematic structural view of the cover in the embodiment of the present invention;

fig. 5 is a front view of the box body in the embodiment of the present invention.

In the figure: 1. a box body; 2. a first incubation compartment; 3. a second incubation compartment; 4. a third incubation compartment; 5. a fourth incubation compartment; 6. a box cover; 7. a superhydrophobic nanocoating; 8. a refrigeration plate; 9. a film-type temperature sensor; 10. a single chip microcomputer; 11. a raised interface; 12. provided is a solar cell.

Detailed Description

The present invention will be described in further detail with reference to fig. 1 to 5.

Example (b): as shown in fig. 1, 2, 3, 4 and 5, the anti-collision temperature-controlled antibody incubation case includes a case body 1, and the case body 1 is composed of a first incubation lattice 2, a second incubation lattice 3, a third incubation lattice 4 and a fourth incubation lattice 5, which have different sizes and whose bottom surfaces and top surfaces are located on the same plane. The box body 1 is provided with a box cover 6 for covering the top end opening of the box body 1. The inner wall of the box body 1 and the bottom surface of the box cover 6 are coated with a super-hydrophobic nano coating 7.

In the embodiment, the superhydrophobic nano-coating 7 is made of a superhydrophobic nano-material, and the surface of the superhydrophobic nano-material has lower free energy than that of a common material, so that the hydrophobicity of the solid surface can be improved. Meanwhile, the super-hydrophobic nano material has high melting point and stable property, and the lowest surface free energy can be obtained on a flat solid surface through compact and ordered arrangement. The box body is made of polypropylene (PP) material. In the process of primary antibody incubation by using the antibody incubation box, firstly, the washed PVDF membrane is clamped by forceps according to the width of a strip of a protein immunoblot, is selectively placed into the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 or the fourth incubation grid 5, the back of the PVDF membrane is attached to the bottom end of the inner wall of the box body 1, and then the antibody diluent of the corresponding type is dripped on the PVDF membrane according to the protein to be detected. The incubation of antibodies of different width bands is facilitated by the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 and the fourth incubation grid 5, which are different in size. Through lid 6, be convenient for first hatch check 2, the second is hatched check 3, the third is hatched check 4 and the fourth is hatched 5 open-top covers of check to be convenient for guarantee that the antibody in the box body 1 can not volatilize in a large number. The super-hydrophobic nano coating 7 is convenient for preventing the antibody from splashing out of the surface of the PVDF membrane to cause experiment failure due to collision in the experiment process. Meanwhile, through the super-hydrophobic nano layer, the box body 1 is convenient to clean, and pollution to subsequent experiments can be avoided, so that water resources are saved to a certain extent, and the environment-friendly effect is achieved. Through first hatching check 2, second hatching check 3, the third is hatched check 4, the check 5 is hatched to the fourth, the box is hatched to the scour protection that check 6 and super hydrophobic nanometer layer constitute and is hit accuse temperature antibody, can prevent that the antibody diluent from splashing when box body 1 is collided among the experimentation and making the membrane of placing in 1 bottom of box body dry on hatching the box wall, thereby can guarantee the result of anti hatching, and can reduce time and the expenditure among the experimentation effectively.

The first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4, and the fourth incubation lattice 5 are inverted truncated pyramid bodies. The end of the first incubation lattice 2 having the open top is connected to the end of the second incubation lattice 3 having the open top, the end of the second incubation lattice 3 having the open top away from the first incubation lattice 2 is connected to the end of the third incubation lattice 4 having the open top, and the end of the third incubation lattice 4 having the open top away from the second incubation lattice 3 is connected to the end of the fourth incubation lattice 5 having the open top.

In this embodiment, when the box body 1 is collided during the test, the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4 and the fourth incubation lattice 5 which are inverted truncated pyramid bodies are used, so that the side walls and the bottom surfaces of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4 and the fourth incubation lattice 5 have a certain inclination angle and are provided with the superhydrophobic nano-coating, and thus the antibody diluent splashed on the inner wall of the box body 1 can slide back to the bottom end of the inner wall of the box body 1.

The top end of the box body 1 is 13.0cm in length and 9.4cm in width. The side of box body 1 is for falling trapezoidal, and the height of box body 1 is 1.7 cm. The widths of the top openings of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4, and the fourth incubation lattice 5 are 4.0cm, 3.5cm, 3.0cm, and 2.5cm, respectively. The length of the bottom surface of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4, and the fourth incubation lattice 5 is 8.4 cm. The widths of the bottom surfaces of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4, and the fourth incubation lattice 5 are 3.0cm, 2.5cm, 2.0cm, and 1.5cm, respectively. The distance between the bottom surfaces of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4, and the fourth incubation lattice 5 is 1.0 cm.

In this embodiment, the widths of the top openings of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4 and the fourth incubation lattice 5 are 3.0cm, 2.5cm, 2.0cm and 1.5cm, respectively, so that PVDF membranes of different sizes can be placed in the corresponding incubation lattices as needed.

The cross section of the box cover 6 is rectangular, the length of the box cover 6 is 10.1cm, and the width of the box cover 6 is 9.5 cm. The bottom surface of the box cover 6 is provided with a raised interface 11 which is matched with the top openings of the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 and the fourth incubation grid 5, and the raised interface 11 is in a rectangular frame shape.

In this embodiment, the top ends of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4 and the fourth incubation lattice 5 are covered by the box cover 6 having a length of 10.1cm and a width of 9.5 cm. The box cover 6 is conveniently connected and fixed with the top ends of the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 and the fourth incubation grid 5 through the convex interfaces 11.

The side walls of the first incubation grids 2, the second incubation grids 3, the third incubation grids 4 and the fourth incubation grids 5 are respectively provided with a refrigerating sheet 8 and a film type temperature sensor 9 which are used for controlling the temperature in the box body 1, and the temperature control range of the refrigerating sheet 8 is 4-37 ℃. The box body 1 is provided with a singlechip 10 which is connected with a refrigerating sheet 8 and a film type temperature sensor 9. The side wall of the box body 1 is provided with a solar cell 12.

In this embodiment, in the process of performing the antibody incubation test, the temperature information of the outer walls of the first incubation lattice 2, the second incubation lattice 3, the third incubation lattice 4 and the fourth incubation lattice 5 is sensed in real time through the film-type temperature sensor 9, and the sensed temperature information is transmitted to the single chip microcomputer 10. Singlechip 10 handles this temperature information after receiving the temperature information of diaphragm type temperature sensor 9 transmission and analysis, and with this temperature information and the antibody incubation temperature value of implanting in singlechip 10 in advance contrast, if this temperature information is higher than the antibody incubation temperature value, singlechip 10 control refrigeration piece 8 works, thereby to first hatching check 2, check 3 is hatched to the second, check 4 is hatched to the third and check 5 is hatched to the fourth, when the temperature information that senses as diaphragm type temperature sensor 9 is located the antibody incubation temperature value of implanting in singlechip 10 in advance, singlechip 10 control refrigeration piece 8 stops cooling work, thereby make the temperature that the antibody in the box body 1 was hatched can keep in stable range, thereby be convenient for guarantee the experimental result that the antibody was hatched. Meanwhile, a temperature control system consisting of the refrigerating plate 8, the membrane type temperature sensor 9, the solar cell plate 12 and the single chip microcomputer 10 is utilized, so that the collision of the incubation box caused by opening and closing the refrigerator door or the incubator door for multiple times after the incubation box is placed in a refrigerator or an incubator in the traditional antibody incubation process is avoided, and the waste of space resources is reduced; meanwhile, the solar cell 12 can avoid the influence of electricity interruption on the incubation temperature of the antibody in the incubation box in the traditional antibody incubation process, and the effect of saving energy is achieved.

The working principle is as follows: in the process of primary antibody incubation by using the antibody incubation box, firstly, the washed PVDF membrane is clamped by forceps according to the width of a strip of a protein immunoblot, is selectively placed into the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 or the fourth incubation grid 5, the back of the PVDF membrane is attached to the bottom end of the inner wall of the box body 1, and then the antibody diluent of the corresponding type is dripped on the PVDF membrane according to the protein to be detected. The incubation of antibodies of different width bands is facilitated by the first incubation grid 2, the second incubation grid 3, the third incubation grid 4 and the fourth incubation grid 5, which are different in size. Through lid 6, be convenient for first hatch check 2, the second is hatched check 3, the third is hatched check 4 and the fourth is hatched 5 open-top covers of check to be convenient for guarantee that the antibody in the box body 1 can not volatilize in a large number. The design of the side wall of the box body is combined with the super-hydrophobic nano coating 7, so that the situation that the antibody splashes out of the surface of the PVDF membrane due to collision in the experimental process and cannot fall back to the membrane to cause experimental failure is conveniently prevented. Meanwhile, through the super-hydrophobic nano layer, the box body 1 is convenient to clean, and pollution to subsequent experiments can be avoided, so that water resources are saved to a certain extent, and the environment-friendly effect is achieved. Through first hatching check 2, second hatching check 3, the third is hatched check 4, the check 5 is hatched to the fourth, the box is hatched to the scour protection that check 6 and super hydrophobic nanometer layer constitute and is hit accuse temperature antibody, can prevent that the antibody diluent from splashing when box body 1 is collided among the experimentation and making the membrane of placing in 1 bottom of box body dry on hatching the box wall, thereby can guarantee the result of anti hatching, and can reduce time and the expenditure among the experimentation effectively.

The present embodiment is only for explaining the present invention, and it is not limited to the present invention, and those skilled in the art can make modifications to the present embodiment without inventive contribution as required after reading the present specification, but all of them are protected by patent laws within the scope of the claims of the present invention.

Claims

1. Anti-collision temperature-control antibody incubation box, which comprises a box body (1) and is characterized in that: the box body (1) is composed of a first incubation grid (2), a second incubation grid (3), a third incubation grid (4) and a fourth incubation grid (5), wherein the bottom surface and the top surface are positioned on the same plane and are different in size; the box body (1) is provided with a box cover (6) for covering the top opening of the box body (1); the inner wall of the box body (1) and the bottom surface of the box cover (6) are provided with super-hydrophobic nano coatings (7).

2. The anti-collision temperature-controlled antibody incubation box of claim 1, which is characterized in that: the first incubation grids (2), the second incubation grids (3), the third incubation grids (4) and the fourth incubation grids (5) are inverted truncated pyramid bodies; the end of the first incubation grid (2) with the top end open is connected with the end of the second incubation grid (3) with the top end open, the end of the second incubation grid (3) with the top end far away from the first incubation grid (2) is connected with the end of the third incubation grid (4) with the top end open, and the end of the third incubation grid (4) with the top end far away from the second incubation grid (3) is connected with the end of the fourth incubation grid (5) with the top end open.

3. The anti-collision temperature-controlled antibody incubation box of claim 1, which is characterized in that: the length of the top end of the box body (1) is 13.0cm, and the width of the top end of the box body is 9.4 cm; the side surface of the box body (1) is in an inverted trapezoid shape, and the height of the box body (1) is 1.7 cm; the widths of openings at the top ends of the first incubation grid (2), the second incubation grid (3), the third incubation grid (4) and the fourth incubation grid (5) are 4.0cm, 3.5cm, 3.0cm and 2.5cm respectively; the lengths of the bottom surfaces of the first incubation grid (2), the second incubation grid (3), the third incubation grid (4) and the fourth incubation grid (5) are 8.4 cm; the widths of the bottom surfaces of the first incubation grid (2), the second incubation grid (3), the third incubation grid (4) and the fourth incubation grid (5) are 3.0cm, 2.5cm, 2.0cm and 1.5cm respectively; the distance between the bottom surfaces of the first incubation grid (2), the second incubation grid (3), the third incubation grid (4) and the fourth incubation grid (5) is 1.0 cm.

4. The anti-collision temperature-controlled antibody incubation box of claim 1, which is characterized in that: the cross section of the box cover (6) is rectangular, the length of the box cover (6) is 10.1cm, and the width of the box cover (6) is 9.5 cm; the bottom surface of the box cover (6) is provided with a protruding interface (11) which is matched with the openings at the top ends of the first incubation grid (2), the second incubation grid (3), the third incubation grid (4) and the fourth incubation grid (5), and the protruding interface (11) is rectangular.

5. The anti-collision temperature-controlled antibody incubation box of claim 1, which is characterized in that: the side walls of the first incubation grids (2), the second incubation grids (3), the third incubation grids (4) and the fourth incubation grids (5) are respectively provided with a refrigerating sheet (8) and a film type temperature sensor (9) which are used for controlling the temperature in the box body, and the temperature control range of the refrigerating sheet (8) is 4-37 ℃; the box body (1) is provided with a singlechip (10) connected with a refrigerating sheet (8) and a film type temperature sensor (9); the side wall of the box body (1) is provided with a solar cell (12).