CN112985958A - Negative pressure balance centrifugal bottle - Google Patents
Negative pressure balance centrifugal bottle Download PDFInfo
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- CN112985958A CN112985958A CN202110385315.2A CN202110385315A CN112985958A CN 112985958 A CN112985958 A CN 112985958A CN 202110385315 A CN202110385315 A CN 202110385315A CN 112985958 A CN112985958 A CN 112985958A
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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
- G01N1/34—Purifying; Cleaning
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Abstract
The invention provides a negative pressure balance centrifugal bottle, which comprises: a centrifuge bottle body and a fine opening with the inner diameter of 1-3 mm; the small opening is connected with the centrifuge bottle body through the conical body, and a section of thin tube is arranged between the small opening and the centrifuge bottle body. The negative pressure balance centrifugal bottle provided by the invention is arranged on a centrifugal machine, the centrifugal machine can effectively complete the whole processes of separation, cell concentration and flaking of cell specimen liquid through multiple times of high-low speed alternate operation, the flaking time is short, the time is greatly saved, the workload is reduced, the flaking efficiency is high, and the problem of cell loss caused by multiple times of cell specimen liquid transfer is effectively avoided.
Description
Technical Field
The invention relates to the technical field of cell sheet making equipment, in particular to a negative pressure balance centrifugal bottle.
Background
At present, the known exfoliated cell sheet preparation method usually needs to perform specific gravity separation on cell specimen liquid to centrifugally separate impurities, then further perform cell concentration centrifugation on the cell specimen liquid to collect cell sediments, then use a small amount of cell treatment liquid to enable the cell sediments to form cell suspension, and finally add the cell suspension with high concentration into a sheet preparation bin to prepare sheets.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a negative pressure balance centrifugal bottle, which comprises:
a centrifuge bottle body (1);
the centrifugal bottle comprises a fine opening (12), wherein the fine opening (12) is formed in one end of the centrifugal bottle body (1), and the inner diameter of the fine opening (12) is 1-3 mm.
Further, the fine opening (12) is connected with the centrifuge bottle body (1) through a conical body (14).
Further, a section of thin tube (13) is arranged between the small opening (12) and the centrifuge bottle body (1).
Further, still include joint portion (3), joint portion (3) set up tubule (13) or the surface of centrifugal flask body (1).
Further, the clamping part (3) is provided with a shoulder.
Further, the centrifugal bottle further comprises a bottle cap (11), wherein the bottle cap (11) is detachably arranged on the centrifugal bottle body (1), and the fine opening (12) is arranged on the bottle cap (11) or on the centrifugal bottle body (1).
Further, the end part of the small opening (12) is a detachable blind end (15), or the end part of the small opening (12) is provided with a detachable end cover (2).
Furthermore, the centrifuge bottle also comprises a partition (16) which is fixedly arranged in the centrifuge bottle body (1) and used for separating the sample liquid and the processing liquid, and a pore (161) is arranged on the partition.
Further, the partition (16) is in a conical funnel shape.
Furthermore, the partition (16) is flat-bottomed and is provided with a plurality of fine holes (161).
The negative pressure balance centrifugal bottle provided by the invention is clamped on the sheet making bin to form a centrifugal kit, the centrifugal kit is arranged on a centrifugal machine, the centrifugal machine can effectively complete the whole processes of specific gravity separation, cell concentration and sheet making of cell specimen liquid through multiple low-speed and high-speed circular rotations, the sheet making time is short, the time is greatly saved, the workload is reduced, the sheet making efficiency is high, and the problem of cell loss caused by multiple times of cell specimen liquid transfer is effectively solved.
Drawings
FIG. 1 is a schematic structural view of a negative pressure balancing centrifuge bottle provided by the present invention;
FIG. 2 is a schematic structural view of another negative pressure balancing centrifuge bottle provided by the present invention;
FIG. 3 is a schematic structural view of another negative pressure balancing centrifuge bottle provided by the present invention;
FIG. 4 is a schematic structural view of another negative pressure balancing centrifuge bottle provided in the present invention;
FIG. 5 is a schematic view of another negative pressure balancing centrifuge bottle of the present invention containing a specimen and a processing solution;
FIG. 6 is a schematic view of still another negative pressure balancing centrifuge bottle of the present invention containing a sample and a processing solution;
FIG. 7 is a schematic view of a conventional tablet making chamber;
FIG. 8 is a schematic structural diagram of a negative pressure balancing centrifuge bottle installed on a centrifuge according to the present invention;
FIG. 9 is a schematic structural diagram of the operation of the negative pressure balancing centrifuge bottle on the centrifuge;
wherein, 1, centrifuging the bottle body; 11. a bottle cap; 12. a fine opening; 13. a thin tube; 14. a conical body; 15. a blind end; 16. separating; 161. fine pores; 162. a second thin tube; 2. an end cap; 3. a clamping part; 31. a shoulder portion; 32. a guide portion; 4. a treatment liquid; 5. a cell sample solution; 6. a tablet making bin; 61. clamping a lug; 62. a seal ring; 7. a slide rack; 8. a glass slide; 9. a centrifugal shaft; 91. a centrifugal arm; 92. a hanging basket.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
In the description of the present invention, it is to be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplicity of description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention.
In the description of the present invention, it should be noted that unless otherwise explicitly stated or limited, the terms "mounted," "connected," and "disposed" are to be construed broadly and can, for example, be fixedly connected, disposed, detachably connected, disposed, or integrally connected and disposed. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
As shown in fig. 1, the present invention provides a negative pressure balancing centrifuge bottle comprising: centrifuge bottle body 1, bottle lid 11, tiny opening 12 with be provided with between the centrifuge bottle body 1 with tubule 13 and conical body 14 that tiny opening 12 links to each other, conical body 14's main aspects with centrifuge bottle body 1 links to each other, conical body 14's tip with tubule 13 links to each other, tubule 13's internal diameter can with tiny opening 12's internal diameter is the same, tiny opening 12's internal diameter is 1-3 mm.
The bottle cap 11 is detachably arranged on the centrifuge bottle body 1, so that the centrifuge bottle is sealed after the treatment fluid 4 and the cell specimen fluid 5 are added into the centrifuge bottle. The negative pressure balance centrifugal bottle fixing device further comprises a clamping portion 3, the clamping portion 3 is arranged on the outer surface of the thin tube 13, the centrifugal bottle body 1 or the conical body 14 and used for being matched with the sheet making bin 6 to facilitate the centrifugal bottle to be installed in the sheet making bin 6 to be positioned and supported, the clamping portion 3 is a positioning block with a convex shoulder, the shoulder 31 of the positioning block is matched with the end face of the centrifugal bin to facilitate the fixing of the negative pressure balance centrifugal bottle, and of course, the clamping portion 3 can be further provided with a guide portion 32 capable of extending into the centrifugal bin as shown in fig. 2 to facilitate the negative pressure balance centrifugal bottle to be installed in the centrifugal bin more stably.
The tip of tiny opening 12 is provided with detachable end cover 2, only need when the centrifugation film-making dismantle the end cover can.
In another embodiment of the negative pressure balancing centrifuge bottle, as shown in fig. 2, unlike the previous embodiment, the end of the small opening 12 is a blind end 15 of plastic or the like, which is easily cut off, and the blind end is cut off to expose the small opening during the centrifugal sheet making.
In another embodiment of the negative pressure balancing centrifuge bottle, as shown in fig. 3, unlike the previous embodiment, a thin tube 13 is provided on the bottle cap 11, and one end of a conical body 14 is connected to the thin tube 13 and the other end is connected to the centrifuge bottle body.
In another negative pressure balance centrifuge bottle, as shown in fig. 4, on the basis of the foregoing embodiment, the centrifuge bottle further comprises a partition 16 fixedly disposed in the centrifuge bottle body 1 for separating the sample liquid from the processing liquid, the partition 16 separates the centrifuge bottle body into an upper cavity and a lower cavity, and the partition is a conical funnel-shaped container with a second tubule 162 disposed at the bottom. The treatment solution 4 was added to the centrifuge bottle body 1, and then the cell sample solution 5 was added thereto by placing the partition 16, as shown in FIG. 5. The partition can play a role in separating the cell sample liquid from the treatment liquid, and reduce the pollution of the cell sample liquid to the treatment liquid.
In still another negative pressure balancing centrifuge bottle, as shown in fig. 6, the bottom of the partition 16 is provided with only a plurality of fine holes 161 without the second tubule, unlike the former centrifuge bottle. In FIG. 6, the treatment liquid 4 and the cell sample liquid 5 have been added.
Fig. 7 is a schematic structural diagram of a conventional tablet making chamber 6, which includes a hollow tubular structure with openings at two ends, a clamping lug 61 is disposed on an outer wall of one end of the tubular structure, and a sealing ring 62 protruding outwards is disposed on a port of one end of the tubular structure.
The method for performing centrifugal flaking by adopting the negative pressure balance centrifugal bottle provided by the invention comprises the following steps:
for convenience of description, another negative pressure balancing centrifuge bottle shown in fig. 2 is taken as an example for illustration,
(1) the bottle cap is opened, the treatment solution 4 and the cell specimen solution 5 are sequentially added into the centrifuge bottle, and then the bottle cap is tightly covered, so that a good sealing state is maintained between the bottle cap and the centrifuge bottle body, as shown in fig. 8. The density of the processing liquid is greater than that of the cell specimen liquid, so that the cell specimen liquid is suspended above the processing liquid, and a certain reserved space is reserved above the liquid level in the centrifuge bottle, namely the processing liquid and the cell specimen liquid do not occupy the accommodating space of the whole centrifuge bottle.
(2) An adhesive coated slide 8 is made and mounted in the slide holder 7.
(3) Insert the card ear in slide frame 7 in the film-making storehouse, the sealing washer in film-making storehouse 6 is hugged closely on slide 8, cut off the end cover on the centrifuge bottle and expose tiny opening 12, because the effect of surface tension, liquid in the centrifuge bottle can not follow tiny opening 12 and flow out in a large number, later will be equipped with treatment fluid 4, the centrifuge bottle of the mixed liquid of cell specimen liquid 5 installs and forms centrifugal external member on the other end in film-making storehouse, wherein tiny opening 12 on the centrifuge bottle towards the slide and with slide upper surface between the distance be 0.5-15mm, because joint portion is only some locating pieces, do not form sealing contact between centrifuge bottle and the film-making storehouse, film-making storehouse inside can keep unobstructed with external atmosphere.
(4) The centrifuge kit is placed in the basket 92 of a basket centrifuge. The basket centrifuge comprises a centrifugal shaft 9, a centrifugal arm 91 and a basket 92, wherein the centrifugal arm 91 is horizontally and fixedly arranged on the centrifugal shaft 9, the basket 92 is hinged on the centrifugal arm 91, as shown in figure 8,
(5) the centrifuge is started, the centrifuge shaft 9 rotates to drive the centrifuge arm 91 to rotate, the hanging basket 92 is gradually close to a horizontal state under the action of centrifugal force, and as shown in fig. 9, liquid in the centrifuge bottle flows into the tabletting bin under the action of centrifugal force.
The rotating speed of the centrifugal machine is gradually increased to the rotating speed N2And the liquid in the centrifugal bottle continuously flows into the tablet making bin under the action of centrifugal force along with the increase of the rotating speed for a certain time, and the liquid overflowsThe small opening 12 is formed, outside air can not enter the centrifugal bottle, the centrifugal bottle and the flaking bin form a communicating pipe with a single end opening, along with the continuous outflow of liquid, the space volume in the centrifugal bottle is continuously increased, the air pressure is continuously reduced, the negative pressure in the centrifugal bottle is gradually increased relative to the atmospheric pressure, the inner cavity of the flaking bin is communicated with the atmosphere, the liquid level in the flaking bin is always under the action of the atmospheric pressure, when the sum of the pressure generated by the air pressure in the centrifugal bottle and the centrifugal force of the liquid on the liquid level of the flaking bin is equal to the atmospheric pressure, namely, the pressure generated by the centrifugal force on the liquid level of the flaking bin is equal to the negative pressure in the centrifugal bottle, the liquid in the centrifugal bottle does not flow into the flaking bin any more, namely, the centrifugal force is balanced with the. Under the action of centrifugal force, cells, mucus and other particles continuously move into the treatment fluid and continuously move towards the flaking bin direction, and the cells move towards the flaking bin direction at a higher speed than the mucus due to the fact that the specific gravity of the cells is higher than that of other impurities such as the mucus, and other particles with lower specific gravity such as the mucus move towards the flaking bin direction at a lower speed.
(6) Rotational speed N2After the time is over, the rotating speed of the centrifugal machine is automatically increased to the rotating speed N1Maintaining for a certain time, breaking the balance between the negative pressure and the centrifugal force in the centrifugal bottle due to the increase of the centrifugal force, continuously flowing the liquid in the centrifugal bottle to the slide making bin, uniformly distributing the liquid to the periphery after entering the slide making bin, uniformly distributing the cells contained in the liquid to the periphery under the driving of the flowing liquid, and moving the cells in the slide making bin to the surface of the slide and adsorbing the cells by the slide under the action of the centrifugal force; along with the liquid constantly flows to the slide making storehouse, the negative pressure in the centrifuge bottle increases gradually and finally reaches the equilibrium with centrifugal force, and the liquid in the centrifuge bottle no longer flows into the slide making storehouse, and at this moment, although the total volume that the liquid flowed into the slide making storehouse no longer increases, particulate matters such as cell still continue to flow into the slide making storehouse through tiny opening, but after the cell reached the slide making storehouse, although the liquid in the slide making storehouse has stopped flowing, the cell that reaches in the slide making storehouse still has the chance to be adsorbed by the adhesive on the slide glass.
(7) Rotational speed N1After the time is over, the rotating speed of the centrifugal machine is automatically reduced to the rotating speed N2And maintained for a certain time becauseThe centrifugal force is reduced, the negative pressure in the centrifugal bottle is larger, and the liquid in the tablet making bin overflows the fine opening to form sealing, so that the liquid in the tablet making bin can flow back to the centrifugal bottle through the fine opening. When the liquid in the slide making bin flows back into the centrifugal bottle, the negative pressure in the centrifugal bottle is gradually reduced and finally is balanced with the centrifugal force, at the moment, the total volume of the liquid flowing back to the centrifugal bottle is not increased any more, and most of cells in the slide making bin are adsorbed by the slide glass, so that the concentration of the cells flowing back to the centrifugal bottle from the slide making bin is extremely low. At the same time, the cells in the centrifuge bottle are constantly moving toward the fine opening when the centrifuge bottle is continuously operated at a low rotation speed.
(8) Repeating the step (6) and the step (7) for a plurality of times, and finally enabling a sufficient number of cells to be uniformly adsorbed by the glass slide.
Wherein the rotating speed N1In the range of 200-2In the range of 100-900 rpm.
In another negative pressure balancing centrifuge bottle as shown in fig. 4, the separation wall is sealed with the inner wall of the centrifuge bottle body, and the separation wall divides the centrifuge bottle body into an upper chamber and a lower chamber, so that two times of negative pressure balancing are formed in the centrifuging process. Because the residual space after the upper chamber is filled with the cell sample liquid is very small, and the opening of the second tubule 162 is positioned in the processing liquid in the lower chamber, the air in the lower chamber can not enter the upper chamber, once a little liquid in the upper chamber enters the lower chamber, the upper chamber forms a larger negative pressure and is balanced with the centrifugal force quickly, although the cell sample liquid and the processing liquid have intersolubility, the structure can not pollute the processing liquid even if a large amount of body fluid samples are added.
In addition, as shown in FIG. 6, in another negative pressure balance centrifugal bottle, when the diameter of the pores 161, especially the pores with a diameter as small as 100-.
The invention well utilizes the surface tension of liquid in the tiny opening and the thin tube, combines the balance of centrifugal force and negative pressure, and uses high and low rotating speed N1、N2The alternate centrifugation method of (1), the cells are subjected to a plurality of elutriationsSelect and fully enter into the film-making storehouse and adhere to on the slide glass, be suitable for the sample liquid film-making of various concentration, various condition, adaptability is wide, and the film-making flow is simple, and the film-making is effectual.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
Claims (10)
1. A negative pressure balancing centrifuge bottle, comprising:
a centrifuge bottle body (1);
the centrifugal bottle comprises a fine opening (12), wherein the fine opening (12) is formed in one end of the centrifugal bottle body (1), and the inner diameter of the fine opening (12) is 1-3 mm.
2. Negative pressure balancing centrifuge bottle according to claim 1, characterized in, that the fine opening (12) is connected to the centrifuge bottle body (1) by means of a cone (14).
3. Negative pressure balancing centrifuge bottle according to claim 1, characterized in that a section of tubule (13) is arranged between the fine opening (12) and the centrifuge bottle body (1).
4. The negative pressure balance centrifuge bottle of claim 3, further comprising a clamping portion (3), wherein the clamping portion (3) is disposed on the outer surface of the tubule (13) or the centrifuge bottle body (1).
5. The negative pressure balancing centrifuge bottle of claim 4, wherein the snap-in portion (3) has a shoulder.
6. The negative pressure balancing centrifuge bottle of claim 1, further comprising a bottle cap (11), wherein the bottle cap (11) is detachably provided on the centrifuge bottle body (1), and the fine opening (12) is provided on the bottle cap (11) or on the centrifuge bottle body (1).
7. Negative pressure balancing centrifuge bottle according to claim 1, characterized in, that the end of the small opening (12) is a detachable blind end (15) or the end of the small opening (12) is provided with a detachable end cap (2).
8. The negative pressure balance centrifuge bottle according to claim 1, further comprising a partition (16) fixedly disposed in the centrifuge bottle body (1) for separating the sample liquid from the processing liquid, wherein the partition is provided with a fine hole (161).
9. Negative pressure balancing centrifuge bottle according to claim 8, characterized in, that the partition (16) is cone-shaped funnel-shaped.
10. The negative pressure balance centrifuge bottle of claim 8, wherein the partition (16) has a plurality of pores (161).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110385315.2A CN112985958A (en) | 2021-04-09 | 2021-04-09 | Negative pressure balance centrifugal bottle |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110385315.2A CN112985958A (en) | 2021-04-09 | 2021-04-09 | Negative pressure balance centrifugal bottle |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112985958A true CN112985958A (en) | 2021-06-18 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110385315.2A Pending CN112985958A (en) | 2021-04-09 | 2021-04-09 | Negative pressure balance centrifugal bottle |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112985958A (en) |
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2021
- 2021-04-09 CN CN202110385315.2A patent/CN112985958A/en active Pending
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