CN116136511A - Anti-falling fixed comet electrophoresis device and use method thereof - Google Patents

Abstract

The invention provides a fixed comet electrophoresis device with anti-falling function and a using method thereof. The invention comprises a quartz containing container groove, a bottom quartz plate and a top quartz cover plate, wherein the bottom quartz plate is used as a bearing container of agarose syrup, the top quartz cover plate is provided with holes arranged in an array, the top quartz cover plate is detachably connected with the bottom quartz plate, the bottom of the bottom quartz plate and the bottom surface of the quartz containing container groove are provided with adjustable preset distances, and the invention further comprises a lifting mechanism which is used for carrying out lifting operation on the bottom quartz plate. The comet electrophoresis structure assembly structure provided by the invention can be used for simultaneously placing a plurality of groups of cell suspensions in different modes, and the plurality of groups are simultaneously processed, so that the uniformity in the pretreatment process is ensured.

Description

Anti-falling fixed comet electrophoresis device and use method thereof

Technical Field

The invention relates to the technical field of molecular biology, in particular to a fixed comet electrophoresis device with anti-falling function and a using method thereof.

Background

Comet electrophoresis mainly comprises a neutral single-cell gel electrophoresis technology for measuring DNA double-strand breaks and an alkaline single-cell gel electrophoresis technology for measuring DNA single-strand breaks. The method is characterized in that single cells are suspended in agarose gel, after being subjected to cracking treatment, electrophoresis is carried out in an electric field for a short time, fluorescent dye is used for dyeing, DNA degradation fragments formed in apoptotic cells are subjected to electrophoresis in the electric field, the electrophoresis speed is high in the electric field, cell nuclei show a comet pattern, and normal nuclei without DNA fracture keep a sphere shape during electrophoresis, so that the method is a rapid and simple apoptosis detection method.

The patent 'comet electrophoresis gel spreading method' describes that biological information is reflected through extrusion length by whole migration of charged DNA in the use process, characterization is realized through the damage percentage of the DNA and the comet moment, and the possibility of degumming is reduced through a gel spreading method and a reagent adding flow.

The patent 'sealed temperature control electrophoresis apparatus' provides a sealed constant temperature process for electrophoresis in an electrophoresis apparatus, wherein the electrophoresis apparatus mainly relies on a sealed bin body, a constant temperature chamber and a sample inlet, and is heated by a sealed water bath, and the sealed constant temperature process for electrophoresis is realized by the cooperative operation of the temperature control chamber and the constant temperature chamber, but the patent lacks fixation and sealing of agarose syrup.

The existing comet electrophoresis device has the following defects: 1. at present, in the pretreatment process of comet electrophoresis, cell samples cannot achieve good consistency in the electrophoresis operation process due to improper fixation of agarose gel. 2. At present, the existing comet electrophoresis modes are all operation modes of a single glass slide, and the uniformity of multiple comet electrophoresis cannot be achieved. 3. In the process of taking the tablet, the colloid with cells is soaked in alkaline solution, and the small-block-shaped film is easy to fall off due to the greasiness of the alkaline solution. 4. Commercial comet electrophoresis often causes film to fall off because colloid in the use of electrophoresis cannot be fixed, and finally the film falling off cannot obtain an ideal comet electrophoresis pattern.

Disclosure of Invention

According to the technical problem, the anti-falling fixed comet electrophoresis device is provided. The invention can ensure that the cells obtain more reasonable results under the condition of measuring DNA damage. The invention adopts the following technical means:

the utility model provides a have fixed comet electrophoresis apparatus of anti-drop, includes quartz holding container groove and sets up in bottom quartz plate and top quartz cover plate wherein, bottom quartz plate is as the container that bears of agarose, be equipped with the hole that the array set up on the top quartz cover plate, top quartz cover plate detachably connects on the bottom quartz plate, the bottom of bottom quartz plate is equipped with adjustable preset distance with quartz holding container groove's bottom surface, still includes and carries and draws the mechanism, it is right to carry out to carry and draw the mechanism the bottom quartz plate carries out the operation.

Further, the adjustment of the preset distance between the bottom of the bottom quartz plate and the bottom surface of the quartz containing container groove is completed through the anti-skid wide-head screw, and screw holes are formed in four corners of the bottom quartz plate and the top quartz cover plate.

Further, the lifting mechanism comprises a lifting handle sheet metal part, the lifting handle sheet metal part comprises a handle part, a supporting part and a connecting part, the supporting part can be arranged in the quartz bearing container groove, the bottom quartz plate is placed on the supporting part, the handle part extends out of the quartz bearing container groove, the connecting part is connected with the supporting part and the handle part, and the supporting part enters into the quartz bearing container groove for a preset distance.

The invention also discloses a using method of the anti-falling fixed comet electrophoresis device, which comprises the following steps:

step 1, placing a bottom quartz plate and a top quartz cover plate in a quartz container tank, loading cracking liquid and neutralizing liquid in the quartz container tank, pouring agarose gel on the bottom quartz plate, covering the top quartz cover plate, and waiting for solidification for 10-20 minutes;

step 2, melting the low-melting-point agarose, preserving heat at 37 ℃ for later use, mixing cells with the melting-point agarose, uncovering a top quartz cover plate, and dripping the mixed colloid with the cells into a groove formed by the top quartz cover plate;

step 3, covering a cover glass with the same size as the groove, ensuring flatness, and setting at a certain temperature for solidification for 10-20 minutes;

step 4, the cover glass is torn off by the tweezers, then comet electrophoresis operation is carried out, and in the process, the handles are lifted, so that various pyrolysis liquids and neutralization liquids can be replaced conveniently;

and 5, placing the mixed sample with the glue and the cells on the quartz plate at the bottom in an electrophoresis tank, performing electrophoresis operation, cutting off the small square, and adding into fluorescent imaging.

The invention has the following advantages:

1. the comet electrophoresis structure assembly structure provided by the invention can be used for simultaneously placing a plurality of groups of cell suspensions in different modes, and the plurality of groups are simultaneously processed, so that the uniformity in the pretreatment process is ensured.

2. In the colloid extraction process, in order to avoid slipping of the alkaline solution and causing slipping of the films, the lifting handles are adopted to be more conducive to lifting up and down, and the small-unit colloid is in a block colloid, so that the extraction is convenient.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings may be obtained according to the drawings without inventive effort to a person skilled in the art.

FIG. 1 is a schematic diagram of the overall structure of the present invention.

Fig. 2 is an exploded front view of the component of the present invention.

Fig. 3 is an exploded perspective view of the components of the present invention.

Figure 4 is an exploded isometric view of the component of the present invention.

Fig. 5 is a diagram showing the practical application effect of the present invention.

In the figure: 001. a top quartz cover plate; 002. an anti-slip wide head screw; 003. a bottom quartz plate; 004. lifting the sheet metal part; 005. quartz holding container groove; 006. and lifting the handle connecting rod.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

As shown in fig. 1 to 4, the embodiment of the invention discloses an anti-falling fixed comet electrophoresis device, which comprises a quartz accommodating container groove 005, a bottom quartz plate 003 and a top quartz cover plate 001, wherein the bottom quartz plate 003 and the top quartz cover plate 001 are arranged in the container groove, the bottom quartz plate is used as an accommodating container of agarose syrup, holes arranged in an array are arranged on the top quartz cover plate, the top quartz cover plate is detachably connected with the bottom quartz plate, the bottom of the bottom quartz plate and the bottom surface of the quartz accommodating container groove are provided with adjustable preset distances, and the anti-falling fixed comet electrophoresis device further comprises a lifting mechanism, wherein the lifting mechanism is used for lifting the bottom quartz plate. The quartz containing container groove is used as a containing carrier of the pyrolysis liquid and the neutralization liquid.

The bottom of bottom quartz plate and the regulation of the bottom surface preset distance of quartz bearing container groove are accomplished through antiskid wide head screw 002, four bights of bottom quartz plate and top quartz cover plate all are equipped with the screw, the bottom quartz plate fills up certain height through cooperation top antiskid wide head screw, antiskid wide head screw also plays the effect of fixed colloid, prevents that the bulk sample from breaking away from the bottom quartz plate.

The lifting mechanism comprises a lifting handle sheet metal part 004, the lifting handle sheet metal part comprises a handle part, a supporting part and a connecting part, the supporting part can be arranged in a quartz bearing container groove, a quartz plate at the bottom is placed on the supporting part, the handle part extends out of the quartz bearing container groove, the connecting part is connected with the supporting part and the handle part, the supporting part enters into the quartz bearing container groove to be preset for a distance, the handle part and the supporting part are arranged in two sets and symmetrically, and a lifting handle connecting rod 006 is further arranged on the handle part in order to ensure the strength of the sheet metal part.

The invention also discloses a using method of the anti-falling fixed comet electrophoresis device, which comprises the following steps:

step 1, placing a bottom quartz plate and a top quartz cover plate in a quartz container tank, loading cracking liquid and neutralizing liquid in the quartz container tank, pouring agarose gel on the bottom quartz plate, covering the top quartz cover plate, and waiting for solidification for 10-20 minutes;

and step 2, after the digested cells are treated, the digested cells are ready for use. Melting low-melting-point agarose, preserving heat at 37 ℃ for later use, mixing cells with the melting-point agarose, uncovering a top quartz cover plate, and dripping the mixed colloid with the cells into a groove formed by the top quartz cover plate;

step 3, covering a cover glass with the same size as the groove, ensuring flatness, and setting at a certain temperature (4 ℃) for 10-20 minutes;

step 4, the cover glass is torn off by tweezers, and then comet electrophoresis normal operation is carried out, in the process, the handle is pulled up, and the replacement of various lysate and neutralizing solution is facilitated;

and 5, placing the mixed sample with the glue and the cells on the quartz plate at the bottom in an electrophoresis tank, performing electrophoresis operation, cutting off the small square lattice, and adding fluorescent imaging, wherein the specific experimental result is shown in figure 5.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.

Claims (4)

1. The utility model provides a have fixed comet electrophoresis apparatus of anti-drop, its characterized in that, include quartz bearing container groove and set up in bottom quartz plate and top quartz cover plate wherein, bottom quartz plate is as the bearing container of agarose, be equipped with the hole that the array set up on the top quartz cover plate, top quartz cover plate detachably connects on the bottom quartz plate, the bottom of bottom quartz plate is equipped with adjustable default distance with quartz bearing container groove's bottom surface, still includes and carries and draws the mechanism, it is right to carry and draw the mechanism carry out the operation to carry the bottom quartz plate.

2. The anti-falling fixed comet electrophoresis device of claim 1, wherein the adjustment of the preset distance between the bottom of the bottom quartz plate and the bottom surface of the quartz accommodating container groove is completed by anti-sliding wide-head screws, and four corners of the bottom quartz plate and the top quartz cover plate are respectively provided with screw holes.

3. The apparatus according to claim 1 or 2, wherein the pulling mechanism comprises a pulling handle sheet metal member comprising a handle portion, a supporting portion and a connecting portion, the supporting portion being capable of being disposed in a quartz receiving container groove, the bottom quartz plate being placed thereon, the handle portion extending out of the quartz receiving container groove, the connecting portion connecting the supporting portion and the handle portion, the supporting portion being entered into the quartz receiving container groove by a predetermined distance.

4. The application method of the fixed comet electrophoresis device with the anti-falling function is characterized by comprising the following steps of:

step 1, placing a bottom quartz plate and a top quartz cover plate in a quartz container tank, loading cracking liquid and neutralizing liquid in the quartz container tank, pouring agarose gel on the bottom quartz plate, covering the top quartz cover plate, and waiting for solidification for 10-20 minutes;

step 2, melting the low-melting-point agarose, preserving heat at 37 ℃ for later use, mixing cells with the melting-point agarose, uncovering a top quartz cover plate, and dripping the mixed colloid with the cells into a groove formed by the top quartz cover plate;

step 3, covering a cover glass with the same size as the groove, ensuring flatness, and setting at a certain temperature for solidification for 10-20 minutes;

step 4, the cover glass is torn off by the tweezers, then comet electrophoresis operation is carried out, and in the process, the handles are lifted, so that various pyrolysis liquids and neutralization liquids can be replaced conveniently;

and 5, placing the mixed sample with the glue and the cells on the quartz plate at the bottom in an electrophoresis tank, performing electrophoresis operation, cutting off the small square, and adding into fluorescent imaging.

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