CN111678752A - Apoptosis detection kit capable of rapidly outputting result - Google Patents
Apoptosis detection kit capable of rapidly outputting result Download PDFInfo
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- CN111678752A CN111678752A CN202010570223.7A CN202010570223A CN111678752A CN 111678752 A CN111678752 A CN 111678752A CN 202010570223 A CN202010570223 A CN 202010570223A CN 111678752 A CN111678752 A CN 111678752A
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- 230000006907 apoptotic process Effects 0.000 title claims abstract description 19
- 238000001514 detection method Methods 0.000 title claims abstract description 16
- 238000001125 extrusion Methods 0.000 claims description 38
- 230000005540 biological transmission Effects 0.000 claims description 4
- 230000037431 insertion Effects 0.000 claims 8
- 238000003780 insertion Methods 0.000 claims 8
- 238000001556 precipitation Methods 0.000 abstract description 24
- 238000000034 method Methods 0.000 abstract description 6
- 239000007788 liquid Substances 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 27
- 238000004737 colorimetric analysis Methods 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 238000000149 argon plasma sintering Methods 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 230000001464 adherent effect Effects 0.000 description 2
- 230000001640 apoptogenic effect Effects 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000010428 chromatin condensation Effects 0.000 description 1
- 239000012916 chromogenic reagent Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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Classifications
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- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G01N15/01—
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Abstract
The invention discloses a cell apoptosis detection kit capable of rapidly outputting results, which is provided with a rotating shaft, a first gear and a second gear, wherein when a cell test sample is subjected to centrifugal precipitation work, the cell sample in a centrifuge tube is subjected to centrifugal precipitation, and the precipitation condition of the cell sample can be observed at any time in the centrifugal precipitation work process, so that the continuous precipitation work or the stop of the precipitation work can be better carried out according to whether the precipitation condition meets the requirement, precipitated cell liquid can be taken out in time, the cell test progress is accelerated, the position of the centrifuge tube can be well limited and fixed through a position fixing structure, the rapid proceeding of the centrifugal precipitation work is ensured, the precipitation deviation is prevented from generating, and the cell precipitation work efficiency is improved, thereby accelerating the working progress of cell precipitation and accelerating the generation of test results.
Description
Technical Field
The invention relates to the technical field of a device for detecting apoptosis of living cells, in particular to a kit for detecting apoptosis, which can rapidly output results.
Background
When the cell is apoptotic, due to cytoplasm and chromatin condensation and nuclear fragmentation, an apoptotic body is generated, so that the light scattering property of the cell is changed; early in apoptosis, the cells have significantly reduced ability to scatter forward light and increased or unchanged ability to scatter side light; in the late stage of apoptosis, signals of forward light scattering and side light scattering are reduced, so that the apoptosis condition can be observed by measuring the change of the light scattering of the cells by a flow cytometer; for use in a colorimetric apoptosis detection kit, the colorimetric method is based on a chromogenic reaction that produces a colored compound, and generally comprises two steps: firstly, selecting a proper chromogenic reagent to react with a component to be detected to form a colored compound, and then comparing or measuring the color depth of the colored compound; two colorimetric methods are commonly used in apoptosis detection kits: visual colorimetry and photoelectric colorimetry, both of which are based on lambert-beer law (a ═ bc); the common visual colorimetry is a standard series method, namely that standard solutions of objects to be measured with different quantities are firstly developed according to analysis steps in a group of colorimetric tubes which are completely the same to prepare a standard color gradation with gradually changing colors; the sample solution is developed under the same condition, and compared with the standard color gradation, the standard with the most similar color is visually found out, and the content of the component to be measured in the sample is calculated and determined according to the amount of the standard solution contained in the standard solution.
For adherent cells: carefully collecting the cell culture solution into a centrifugal tube for later use; digesting the cells by pancreatin, adding the previously collected cell culture solution when the cells can be blown down by a pipette or a gun head, blowing down all adherent cells, and blowing away the cells; again collected in the centrifuge tube. About 1000g, the cells are precipitated by centrifugation, some cells are centrifuged manually by workers in the precipitation process, so that the observation is convenient, but the working efficiency is too slow, the detection efficiency of a detection result is influenced, and moreover, the centrifuge tube is placed in a centrifuge for centrifugation, and in the process, the precipitation condition in the centrifuge tube cannot be observed at any time, so that the cells cannot be taken out in time for the next experimental detection when the centrifugation meets the requirement, the waste of the working time is further caused, and the test efficiency of the detection result is also influenced.
Disclosure of Invention
The invention aims to provide a kit for detecting apoptosis, which can rapidly output results, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a cell apoptosis detection kit capable of rapidly outputting results comprises a kit main body, a placing box and a position fixing structure;
a placing box is arranged at the side position of the kit main body, a first rotating groove is formed in the placing box, a rotating ball body is rotatably connected and arranged in the first rotating groove, a second rotating groove is rotatably connected and arranged at the other side of the rotating ball body, and the second rotating groove is arranged at one end of the rotating shaft;
the other end of the rotating shaft is provided with a first gear, the first gear is meshed with and connected with a second gear, the second gear is arranged at one end of the transmission shaft, the first gear is arranged on the side wall of one end of the supporting column, the other end of the supporting column is provided with a connecting column, the other end of the supporting column is provided with a limiting disc, and supporting rods are symmetrically arranged on the end face of the supporting column;
the support rod is movably connected with a movable rod, a first limiting block is arranged in the movable rod, a second limiting block is arranged in the support rod, grooves are formed in the movable rod at equal intervals, a first extrusion spring is arranged in each groove, a clamping block is connected to the other end of the first extrusion spring, the clamping blocks are movably connected and arranged in the grooves, first slopes are symmetrically arranged on the clamping blocks, a first extrusion block and a second extrusion block are inserted in the support rod, and a first reset spring is connected to the same end of the first extrusion block and the same end of the second extrusion block;
and the other end of the movable rod is provided with a position fixing structure.
Preferably, the position fixing structure comprises a placing circular tube, a fastening screw, a second extrusion spring, an extrusion piece, a first movable fixture block, a second reset spring, a sliding block, a clamping groove, a movable ring, a first insert block, a second insert block, a third reset spring, a fixed circular groove, a fixed block and a spring pin, wherein the movable rod is symmetrically arranged at one end of the placing circular tube, threaded holes are symmetrically arranged in the placing circular tube, the fastening screw is arranged in the threaded holes in a threaded connection manner, one end of the fastening screw is connected with the second extrusion spring, the other end of the second extrusion spring is connected with the extrusion piece, a first movable fixture block and a second movable fixture block are symmetrically inserted in the placing circular tube, the same end of the first movable fixture block and the second movable fixture block is symmetrically connected with the second reset spring, and a second slope is arranged at one end of the first movable fixture block, it is provided with the slider to place pipe avris position, and the equidistance is provided with the draw-in groove on the slider to the lateral wall cup joints and is provided with the loose collar, the avris of first inserted block is provided with fixed circular slot, the second inserted block other end is provided with the spout to first inserted block is provided with third reset spring with the same end symmetry of second inserted block, the avris of first inserted block is provided with fixed circular slot, the second inserted block other end, first inserted block avris position is provided with the fixed block, the one end that the intussuseption of fixed block was provided with spring catch, spring catch sets up the pull ring.
Preferably, the second gear and the first gear have the same tooth form angle and module, and the major diameter of the second gear is larger than that of the first gear.
Preferably, the number of the second movable clamping block and the fastening screw is the same, and the second movable clamping block and the fastening screw are arranged in four groups.
Preferably, the second insert block and the slot are arranged at corresponding positions, and the cross section of the second insert block is the same as that of the slot, that is, the cross sections of the second insert block and the slot are equal in length and width.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the rotating shaft, the first gear and the second gear are arranged, when the centrifugal precipitation work is carried out on the cell test sample, the cell sample in the centrifugal tube is subjected to centrifugal precipitation, and the precipitation condition of the cell sample can be observed at any time in the centrifugal precipitation work process, so that the continuous precipitation work or the stop of the precipitation work can be better carried out according to whether the precipitation condition meets the requirement, and then the precipitated cell liquid can be taken out in time, the centrifugal precipitation time is shortened, and the cell test progress is accelerated;
2. through the bracing piece that sets up, the swing joint of movable rod, can deposit cell centrifuging tube's size as required and carry out corresponding regulation, and then improve its application range to can carry out fine spacing and fixed to the position of centrifuging tube through the rigidity structure that sets up, guarantee going on fast of centrifugation sediment work, guarantee that it can not produce the sediment deviation, improve cell sediment work efficiency, and then accelerate cell sediment work progress, accelerate the test result and produce.
Drawings
FIG. 1 is a schematic top view of a structural connection according to the present invention;
FIG. 2 is a schematic bottom view of the structural connection of the present invention;
FIG. 3 is a schematic view of the internal connection structure of the placing box 2 according to the present invention;
FIG. 4 is an exploded view of the connecting structure of the support rod and the movable rod according to the present invention;
FIG. 5 is a bottom view of the placement tube in structural connection with the movable ring in accordance with the present invention;
FIG. 6 is a side view of the present invention showing the connection between the placement tube and the movable ring;
FIG. 7 is a top view of the placement tube and movable ring structure of the present invention;
fig. 8 is an exploded view of the connection structure of fig. 7 according to the present invention.
In the figure: the kit body 1, place the box 2, the ball that rotates 3, pivot 4, first gear 5, second gear 6, support column 7, spliced pole 8, spacing dish 9, bracing piece 10, movable rod 11, first stopper 12, second stopper 13, first extrusion spring 14, fixture block 15, first extrusion piece 16, second extrusion piece 17, first reset spring 18, position fixed structure 19, place pipe 1901, fastening screw 1902, second extrusion spring 1903, extrusion piece 1904, first activity fixture block 1905, second activity fixture block 1906, second reset spring 1907, slider 1908, draw-in groove 1909, activity ring 1910, first inserted block 1911, second inserted block 1912, third reset spring 1913, fixed circular slot 1914, fixed block 1915, spring pin 1916.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below. The embodiments of the present invention, and all other embodiments obtained by a person of ordinary skill in the art without any inventive work, belong to the scope of protection of the present invention.
Referring to fig. 1 to 8, the present invention provides a technical solution: a cell apoptosis detection kit capable of rapidly outputting results comprises a kit main body 1, a placing box 2 and a position fixing structure 19;
a placing box 2 is arranged at the side position of the kit main body 1, a first rotating groove is formed in the placing box 2, a rotating ball body 3 is rotatably connected and arranged in the first rotating groove, a second rotating groove is rotatably connected and arranged at the other side of the rotating ball body 3, and the second rotating groove is arranged at one end of a rotating shaft 4;
a first gear 5 is arranged at the other end of the rotating shaft 4, a second gear 6 is arranged at the first gear 5 in a meshed connection mode, the second gear 6 is arranged at one end of a transmission shaft, a micro motor is arranged at the other end of the transmission shaft in a connected mode, the first gear 5 is arranged on the side wall of one end of a supporting column 7, a connecting column 8 is arranged at the other end of the supporting column 7, a limiting disc 9 is arranged at the other end of the supporting column 7, and supporting rods 10 are symmetrically arranged on the end face;
a movable rod 11 is movably connected in the support rod 10, a first limit block 12 is arranged in the movable rod 11, a second limit block 13 is arranged in the support rod 10, grooves are formed in the movable rod 11 at equal intervals, a first extrusion spring 14 is arranged in each groove, a fixture block 15 is connected to the other end of the first extrusion spring 14, the fixture blocks 15 are movably connected in the grooves, first slopes are symmetrically arranged on the fixture blocks 15, a first extrusion block 16 and a second extrusion block 17 are inserted in the support rod 10, and a first return spring 18 is connected to the same end of the first extrusion block 16 as the second extrusion block 17;
the other end of the movable rod 11 is provided with a position fixing structure 19.
Further, the position fixing structure 19 includes a placing circular tube 1901, a fastening screw 1902, a second extrusion spring 1903, an extrusion sheet 1904, a first movable fixture block 1905, a second movable fixture block 1906, a second return spring 1907, a slider 1908, a clamping groove 1909, a movable ring 1910, a first insert block 1911, a second insert block 1912, a third return spring 1913, a fixed circular groove 1914, a fixed block 1915 and a spring pin 1916, wherein the movable rod 11 is symmetrically arranged at one end of the placing circular tube 1901, threaded holes are symmetrically arranged in the placing circular tube 1901, fastening screws 1902 are threadedly connected in the threaded holes, a second extrusion spring 1903 is connected at one end of the fastening screw 1902, an extrusion sheet 1904 is connected at the other end of the second extrusion spring 1903, a first movable fixture block 1905 and a second movable fixture block 1906 are symmetrically inserted in the placing circular tube 1901, a second return spring 1907 is symmetrically connected at the same end of the second movable fixture block 1906 of the first movable fixture block 1905, a second slope is arranged at one end of the first movable fixture block 1905, a slider 1908 is arranged at the side position of the round placing tube 1901, slots 1909 are equidistantly arranged on the slider 1908, a movable ring 1910 is sleeved on the outer side wall of the first movable fixture block 1905, a first inserting block 1911 and a second inserting block 1912 are inserted in the movable ring 1910, a third return spring 1913 is symmetrically arranged at the same end of the first inserting block 1911 and the second inserting block 1912, a fixed circular groove 1914 is arranged at the side of the first inserting block 1911, a sliding groove is arranged at the other end of the second inserting block 1912, a fixed block 1915 is arranged at the side position of the first inserting block 1911, and a spring pin 1916 and a pull ring are arranged at one end of the spring pin 1916 which are inserted in the fixed block 191.
Further, the second gear 6 and the first gear 5 have the same tooth profile angle and the same module, and the major diameter of the second gear 6 is larger than that of the first gear 5.
Further, the second movable latch 1906 and the fastening screw 1902 are provided in the same number of sets, and are provided in four sets.
Further, the second insert 1912 and the slot 1909 are disposed at corresponding positions, and the cross-sectional shape of the second insert 1912 is the same as the cross-sectional shape of the slot 1909, that is, the cross-sectional shapes are equal in length and width.
The working principle is as follows: when the invention is used for apoptosis test work, when centrifugal sedimentation work needs to be carried out on sample cells, a centrifugal tube is directly placed in a position fixing structure 19 on a kit main body 1 and a box 2, the centrifugal tube is inserted into the placing circular tube 1901, the bottom end of the centrifugal tube is contacted with a limiting disc 9, then a movable ring 1910 is pressed down to reach a proper position, namely, the position of the centrifugal tube is limited and fixed, a first inserting block 1911 is pressed, the first inserting block 1911 drives a second inserting block 1912 to be clamped in a clamping groove 1909 on a sliding block 1908, a fixed circular groove 1914 arranged on one side of the first inserting block 1911 reaches the position of a spring pin 1916, the spring pin 1916 is inserted and fixed in the fixed circular groove 1914, so that the position of the centrifugal tube is fixed, then a micro motor on the placing box 2 is started, a first gear 5, a second gear 6 and a rotating shaft 4 are driven to operate, so that centrifugal tube work can be carried out, realize the work of deposiing in the cell test, and in this in-process, the staff can the direct observation cell condition of deposiing, when deposiing and reaching the test work requirement, can take out it, it is convenient, and fast, and bracing piece 10 through the setting, the swing joint of movable rod 11, the size that can deposit the cell centrifuging tube as required carries out corresponding regulation, and then improve its application range, and can carry out fine spacing and fixed to the position of centrifuging tube through the fixed position structure that sets up, guarantee going on fast of centrifugal precipitation work, guarantee that it can not produce and deposit the deviation, improve cell precipitation work efficiency, and then accelerate cell precipitation progress, accelerate the test result and produce.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A cell apoptosis detection kit capable of rapidly outputting results is characterized in that: comprises a kit main body (1), a placing box (2) and a position fixing structure (19);
a placing box (2) is arranged at the side position of the kit main body (1), a first rotating groove is formed in the placing box (2), a rotating ball body (3) is rotatably connected and arranged in the first rotating groove, a second rotating groove is rotatably connected and arranged at the other side of the rotating ball body (3), and the second rotating groove is arranged at one end of a rotating shaft (4);
a first gear (5) is arranged at the other end of the rotating shaft (4), a second gear (6) is arranged on the first gear (5) in a meshed connection mode, the second gear (6) is arranged at one end of the transmission shaft, the first gear (5) is arranged on the side wall of one end of the supporting column (7), a connecting column (8) is arranged at the other end of the supporting column (7), a limiting disc (9) is arranged at the other end of the supporting column, and supporting rods (10) are symmetrically arranged on the end face of the supporting column (7);
a movable rod (11) is movably connected in the support rod (10), a first limit block (12) is arranged in the movable rod (11), a second limit block (13) is arranged in the support rod (10), grooves are formed in the movable rod (11) at equal intervals, a first extrusion spring (14) is arranged in each groove, a fixture block (15) is connected to the other end of each first extrusion spring (14), the fixture blocks (15) are movably connected in the grooves, first slopes are symmetrically arranged on the fixture blocks (15), a first extrusion block (16) and a second extrusion block (17) are inserted in the support rod (10), and a first return spring (18) is connected to the same end of the first extrusion block (16) and the second extrusion block (17);
the other end of the movable rod (11) is provided with a position fixing structure (19).
2. The rapid-outcome apoptosis detection kit according to claim 1, wherein: the position fixing structure (19) comprises a placing circular tube (1901), a fastening screw (1902), a second extrusion spring (1903), an extrusion sheet (1904), a first movable clamping block (1905), a second movable clamping block (1906), a second reset spring (1907), a sliding block (1908), a clamping groove (1909), a movable ring (1910), a first inserting block (1911), a second inserting block (1912), a third reset spring (1913), a fixed circular groove (1914), a fixed block (1915) and a spring pin (1916), wherein the movable rod (11) is symmetrically arranged at one end of the placing circular tube (1901), threaded holes are symmetrically arranged in the placing circular tube (1901), the fastening screw (1902) is arranged in the threaded holes in a threaded connection mode, the second extrusion spring (1903) is arranged at one end of the fastening screw (1902) in a connection mode, the extrusion sheet (1904) is arranged at the other end of the second extrusion spring (1903), and the first movable clamping block (1905) and the second movable clamping block are symmetrically arranged in an insertion mode in the placing circular tube (1901) (1906) The first movable fixture block (1905) is symmetrically connected with a second return spring (1907) at the same end of the second movable fixture block (1906), a second slope is arranged at one end of the first movable fixture block (1905), a sliding block (1908) is arranged at the edge side of the circular placement tube (1901), clamping grooves (1909) are equidistantly arranged on the sliding block (1908), a movable ring (1910) is sleeved on the outer side wall of the sliding block, a first insertion block (1911) and a second insertion block (1912) are arranged in the movable ring (1910) in an inserting mode, a third return spring (1913) is symmetrically arranged at the same end of the first insertion block (1911) and the second insertion block (1912) in an inserting mode, a fixed circular groove (1914) is arranged at the edge side of the first insertion block (1911), a sliding groove is arranged at the other end of the second insertion block (1912), a fixed block (1915) is arranged at the edge side of the first insertion block (1911), and a spring pin (1916) is arranged in an inserting mode, One end of the spring pin (1916) is provided with a pull ring.
3. The rapid-outcome apoptosis detection kit according to claim 1, wherein: the second gear (6) and the first gear (5) have the same tooth form angle and module, and the major diameter of the second gear (6) is larger than that of the first gear (5).
4. The rapid-outcome apoptosis detection kit according to claim 2, wherein: the number of the second movable fixture block (1906) and the number of the fastening screws (1902) are the same, and the second movable fixture block and the fastening screws are arranged in four groups.
5. The rapid-outcome apoptosis detection kit according to claim 2, wherein: the second insert block (1912) and the card slot (1909) are arranged at corresponding positions, and the cross section of the second insert block (1912) is the same as that of the slot of the card slot (1909), namely the cross sections are equal in length and width.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010570223.7A CN111678752B (en) | 2020-06-21 | 2020-06-21 | Apoptosis detection kit capable of rapidly outputting result |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010570223.7A CN111678752B (en) | 2020-06-21 | 2020-06-21 | Apoptosis detection kit capable of rapidly outputting result |
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| Publication Number | Publication Date |
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| CN111678752A true CN111678752A (en) | 2020-09-18 |
| CN111678752B CN111678752B (en) | 2021-02-26 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202010570223.7A Active CN111678752B (en) | 2020-06-21 | 2020-06-21 | Apoptosis detection kit capable of rapidly outputting result |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114994346A (en) * | 2022-08-04 | 2022-09-02 | 天津长和生物技术有限公司 | Cell detection device and detection method thereof |
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