A negative pressure balanced centrifuge bottle

Abstract

The present invention provides a negative pressure balanced centrifuge bottle, comprising: a centrifuge bottle body and a small opening with an inner diameter of 1 to 3 mm; the small opening is connected to the centrifuge bottle body via a conical body, and a thin tube is disposed between the small opening and the centrifuge bottle body. The negative pressure balanced centrifuge bottle provided by the present invention is installed in a centrifuge, and the centrifuge can effectively complete the entire process of separating cell specimens, concentrating cells, and preparing sections by repeatedly running the centrifuge at high and low speeds. This shortens the preparation time, greatly saves time, reduces workload, and achieves high preparation efficiency. It also effectively avoids the problem of cell loss caused by multiple transfers of cell specimens.

Description

Technical Field

[0001] The invention relates to the technical field of cell preparation equipment, in particular to a negative pressure balanced centrifuge bottle. Background Art

[0002] At present, the known exfoliated cell preparation method often requires first subjecting the cell specimen liquid to specific gravity separation to centrifuge to separate impurities, and then further subjecting the cell specimen liquid to cell concentration centrifugation to collect cell sediment, and then using a small amount of cell treatment fluid to make the cell sediment into a cell suspension, and finally adding the high-concentration cell suspension into the preparation chamber for preparation. Such a preparation method has many steps, a long preparation time, a large workload, and low efficiency, and the cell specimen liquid must be transferred multiple times. During the transfer process, cells are easily lost, thereby reducing the detection rate. Summary of the Invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention provides a negative pressure balanced centrifuge bottle, comprising:

[0004] Centrifuge bottle body (1);

[0005] A small opening (12) is provided at one end of the centrifuge bottle body (1), and the inner diameter of the small opening (12) is 1-3 mm.

[0006] Furthermore, the small opening (12) and the centrifuge bottle body (1) are connected via a cone (14).

[0007] Furthermore, a thin tube (13) is provided between the thin opening (12) and the centrifuge bottle body (1).

[0008] Furthermore, it also includes a clamping portion (3), and the clamping portion (3) is arranged on the outer surface of the capillary tube (13) or the centrifuge bottle body (1).

[0009] Furthermore, the clamping portion (3) has a shoulder.

[0010] Furthermore, the invention further comprises a bottle cap (11), wherein the bottle cap (11) is detachably arranged on the centrifuge bottle body (1), and the small opening (12) is arranged on the bottle cap (11) or on the centrifuge bottle body (1).

[0011] Furthermore, 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 cover (2).

[0012] Furthermore, it also includes a partition (16) fixedly arranged in the centrifuge bottle body (1) for separating the specimen liquid and the processing liquid, and the partition is provided with fine holes (161).

[0013] Furthermore, the partition (16) is in the shape of a conical funnel.

[0014] Furthermore, the partition (16) is flat-bottomed and has a plurality of pores (161) formed thereon.

[0015] The negative pressure balanced centrifuge bottle provided by the present invention is clamped on the preparation chamber to form a centrifuge kit. The centrifuge kit is installed on the centrifuge. The centrifuge can effectively complete the entire process of specific gravity separation, cell concentration and preparation of cell specimen liquid by multiple low-speed and high-speed cycle rotations. The preparation time is short, which greatly saves time, reduces workload, has high preparation efficiency, and effectively avoids the problem of cell loss caused by multiple transfers of cell specimen liquid. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a schematic structural diagram of a negative pressure balanced centrifuge bottle provided by the present invention;

[0017] Figure 2 It is a structural schematic diagram of another negative pressure balanced centrifuge bottle provided by the present invention;

[0018] Figure 3 This is a schematic structural diagram of another negative pressure balanced centrifuge bottle provided by the present invention;

[0019] Figure 4 This is a structural schematic diagram of another negative pressure balanced centrifuge bottle provided by the present invention;

[0020] Figure 5 This is a schematic diagram of another negative pressure balanced centrifuge bottle provided by the present invention containing a specimen and a processing liquid;

[0021] Figure 6 This is a schematic diagram of another negative pressure balanced centrifuge bottle provided by the present invention containing a specimen and a processing liquid;

[0022] Figure 7 This is a structural diagram of an existing film production warehouse;

[0023] Figure 8 This is a structural schematic diagram of a negative pressure balanced centrifuge bottle provided by the present invention installed on a centrifuge;

[0024] Figure 9 This is a structural schematic diagram of a negative pressure balanced centrifuge bottle provided by the present invention running on a centrifuge;

[0025] Among them, 1. Centrifuge bottle body; 11. Bottle cap; 12. Small opening; 13. Capillary tube; 14. Conical body; 15. Blind end; 16. Partition; 161. Pore; 162. Second capillary tube; 2. End cap; 3. Snap-fit ​​part; 31. Shoulder; 32. Guide part; 4. Processing liquid; 5. Cell specimen liquid; 6. Preparation chamber; 61. Ear; 62. Sealing ring; 7. Slide rack; 8. Slide; 9. Centrifuge shaft; 91. Centrifuge arm; 92. Hanging basket. DETAILED DESCRIPTION

[0026] The following describes embodiments of the present invention in detail. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended only to explain the present invention and are not to be construed as limiting the present invention.

[0027] In the description of the present invention, it should be understood that the terms "center", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0028] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "installed," "connected," and "disposed" should be understood in a broad sense. For example, they may refer to fixed connection or disposition, detachable connection or disposition, or integral connection or disposition. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0029] like Figure 1 As shown, the present invention provides a negative pressure balanced centrifuge bottle, comprising: a centrifuge bottle body 1, a bottle cap 11, and a small opening 12. A capillary tube 13 connected to the small opening 12 and a conical body 14 are provided between the small opening 12 and the centrifuge bottle body 1. The large end of the conical body 14 is connected to the centrifuge bottle body 1, and the small end of the conical body 14 is connected to the capillary tube 13. The inner diameter of the capillary tube 13 may be the same as the inner diameter of the small opening 12, and the inner diameter of the small opening 12 is 1-3 mm.

[0030] The bottle cap 11 is detachably mounted on the centrifuge bottle body 1, facilitating the sealing of the centrifuge bottle after adding the treatment liquid 4 and the cell specimen liquid 5 into the centrifuge bottle. The bottle cap 11 further comprises a clamping portion 3, which can be mounted on the outer surface of the capillary 13, the centrifuge bottle body 1 or the conical body 14, and is used to cooperate with the film preparation chamber 6 to facilitate the installation of the centrifuge bottle in the film preparation chamber 6 for positioning and support. The clamping portion 3 is a positioning block with a convex shoulder, and its shoulder 31 cooperates with the end face of the centrifuge chamber to facilitate the fixing of the negative pressure balanced centrifuge bottle of the present invention. Of course, the clamping portion 3 can also be as follows: Figure 2 As shown, a guide portion 32 is provided that can extend into the centrifugal chamber so that the negative pressure balanced centrifuge bottle of the present invention can be installed in the centrifugal chamber more stably.

[0031] The end of the small opening 12 is provided with a detachable end cap 2, and during centrifugal tableting, only the end cap needs to be removed.

[0032] In another embodiment of a negative pressure balanced centrifuge bottle, Figure 2 As shown, different from the above embodiment, the end of the small opening 12 is a blind end 15 made of plastic products that can be easily cut off. The blind end is cut off to expose the small opening during centrifugal slicing.

[0033] In another embodiment of a negative pressure balanced centrifuge bottle, Figure 3 As shown, different from the above embodiment, the capillary tube 13 is provided on the bottle cap 11, and one end of the conical body 14 is connected to the capillary tube 13, and the other end is connected to the centrifuge bottle body.

[0034] In another negative pressure balanced centrifuge bottle, Figure 4 As shown, based on the above embodiment, it further includes a partition 16 fixedly set in the centrifuge bottle body 1 for separating the specimen liquid and the processing liquid. The partition 16 divides the centrifuge bottle body into an upper chamber and a lower chamber. The partition is a conical funnel-shaped container with a thin tube 162 at the bottom. The processing liquid 4 is added to the centrifuge bottle body 1, and then the partition 16 is placed in. The cell specimen liquid 5 is added. Figure 5 The partition can separate the cell specimen fluid and the processing fluid, reducing the contamination of the cell specimen fluid to the processing fluid.

[0035] In another negative pressure balanced centrifuge bottle, such as Figure 6 As shown, the difference from the previous centrifuge bottle is that the bottom of the partition 16 is only provided with a plurality of fine holes 161 without a fine tube 2. Figure 6 In the process, treatment solution 4 and cell specimen solution 5 have been added.

[0036] Figure 7It is a structural diagram of an existing film making chamber 6, which includes a hollow tubular structure with two ends open, an ear 61 is provided on the outer wall of one end of the tubular structure, and a sealing ring 62 protruding outward is provided on the port of one end of the tubular structure.

[0037] The method for centrifugal tableting using the negative pressure balanced centrifuge bottle provided by the present invention is as follows:

[0038] For ease of description, Figure 2 Another negative pressure balanced centrifuge bottle is shown as an example to illustrate,

[0039] (1) Open the bottle cap and add the treatment solution 4 and the cell specimen solution 5 into the centrifuge bottle in sequence. Then, close the bottle cap tightly to ensure a good sealing state between the bottle cap and the centrifuge bottle body. Figure 8 As shown in the figure, the density of the treatment fluid is greater than that of the cell specimen fluid. Therefore, the cell specimen fluid is suspended above the treatment fluid, and there is a certain reserved space above the liquid surface in the centrifuge bottle, that is, the treatment fluid and the cell specimen fluid do not occupy the entire holding space of the centrifuge bottle.

[0040] (2) A glass slide 8 coated with an adhesive is prepared and installed in a glass slide rack 7 .

[0041] (3) Insert the ear of the preparation chamber into the slide rack 7, and the sealing ring of the preparation chamber 6 is tightly attached to the slide 8. Cut off the end cap of the centrifuge bottle to expose the small opening 12. Due to the effect of surface tension, the liquid in the centrifuge bottle will not flow out of the small opening 12 in large quantities. Then, install the centrifuge bottle containing the mixed solution of the treatment liquid 4 and the cell specimen liquid 5 on the other end of the preparation chamber to form a centrifuge kit, wherein the small opening 12 on the centrifuge bottle faces the slide and the distance between it and the upper surface of the slide is 0.5-15mm. Since the clamping part is just some positioning blocks, there is no sealed contact between the centrifuge bottle and the preparation chamber, and the inside of the preparation chamber and the outside atmosphere can be kept unobstructed.

[0042] (4) The centrifuge kit is placed in the hanging basket 92 of the hanging basket centrifuge. The hanging basket centrifuge includes a centrifugal shaft 9, a centrifugal arm 91, and a hanging basket 92. The centrifugal arm 91 is fixedly mounted horizontally on the centrifugal shaft 9, and the hanging basket 92 is hinged on the centrifugal arm 91. Figure 8 As shown,

[0043] (5) Start the centrifuge, the centrifugal shaft 9 rotates and drives the centrifugal arm 91 to rotate. Under the action of centrifugal force, the hanging basket 92 gradually approaches the horizontal state, as shown in FIG. Figure 9 As shown, the liquid in the centrifuge bottle flows into the film preparation chamber under the action of centrifugal force.

[0044] The speed of the centrifuge gradually increases to speed N2 and maintains it for a certain period of time. As the speed increases, the liquid in the centrifuge bottle continuously flows into the film-making chamber under the action of centrifugal force. The liquid overflows the small opening 12, and the outside air cannot enter the centrifuge bottle. The centrifuge bottle and the film-making chamber form a connecting pipe with a single-end opening. As the liquid continues to flow out, the space volume in the centrifuge bottle continues to increase, and the air pressure continues to decrease. Relative to the atmospheric pressure, the negative pressure in the centrifuge bottle gradually increases, and the inner cavity of the film-making chamber is connected to the atmosphere. The liquid level in the film-making chamber is always affected by the atmospheric pressure. When the sum of the air pressure in the centrifuge bottle and the pressure generated by the centrifugal force of the liquid on the liquid surface in the film-making chamber is equal to the atmospheric pressure, that is, when the pressure generated by the centrifugal force on the liquid surface in the film-making chamber is equal to the negative pressure in the centrifuge bottle, the liquid in the centrifuge bottle no longer flows into the film-making chamber, that is, the centrifugal force and the negative pressure are balanced. Under the action of centrifugal force, cells, mucus and other particles continuously move into the processing liquid and continue to move toward the film preparation chamber. Since the specific gravity of cells is greater than that of other impurities such as mucus, cells move toward the film preparation chamber at a faster speed than mucus, while other particles with smaller specific gravity such as mucus move toward the film preparation chamber at a slower speed.

[0045] (6) After the speed N2 time is over, the centrifuge speed automatically increases to speed N1 and maintains for a certain period of time. Due to the increase in centrifugal force, the balance between the negative pressure and centrifugal force in the centrifuge bottle is broken, and the liquid in the centrifuge bottle continues to flow to the preparation chamber. After the liquid enters the preparation chamber, it is evenly distributed to the surroundings. The cells contained in the liquid are also evenly distributed to the surroundings under the drive of the flowing liquid. Under the action of centrifugal force, the cells in the preparation chamber move to the surface of the slide and are adsorbed by the slide; as the liquid continues to flow to the preparation chamber, the negative pressure in the centrifuge bottle gradually increases and eventually reaches a balance with the centrifugal force, and the liquid in the centrifuge bottle no longer flows into the preparation chamber. At this time, although the total volume of liquid flowing into the preparation chamber no longer increases, cells and other particles continue to flow into the preparation chamber through the small opening. However, after the cells reach the preparation chamber, although the liquid in the preparation chamber has stopped flowing, the cells arriving in the preparation chamber still have the opportunity to be adsorbed by the adhesive on the slide.

[0046] (7) After the speed N1 time is over, the centrifuge speed automatically decreases to speed N2 and maintains it for a certain period of time. As the centrifugal force decreases, the negative pressure in the centrifuge bottle is large. Since the liquid level in the preparation chamber overflows the small opening to form a seal, the liquid in the preparation chamber can flow back to the centrifuge bottle through the small opening. When the liquid in the preparation chamber flows back to the centrifuge bottle, the negative pressure in the centrifuge bottle gradually decreases and eventually balances with the centrifugal force. At this time, the total volume of liquid flowing back to the centrifuge bottle no longer increases. Since most of the cells in the preparation chamber have been adsorbed by the slide, the concentration of cells flowing back from the preparation chamber to the centrifuge bottle is extremely low. But at the same time, when running continuously at a low speed, the cells in the centrifuge bottle always move continuously toward the small opening.

[0047] (8) Repeat steps (6) and (7) multiple times until a sufficient number of cells are evenly adsorbed on the glass slide.

[0048] The speed N1 ranges from 200 to 1000 rpm, and the speed N2 ranges from 100 to 900 rpm.

[0049] In such Figure 4 In another negative pressure balanced centrifuge bottle, the principle differs from the aforementioned method in that a partition is sealed against the inner wall of the centrifuge bottle body, dividing it into an upper chamber and a lower chamber. This creates two negative pressure balances during the centrifugation process. Since the upper chamber has very little remaining space after the cell specimen fluid is added, and the opening of the second capillary 162 is located in the treatment fluid in the lower chamber, air in the lower chamber cannot enter the upper chamber. Therefore, once a small amount of fluid enters the lower chamber, a large negative pressure is formed in the upper chamber and quickly balances with the centrifugal force. Although the cell specimen fluid and the treatment fluid are miscible, this structure prevents contamination of the treatment fluid even when a large amount of body fluid specimen is added.

[0050] In addition, if Figure 6 As shown, in another negative pressure balanced centrifuge bottle, the plurality of pores 161 on the partition, especially when the pore diameter is small and reaches 100-1000 μm, are suitable for specimens with more impurities, such as sputum specimens, and play the role of tearing mucus and filtering.

[0051] The present invention makes good use of the surface tension of liquid in small openings and thin tubes, combines the balance of centrifugal force and negative pressure, and uses an alternating centrifugation method of high and low speeds N1 and N2. Cells are fully selected through multiple selections and enter the preparation chamber and adhere to the slide. It is suitable for preparation of specimen liquids of various concentrations and various conditions, has wide adaptability, simple preparation process, and good preparation effect.

[0052] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the claims and their equivalents.

Claims

1. A negative pressure balanced centrifuge bottle, characterized in that, include: Centrifuge bottle body (1); A small opening (12), wherein the small opening (12) is provided at one end of the centrifuge bottle body (1), and the inner diameter of the small opening (12) is 1-3 mm; The small opening (12) is connected to the centrifuge bottle body (1) via a cone (14); A thin tube (13) is provided between the small opening (12) and the centrifuge bottle body (1); The large end of the conical body (14) is connected to the centrifuge bottle body (1), and the small end of the conical body (14) is connected to the cap (13); the bottle cap (11) is detachably arranged on the centrifuge bottle body (1); 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); The method for centrifugal tableting by the negative pressure balanced centrifuge bottle is as follows: (1) Open the bottle cap and add the treatment solution and cell specimen solution into the centrifuge bottle in sequence. Then, close the bottle cap tightly to ensure a good seal between the bottle cap and the centrifuge bottle body. (2) preparing a glass slide coated with an adhesive and mounting it in a glass slide holder; (3) Insert the ear of the preparation chamber into the slide rack, with the sealing ring of the preparation chamber tightly attached to the slide, cut off the end cap of the centrifuge bottle to reveal a small opening. Due to the effect of surface tension, the liquid in the centrifuge bottle will not flow out of the small opening in large quantities. Then, install the centrifuge bottle containing the mixed solution of the treatment solution and the cell specimen solution on the other end of the preparation chamber to form a centrifuge kit, wherein the small opening on the centrifuge bottle faces the slide and the distance between it and the upper surface of the slide is 0.5-15 mm; (4) Place the centrifuge kit in the basket of the hanging basket centrifuge and start the centrifuge.

2. The negative pressure balanced centrifuge bottle according to claim 1, characterized in that It also includes a clamping portion (3), which is arranged on the outer surface of the capillary tube (13) or the centrifuge bottle body (1).

3. The negative pressure balanced centrifuge bottle according to claim 2, characterized in that The clamping portion (3) has a convex shoulder.

4. The negative pressure balanced centrifuge bottle according to claim 1, characterized in that The small opening (12) is arranged on the bottle cap (11) or on the centrifuge bottle body (1).

5. The negative pressure balanced centrifuge bottle according to claim 1, characterized in that It also includes a partition (16) fixedly arranged in the centrifuge bottle body (1) for separating the specimen liquid and the processing liquid, and the partition is provided with fine holes (161).

6. The negative pressure balanced centrifuge bottle according to claim 5, characterized in that The partition (16) is in the shape of a cone funnel.

7. The negative pressure balanced centrifuge bottle according to claim 5, characterized in that The partition (16) is provided with a plurality of fine holes (161).

Images