TW202307195A - Sperm sorting apparatus and method sorting a plurality of motile sperms from a semen sample - Google Patents

Abstract

Provided is a sperm sorting apparatus suitable for sorting a plurality of motile sperms from a semen sample and including a base and a first sorter located on the base. The base includes an accommodating space defined by matching a bottom wall with a surrounding wall. The first sorter includes a first kit and a first permeable membrane, wherein the first kit is provided with a first sleeve placed into the accommodating space, and the first sleeve is provided with a first inner sleeve surface and a first outer sleeve surface. The first permeable membrane is fixed to the first sleeve, and the first permeable membrane matches with inner wall surfaces of the bottom wall and the surrounding wall to define a first sample accommodating chamber for accommodating the semen sample. Therefore, the motile sperms can naturally move upwards to penetrate through the first permeable membrane, and thus, the sperm sorting quality can be improved.

Description

Sperm sorting device and method

The invention relates to a sperm sorting device and method.

Referring to Fig. 1, the sperm sorting system 10 of U.S. US10,422,737B2 patent comprises a housing 12 that is formed with an inlet 11, a collection chamber 13 that is arranged on this housing 12 and is positioned at this inlet 11 side, an installation The microfluidic chip 14 on the bottom side of the housing 12, a flow path 15 extending from the inlet 11 to the collection chamber 13, and a filter 16 arranged in the collection chamber 13, the collection chamber 13 There is a bottom chamber 17 close to the microfluidic chip 14 , and a top chamber 18 located beside the bottom chamber 17 and away from the microfluidic chip 14 . The bottom chamber 17 is used to collect the semen sample injected from the inlet 11 into the flow channel 15, and the motile sperm in the semen sample move upward through the filter 16 and enter the top chamber 18, thereby achieving the goal of sperm sorting. Effect.

Although the sperm sorting system 10 can sort sperm, there are the following problems:

(1) The circulation path 15 and the bottom chamber 17 are originally filled with air, and the filter 16 has a very small pore size for the passage of motile sperm, and the air permeability is poor. In this way, when the semen sample is injected from the inlet 11 The push pressure generated during the process of entering the circulation passage 15 and entering the bottom chamber 17 will pressurize the volumetric airflow of the bottom chamber 17, and the push pressure will push the semen sample entering the bottom chamber 17 to The top chamber 18 pushes, so that some of the less motile sperm are pushed through the filter 16 into the top chamber 18, causing not all of the sperm taken from the top chamber 18 to be motile sperm , leading to poor sperm sorting quality.

(2) Because the distance between the inlet 11 and the bottom chamber 17 is relatively long and has two turns, the semen sample must be injected from the inlet 11 into the circulation channel 15 with a syringe to generate push pressure The semen sample is pushed into the bottom chamber 17, which is inconvenient to operate.

(3) After the sorting operation is completed, in order to analyze the sorted sperm, the whole sperm sorting system 10 must be taken to a microscope for analysis, which is inconvenient to operate.

Therefore, the first object of the present invention is to provide a sperm sorting device that can improve the quality of sperm sorting.

The second object of the present invention is to provide a sperm sorting method that can improve the quality of sperm sorting.

Therefore, the sperm sorting device of the present invention is suitable for sorting several motile sperm from a semen sample, and includes a base and a sorting unit. The base includes a bottom wall, and a surrounding wall extending from the periphery of the bottom wall toward the top and surrounding an axis, the bottom wall cooperates with the surrounding wall to define an accommodating space with an opening, and the surrounding wall has An inner wall surface and a top surface; the sorting unit includes a first sorter positioned at the base, the first sorter has a first sleeve and a first permeable membrane, and the first sleeve has a The axis is used to place the first sleeve in the accommodating space, the first sleeve has a first inner cylindrical surface, and a first outer cylindrical surface surrounding the first inner cylindrical surface; wherein, the first The permeable membrane cooperates with the bottom wall and the inner wall of the surrounding wall to define a first sample chamber for accommodating the semen sample.

The effect of the sperm sorting device of the present invention is that the gas originally located in the first sample container can be squeezed out of the first sample container, so that the motile sperm with strong motility in the semen sample The ability to pass through the first permeable membrane naturally and upwardly can improve the quality of sperm sorting.

The sperm sorting method of the present invention comprises the following steps: (A) Prepare the sperm sorting device as described above; (B) provide a semen sample; (C) drop the semen sample into the accommodating space from the opening of the base; (D) Put the first sleeve of the first sorter into the accommodating space and position it, the first permeable membrane is located above the semen sample, and the semen sample is accommodated in the first sample in the body container, and the air in the first sample container is squeezed out of the first sample container; (E) Drop a first sorting liquid on the first permeable membrane; (F) Stand still for 3 minutes to 60 minutes. During the standing process, several motile sperm with strong motility in the semen sample will move upward and pass through the first micropores of the first permeable membrane to enter the first sorting chamber is mixed with the first sorting fluid; and (G) Take out the first sorting liquid and the motile sperm from the first permeable membrane.

The effect of the sperm sorting method of the present invention is that: the gas originally located in the first sample container can be squeezed out of the first sample container, so that the motile sperm with strong motility in the semen sample The ability to pass through the first permeable membrane naturally and upwardly can improve the quality of sperm sorting.

Before the present invention is described in detail, it should be noted that the relative terms used in the following descriptions, such as "top" and "bottom", are based on the orientation shown in each figure and the normal use orientation, and similar elements are denoted by the same number. In addition, although the "barrel" and "ring" used in the following description are shown as a circle in the drawing, this should not limit that the "barrel" and "ring" must be circular.

Referring to FIGS. 2 to 5 , the sperm sorting device 21 according to the first embodiment of the present invention is suitable for sorting out several motile sperm 110 from a semen sample 100 , and includes a base 3 and a sorting unit 4 .

The base 3 includes a bottom wall 31, and a surrounding wall 32 extending from the periphery of the bottom wall 31 toward the top and surrounding an axis I. The bottom wall 31 cooperates with the surrounding wall 32 to define an opening 331. The accommodating space 33 , the surrounding wall 32 has an inner wall surface 321 and a top surface 322 , and an overflow groove 323 is formed on the top surface 322 . In this embodiment, the inner wall surface 321 is a circular inclined surface extending from the bottom wall 31 along the axis I toward the top surface 322 and gradually increasing in diameter, and the accommodating space 33 is a truncated conical space with a wide top and a narrow bottom.

It should be noted that the surrounding wall 32 also has an outer wall surface 324 surrounding the inner wall surface 321 , and the base 3 further includes an indicator scale 34 disposed on the outer wall surface 324 .

The sorting unit 4 includes a first sorter 40 having a first sleeve 41 and a first permeable membrane 44 .

The first sleeve 41 has a first sleeve 42 around the axis I for being inserted into the accommodating space 33, and a sleeve 42 extending from the top end of the first sleeve 42 toward a direction away from the axis I for positioning. The first protruding ring 43 on the top surface 322 of the surrounding wall 32 .

The first sleeve 42 has a first inner cylindrical surface 421, a first outer cylindrical surface 422 surrounding the first inner cylindrical surface 421, and a first inner cylindrical surface 421 connected to the first outer cylindrical surface 422. The bottom surface 423 of the first cylinder. In this embodiment, the first sleeve 42 extends from the bottom end along the axis I toward the top end and gradually increases in diameter, which is a frusto-conical sleeve with a wide top and a narrow bottom.

The first protruding ring 43 has a ring top surface 431, a ring bottom surface 432 and a ring peripheral surface 433 connecting the ring top surface 431 and the ring bottom surface 432. The ring bottom surface 432 forms a ring extending from the first outer cylinder surface 422 to The first escape groove 434 of the circumferential surface 433 .

The first permeable membrane 44 is fixed on the first cylinder bottom surface 423 of the first sleeve 42, and cooperates with the first inner cylinder surface 421 to define a first sorting chamber 45 with an open top. The permeable membrane 44 has a plurality of first micropores 441 for the motile sperm 110 to pass through, and the material of the first permeable membrane 44 can be polycarbonate (Polycarbonate), polystyrene (Polystyrene), polyterephthalene Polyethylene terephthalate, Polymethyl methacrylate), Polypropylene, Polytetrafluoroethylene, Nylon and Cellulose esters etc. materials or engineering plastics, and the area of the first permeable membrane 44 is less than 180cm², the diameter of each first micropore 441 is between 2 μm and 50 μm, and the first micropores 441 are relatively to the first permeable membrane The porosity of 44 is 5% to 90%, and the range used in this embodiment is that the pore diameter is between 8 μm and 20 μm, and the porosity is 5% to 20%.

It is worth noting that the first transparent film 44 can be fixed on the bottom surface 423 of the first sleeve 42 by means of ultrasonic welding, general heat welding, infrared welding or using UV adhesive, or on the bottom surface of the first sleeve 42 . The first inner cylindrical surface 421 of the first sleeve 42, or fixed on the reduced-diameter inner ring extending from the bottom end of the first sleeve 42 toward the axis I, is not limited to be fixed on the first cylindrical bottom surface 423 .

In addition, the first transparent membrane 44 also has a fixed portion 442 fixed to the first sleeve 42 around the axis I, and a transparent portion 443 extending from the fixed portion 442 toward the axis I unevenly. The penetrating portion 443 can be in a convex shape, a concave shape, or a wave shape, thereby increasing the penetrating area.

When the first sleeve 42 of the first sleeve 41 is put into the accommodating space 33, the first permeable membrane 44 cooperates with the bottom wall 31 and the inner wall surface 321 of the surrounding wall 32 to define a space for accommodating The first sample chamber 211 of the semen sample 100 is placed, and the inner wall surface 321 of the surrounding wall 32 cooperates with the first outer cylindrical surface 422 of the first sleeve 42 to define a channel communicating with the first sample chamber. 211 and the first through-space 212 of the first escape groove 434 .

The volume of the first sample chamber 211 is between 12ul and 400ml, the distance between the first permeable membrane 44 and the bottom wall 31 is between 50um and 8.5cm, and the first transparent gap 212 is less than 3mm, The area of the bottom surface of the first transparent gap 212 parallel to the bottom wall 31 is 0.25 mm² to 600 mm².

In addition, the first sorter 40 also has a liquid extractor socket 46 disposed on the first inner cylindrical surface 421 of the first set 41 and located in the first sorting chamber 45, the liquid extractor socket 46 There is an insertion hole 461 for inserting and positioning a liquid extractor (not shown, such as a syringe), and the diameter of the insertion hole is between 4um and 5mm.

The sperm sorting method corresponding to the sperm sorting device 21 of the present invention comprises the following steps:

Step 1: Prepare the sperm sorting device 21 .

Step 2: Provide the semen sample 100 of about 0.5㏄ to 50㏄.

Step 3: drop the semen sample 100 into the accommodating space 33 from the opening 331 of the base 3 .

Step 4: Put the first sleeve 42 of the first sorter 40 into the accommodating space 33 and position it, the first permeable membrane 44 is located above the semen sample 100, and the semen sample 100 is contained placed in the first sample chamber 211, and the air in the first sample chamber 211 is squeezed out of the first sample chamber 211, or squeezed into the first transparent space 212, The semen sample 100 occupies at least 80% of the volume of the first sample chamber 211 , so that the motile sperm 110 can pass through the first permeable membrane 44 smoothly.

Step 5: Drop a first sorting liquid 120 about 5㏄ to 50㏄ on the first permeable membrane 44, and the first sorting liquid 120 will partially penetrate into the first sample chamber 211 to form a continuous liquid On the other hand, the first sorting solution 120 is any diluent suitable for sperm treatment.

Step 6: Stand still for 3 minutes to 60 minutes. During the standing process, the motile sperm 110 in the semen sample 100 will move upward for a maximum distance of about 15mm, and then pass through the first pass. The first micropores 441 of the permeable membrane 44 enter the first sorting chamber 45 and mix with the first sorting liquid 120 .

Step 7: Take out the first sorting solution 120 and the motile sperm 110 from the first permeable membrane 44 .

The motile sperm 110 with strong motility can be obtained through the above steps, because the present invention places the semen sample 100 in the accommodating space 33, and the first sleeve of the first set 41 42 into the accommodating space 33, the gas originally located in the first sample container 211 can be squeezed out of the first sample container 211, or squeezed into the first transparent space 212 When the gas exceeds the volume of the first transparent space 212, it can be further discharged to the outside through the first escape groove 434, so that the air pressure of the first sample chamber 211 will not be affected, so that the semen sample The motile sperm 110 with strong motility in the body 100 can freely pass upwards through the first permeable membrane 44 and enter the first sorting chamber 45. Therefore, the present invention obtains from the first permeable membrane 44 The motile sperm 110 are sperm with strong motility, which can achieve the effect of improving the quality of sperm sorting.

It is worth noting that:

(1) During the process of placing the semen sample 100 in the accommodating space 33, the operator can confirm the amount of the semen sample 100 by the indicator scale 34. If the operator accidentally drops the semen sample 100 If there are too many semen samples 100, the volume of the semen samples 100 exceeds the volume of the first sample chamber 211, and the semen samples 100 can enter the overflow tank 323 of the base 3 through the first transparent gap 212 In this way, the surrounding environment can be avoided from being polluted.

(2) The first inner cylinder surface 421 of the first sleeve 42 can be coated with hyaluronic acid (Hyaluronic acid) coating, progesterone (Progesterone) coating, or an infrared light bulb structure, a power coil structure, or around the The inner wall surface 321 of the wall 32 adds a bionic structure, thereby improving the sorting efficiency.

(3) When the total free distance of the motile sperm 110 is less than 15 mm, when the volume ratio of the first sample chamber 211 to the first sorting chamber 45 is less than or equal to 1:1, it can be obtained The preferred quantity recovery rate of the motile sperm 110 .

(4) The liquid extractor (not shown) is inserted and fixed by the liquid extractor socket 46 , so that the operator can stably operate the liquid extractor to improve efficiency.

After the above step six, the operator can take out the first sorter 40 from the base 3, and directly place the first sorter 40 on the microscope stage (not shown) to target the motile sperm. 110 for analysis.

Referring to FIG. 6 , the base 3 further includes an annular groove 35 disposed on the inner wall surface 321 of the surrounding wall 32 , and the first separator 40 also has a first inner cylindrical surface 421 disposed on the first sleeve 42 The first annular groove 47, the annular groove 35 and the first annular groove 47 have the function of preventing severe capillarity and blocking the expansion of capillarity, and the cross-sectional shape and setting of the annular groove 35 and the first annular groove 47 Height A is unlimited.

7 to 10, the sperm sorting device 22 of the second embodiment of the present invention includes the base 3, a sorting unit 4' and a top cover 60.

The sorting unit 4' includes two first sorters 40, wherein one of the first sorters 40 is placed on the base 3, both of which form the same structure as the sperm sorting device 21 (see Figure 4), The first sleeve 42 of another first sorter 40 is inserted into the first sleeve 42 of one of the first sorters 40, and the first protruding ring 43 of the other first sorter 40 is located in the first sleeve 42 of the first sorter 40. On the first protruding ring 43 of one of the first sorters 40, the two first sorters 40 are stacked along the axis I.

The top cover 60 includes a top wall 61, and a top surrounding wall extending from the periphery of the top wall 61 toward the bottom end and surrounding the axis I for being positioned on the first flange 43 of the first separator 40 62, the top wall 61 cooperates with the top surrounding wall 62 to define a top accommodating space 63 with an open bottom.

Regarding the use of the sperm sorting device 22, the operator can use the combined structure of the base 3 and a first sorter 40 to perform one layer of sperm sorting according to the needs of the user, or use the base 3 and two first sorting devices. The combined structure of the device 40 performs two-layer sperm sorting operations, so that sperm with stronger motility can be obtained on the first permeable membrane 44 of the other first sorter 40 . The method for the two-layer sperm sorting operation will be described in detail in the third embodiment below.

Regarding the top cover 60, the top surrounding wall 62 can be placed on the first protruding ring 43 of the first sorter 40 to close the first sorting chamber 45, or the top cover 60 can be topped and bottomed. Inverted, the top accommodating space 63 is opened toward the top, and the first sleeve 42 of the first sorter 40 is placed in the top accommodating space 63, so that the top cover 60 can also serve as a platform .

Referring to FIG. 11 to FIG. 13 , the sperm sorting device 23 according to the third embodiment of the present invention includes the base 3 , a sorting unit 4 ″ and the top cover 60 .

The sorting unit 4 ″ includes a first sorter 40 and a second sorter 50 .

The second separator 50 has a second set 51 and a second permeable membrane 55 .

The second sleeve 51 has a second sleeve 52 around the axis I for being inserted into the first sleeve 42, and a second sleeve 52 extending from the top end of the second sleeve 52 toward a direction away from the axis I for positioning. The second protruding ring 53 on the first protruding ring 43 , and an inner ring 54 extending from the bottom end of the second sleeve 52 toward the direction close to the axis I and tapered toward the top end.

The second sleeve 52 has a second inner cylindrical surface 521, a second outer cylindrical surface 522 surrounding the second inner cylindrical surface 521, and a second inner cylindrical surface 521 connected to the second outer cylindrical surface 522. The bottom surface 523 of the second cylinder. In this embodiment, the second sleeve 52 extends from the bottom end along the axis I toward the top end and gradually increases in diameter, which is a frusto-conical sleeve with a wide top and a narrow bottom.

The second convex ring 53 has a ring top surface 531, a ring bottom surface 532 and a ring peripheral surface 533 connecting the ring top surface 531 and the ring bottom surface 532. The ring bottom surface 532 forms a ring extending from the second outer cylinder surface 522 to The second air escape groove 534 of the circumferential surface 533 .

The inner ring 54 is a frusto-conical ring with a narrow top and a wide bottom.

The second permeable membrane 55 is fixed on the second cylinder bottom surface 523 of the second sleeve 52, and cooperates with the inner ring 54 and the second inner cylinder surface 521 to define a second sorting chamber with an open top 56, the second permeable membrane 55 has a plurality of second micropores 551 for several motile sperm 111 with stronger motility, the material of the second permeable membrane 55 is the same as the material of the first permeable membrane 44 Can be the same or different, the pore diameter of each second micropore 551 is between 2 μm and 50 μm, and the porosity of the second micropores 551 relative to the second permeable membrane 55 is 5% to 90%. In this embodiment The range used in the example is that the pore size is between 8 μm and 12 μm, and the porosity is 5% to 20%. It can be understood that the area size, material, pore size and porosity of the second permeable membrane 55 are not necessarily the same as that of the second permeable membrane 55. The first permeable membrane 44 is the same.

The second sorting chamber 56 is formed by the cooperation of the second permeable membrane 55 with the inner ring 54 of a truncated cone-shaped ring with a narrow top and a wide bottom, which can make the second sorting chamber 56 The volume is smaller than the volume of the first sorting chamber 45, thereby reducing the amount of sorting liquid used to reduce the cost of use.

Regarding the top cover 60, the top surrounding wall 62 can be placed on the second convex ring 53 of the second sorter 50 to close the second sorting chamber 56, or the top cover 60 can be topped and bottomed. Invert, make the top accommodating space 63 open toward the top, and put the first sleeve 42 of the first sorter 40 into the top accommodating space 63, or put the second sleeve 42 of the second sorter 50 into the top accommodating space 63. The sleeve 52 is put into the top accommodating space 63 , so that the top cover 60 also has the function of carrying platform.

When the first sleeve 42 of the first sorter 40 is put into the accommodating space 33, the first permeable membrane 44 cooperates with the bottom wall 31 and the inner wall surface 321 of the surrounding wall 32 to define a The first sample container chamber 211 for accommodating the semen sample 100, the inner wall surface 321 of the surrounding wall 32 cooperates with the first outer cylindrical surface 422 of the first sleeve 42 to define the first sample container. The first transparent gap 212 between the chamber 211 and the first escape groove 434, the first transparent gap 212 is 0.05 mm to 2 mm, when the second sleeve 52 of the second separator 50 is placed The first sorting chamber 45 of the first sorter 40, the first inner cylindrical surface 421 of the first sleeve 42 cooperates with the second outer cylindrical surface 522 of the second sleeve 52 to define a communication channel. The second transparent gap 232 between the first sorting chamber 45 and the second escape groove 534 is 0.05 mm to 2 mm. The sperm sorting method corresponding to the sperm sorting device 23 of the present invention comprises the following steps:

Step 1: Prepare the sperm sorting device 23 .

Step 2: Provide the semen sample 100 of about 0.5㏄ to 50㏄.

Step 3: drop the semen sample 100 into the accommodating space 33 from the opening 331 of the base 30 .

Step 4: Put the first sleeve 42 of the first sorter 40 into the accommodating space 33 and position it, the first permeable membrane 44 is located above the semen sample 100, and the semen sample 100 is contained placed in the first sample chamber 211, and the air in the first sample chamber 211 is squeezed out of the first sample chamber 211, or squeezed into the first transparent space 232, The semen sample 100 occupies at least 80% of the volume of the first sample chamber 211 , so that the motile sperm 110 can pass through the first permeable membrane 44 smoothly.

Step 5: Place a first sorting liquid 120 about 5㏄ to 100㏄ on the first permeable membrane 44, and the first sorting liquid 120 will partially permeate into the first sample chamber 211 to form a continuous liquid On the other hand, the first sorting solution 120 is any diluent suitable for sperm treatment.

Step 6: Put the second sleeve 52 of the second sorter 50 into the first sorting chamber 45, and place the second protruding ring 53 on the first protruding ring 43, the second channel The permeable membrane 55 is located above the first permeable membrane 44 .

Step 7: Drop a second sorting liquid 121 on the second permeable membrane 55 about 10 ㏄. The second sorting liquid 121 is any diluent suitable for sperm treatment.

Step 8: Stand still for 3 minutes to 60 minutes. During the standing process, the motile sperm 110 in the semen sample 100 will move upward for a maximum distance of 15mm and pass through the first penetrating The first micropores 441 of the membrane 44 enter the first sorting chamber 45 and mix with the first sorting liquid 120, and the motile sperm 111 with stronger motility on the first permeable membrane 44 will Freely pass through the second micropores 551 of the second permeable membrane 55 upwards, enter the second sorting chamber 56 and mix with the second sorting liquid 121 .

Step 9: Take out the second sorting solution 121 and the motile sperm 111 with stronger motility from the second permeable membrane 55 .

Through the above steps, the motile sperm 111 with stronger motility can be obtained, so that the present invention has the effect of improving the quality of sperm sorting.

It is worth noting that the sperm sorting device 23 can also be put the second sleeve 52 of the second sorter 50 into the accommodating space 33 of the base 3 first, and then the first sorter 40 The structure in which the first sleeve 42 is inserted into the second sleeve 52 of the second sorter 50 also has the effect of obtaining the motile sperm 111 with stronger motility.

In addition, the first sorting liquid 120 and the second sorting liquid 121 can be different sorting liquids.

Referring to FIG. 14 to FIG. 16 , the sperm sorting device 24 according to the fourth embodiment of the present invention includes the base 3 and a sorting unit 7 .

The sorting unit 7 includes the second sorter 50 .

When the second sleeve 52 of the second sorter 50 is put into the accommodating space 33, the second transparent membrane 55 cooperates with the bottom wall 31 and the inner wall surface 321 of the surrounding wall 32 to define a In the second sample container chamber 241 for accommodating the semen sample 100, the inner wall surface 321 of the surrounding wall 32 cooperates with the outer cylindrical surface 522 of the second sleeve 52 to define a channel communicating with the second sample container chamber. 241 and the third transparent gap 242 of the second escape groove 534, the third transparent gap 242 is 0.05 mm to 2 mm.

The sperm sorting device 24 has the same effect as the sperm sorting device 21 (see Figure 4), the difference is that the volume of the second sorting chamber 56 is smaller than the volume of the first sorting chamber 45, which can reduce The amount of sorting fluid used to reduce the cost of use.

17 to 19, the sperm sorting device 25 according to the fifth embodiment of the present invention includes the base 3, a sorting unit 7' and the top cover 60.

The sorting unit 7' comprises two second sorters 50 stacked along the axis I.

The top cover 60 can be placed on the second sorter 50, or it can be turned upside down, and the second sleeve 52 of the second sorter 50 is put into the top accommodating space 63, so that the top The cover 60 also functions as a stage.

The sperm sorting device 25 has the same function as the sperm sorting device 22 (see FIG. 9 ) and the sperm sorting device 23 (see FIG. 13 ).

Referring to FIGS. 20 to 22 , the sperm sorting device 26 according to the sixth embodiment of the present invention includes the base 3 , the sorting unit 4 and a height adjustment unit 8 .

The height adjustment unit 8 is arranged between the top surface 322 of the surrounding wall 32 of the base 3 and the ring bottom surface 432 of the first protruding ring 43 of the first sleeve 41, and includes Two first step portions 81, two second step portions 82 respectively connecting the two first step portions 81 and having a height along the axis I lower than the two first step portions 81 and diametrically opposite to each other, connecting the two second step portions The first step portion 81 and the two second step portions 82 and the third step portion 83 whose height along the axis I is lower than the two second step portions 82 and diametrically opposite to each other correspond to the first step portion 81, the The second step portions 82, the third step portions 83 are arranged on the ring bottom surface 432 and are two first seating portions 84 diametrically opposite to each other, and two first seating portions 84 respectively connected to the two first seating portions 84 and are diametrically opposite to each other. Two seat portions 85, and a third seat portion 86 connecting the two first seat portions 84 and the two second seat portions 85 and diametrically opposite to each other. The first steps 81, the second steps 82, and the third steps 83 are distributed at an angle of 60 degrees, and the first seats 84, the second seats 85, the third seats The seating portions 86 are distributed at an angle of 60 degrees. Both the top surface 322 and the ring bottom surface 432 are stepped surfaces.

When the two first seats 84 are seated on the two third steps 83, the two second seats 85 are seated on the two second steps 82, and the two third seats 86 are seated on the On the second first step portion 81, the first sleeve 41 is in a state of 0 degree rotation relative to the base 3. At this time, the sperm sorting device 26 has a first height along the axis I, and the first transparent gap 212 Having a first volume, the first sample chamber 211 has a first volume.

Referring to Fig. 23 and Fig. 24, when the first sleeve 41 is rotated 60 degrees relative to the base 3, the two first seat parts 84 are seated on the two second step parts 82, and the two second seat parts 85 are located on On the two first steps 81, the two third seats 86 are located on the two third steps 83, the sperm sorting device 26 has a second height along the axis I, and the first transparent gap 212 becomes large and has a second volume, the first sample chamber 211 has a second volume, and the second height is greater than the first height, the second volume is greater than the first volume, and the second volume is greater than The first capacity.

Referring to Fig. 25 and Fig. 26, when the first sleeve 41 is rotated 120 degrees relative to the base 3, the two first seat parts 84 are seated on the two first step parts 81, and the two second seat parts 85 are located on the On the second third step portion 83, the two third seating portions 86 are located on the second second step portion 82, the sperm sorting device 26 has a third height along the axis I, and the first transparent gap 212 becomes larger And has a third volume, the first sample chamber 211 has a third volume, and the third height is greater than the second height, the third volume is greater than the second volume, the third volume is greater than the Second capacity.

It should be noted that the height adjustment unit 8 does not have two first steps 81, two second steps 82, two third steps 83, two first seats 84, and two second steps. The number of the seat portion 85 and the two third seat portions 86 is limited, and other numbers of step portions and seat portions can also be provided. In addition, the arrangement of the above-mentioned step portion and the seat portion is not limited to 60 degrees and radially opposite each other. .

Referring to FIG. 27 and FIG. 28 , the sperm sorting device 27 according to the seventh embodiment of the present invention includes the base 3 , the sorting unit 4 and a height adjustment unit 9 .

The height adjustment unit 9 is arranged between the inner wall surface 321 of the surrounding wall 32 of the base 3 and the first outer cylindrical surface 422 of the first sleeve 42 of the first sleeve 41, and includes a The first inner thread segment 91 on the top surface 323 and the outer thread segment 92 on the first outer cylindrical surface 422 and close to the first protruding ring 43 .

By screwing the external threaded section 92 and the first internal threaded section 91 together, the first sorter 40 can be moved closer to or away from the base 3 , and the gap size of the first through space 212 can be adjusted accordingly. And the volume size of the first sample chamber 211 .

Referring to Fig. 29 and Fig. 30, the sperm sorting device 28 of the eighth embodiment of the present invention includes the base 3, the sorting unit 4' and a height adjusting unit 9'.

The height adjustment unit 9 ′ includes the first internal thread section 91 located on the inner wall surface 321 and close to the top surface 323 , the external thread section 92 located on the first outer cylinder surface 422 close to the first collar 43 , And a second internal thread section 93 located on the first inner cylindrical surface 421 of the first sleeve 42 of the first sleeve 41 and close to the first protruding ring 43 .

Through the threaded combination of the external threaded section 92 and the first internal threaded section 91, one of the first separators 40 can be moved closer to or farther away from the base 3, and then the first transparent gap 212 can be adjusted. The size of the gap and the volume of the first sample chamber 211 are screwed together by the external threaded section 92 of the other first sorter 40 and the second internal threaded section 93 of one of the first sorters 40. lock combination, the other first sorter 40 can be made close to or away from one of the first sorters 40, and then the volume of the first sorting chamber 45 of one of the first sorters 40 can be adjusted size.

Referring to FIG. 31 , the sperm sorting device 29 according to the ninth embodiment of the present invention includes the base 3 , the sorting unit 4 and a spacer 95 .

The spacer 95 is installed between the top surface 322 of the base 3 and the ring bottom surface 432 of the first convex ring 43 of the first separator 40, and the first separator 40 can be kept away from the first separator 40 by the gasket 95. The base 3 can further increase the clearance of the first transparent space 212 and increase the volume of the first sample chamber 211 .

Referring to Fig. 32 and Fig. 4, the sperm sorting device 21' of the tenth embodiment of the present invention and the sperm sorting device 21 also include the base 3 and the sorting unit 4, the difference between the two is that the sperm sorting device 21 The first protruding ring 43 is positioned on the top surface 322 of the surrounding wall 32 to position the first sorter 40, and the first sleeve 41 of the sperm sorting device 21' omits the first protruding ring 43. Since the accommodating space 33 is a truncated conical space with a wide top and a narrow bottom, and the first sleeve 42 is a truncated conical sleeve with a wide top and a narrow bottom, the first sleeve 42 is inserted into the In the accommodating space 33, the first sorter 40 can be positioned.

Referring to Fig. 33 and Fig. 16, the sperm sorting device 24' of the eleventh embodiment of the present invention and the sperm sorting device 24 also include the base 3 and the sorting unit 7, the difference between the two is that the sperm sorting device 24 is to position the second sorter 50 by the second protruding ring 53 on the top surface 322 of the surrounding wall 32, and the second sleeve 51 of the sperm sorting device 24' omits the second protruding ring 53. Ring 53, because the accommodating space 33 is a frusto-conical space with a wide top and a narrow bottom, and the second sleeve 52 is a frusto-conical sleeve with a wide top and a narrow bottom, the second sleeve 52 is placed Into the accommodating space 33, the second sorter 50 can be positioned.

Referring to Figure 34, the total motility and forward motility of the motile sperm 110 obtained in the present invention are compared with the sperm obtained from the semen sample stock solution and the sperm obtained from the previous device, and the obtained experimental results are in accordance with the present invention. The obtained motile sperm 110 obviously have better total motility and forward motility. Figure 34 Motile sperm (%)=number of motile sperm/total sperm number in the statistical chart, forward motile sperm (%)=number of forward motile sperm/total sperm number. The t test was used, where p<0.05 was considered significant, the symbol * indicated p<0.05, the symbol *** indicated p<0.001, and the symbol **** indicated p<0.0001.

Referring to Fig. 35 to Fig. 38, the sperm sorting device 201 of the twelfth embodiment of the present invention includes a base 3', the sorting unit 4 and a buckle unit 130.

The base 3' includes a bottom wall 31', a surrounding wall 32' extending from the periphery of the bottom wall 31 toward the top and surrounding an axis I, and a center portion of the bottom wall 31' along the axis I toward the top. Extended and tapered central projection 36 . The bottom wall 31' cooperates with the surrounding wall 32' to define an accommodating space 33' having an opening 331', and the surrounding wall 32' has an inner wall surface 321' and a top surface 322'. The central protrusion 36 is a truncated conical block with a narrow top and a wide bottom, and has a truncated conical surface 361 and several spacer ribs 362 equidistantly disposed on the truncated conical surface 361 .

The sorting unit 4 also includes the first sorter 40 , and the first sorter 40 also has the first sleeve 41 and the first permeable membrane 44 .

The buckle unit 130 is disposed between the first protruding ring 43 of the first sleeve 41 and the top surface 322' of the surrounding wall 32', and includes two clamping blocks 131 disposed on the top surface 322', and two A corresponding locking block 131 is arranged in the locking groove 132 of the first protruding ring 43, and the locking unit 130 can prevent the first separator 40 from rotating relative to the base 3', thereby improving the first The positioning effect and operation convenience of the sorter 40. It is worth noting that the card slots 132 can also be provided on the top surface 322', and the card blocks 131 can be provided on the first protruding ring 43, which have the same use effect.

The sperm sorting device 201 can drop the semen sample 100 between every two spacer ribs 362 by setting the central protrusion 36 and the spacer ribs 362 on the bottom wall 31' of the base 3'. On the truncated conical surface 361, the semen sample 100 is evenly coated on the truncated conical surface 361, thereby improving the uniformity of the semen sample 100 in the accommodating space 33'. Set operation, and position the first sorter 40 on the base 3', through the frusto-conical surface 361, the air bubbles originally located on the surface of the semen sample 100 can be concentrated on the top of the central protrusion 36 , which in turn can be easily discharged through the first permeable membrane 44 . In addition, when the first sorting liquid 120 and the motile sperm 110 are sucked, the equal interval ribs 362 of the central protrusion 36 can prevent the semen sample 100 from flowing smoothly, so as to avoid the semen Impurities in the sample 100 are sucked through the first permeable membrane 44, and the impurities can also be blocked at the junction of the frusto-conical surface 361 and the equally spaced ribs 362, so as to prevent the impurities from passing through the first permeable membrane 44. The membrane 44 is sucked, thereby improving the sorting quality.

Referring to FIG. 39 to FIG. 41 , the sperm sorting device 202 of the thirteenth embodiment of the present invention includes the base 3 ′, the sorting unit 4 , the buckle unit 130 and a liquid extractor auxiliary frame 140 .

The difference between the sperm sorting device 202 and the sperm sorting device 201 (see Fig. 35 to Fig. 38 ) is that the sperm sorting device 202 is additionally equipped with the liquid extractor auxiliary rack 140, and the liquid extractor auxiliary rack 140 includes a With the top placement ring 141 placed on the first protruding ring 43 of the first sleeve 41, several pieces of the first sleeve 42 extend from the top placement ring 141 toward the bottom end and gradually approach the axis I and are located in the first sleeve 42. The connecting ribs 142 in the first inner cylindrical surface 421 , and a bottom liquid-taking block 143 connected to the bottom ends of the connecting ribs 142 and located at the top of the first permeable membrane 44 .

It is worth noting that, the liquid extractor auxiliary frame 140 and the first set 41 can also be an integral structure in addition to the above two components. In this way, the liquid extractor auxiliary frame 140 only needs to include the The connecting ribs 142 in the inner cylindrical surface 421 and gradually approaching the axis I, and the bottom liquid-taking block 143 connected to the bottom ends of the connecting ribs 142 and located at the top of the first permeable membrane 44 are sufficient.

When in use, the needle of a liquid extractor (not shown, such as a syringe) is pressed against the bottom liquid extraction block 143, and the first sorting liquid 120 and the motile sperm 110 can be successfully sucked out, and Inadvertently puncturing the first permeable membrane 44 during operation can be avoided.

42 to 44, the sperm sorting device 203 of the fourteenth embodiment of the present invention includes the base 3' and a sorting unit 150.

The sorting unit 150 includes a third sorter 151 having a third sleeve 152 and a third permeable membrane 158 .

The third sleeve 152 has a third sleeve 153 around the axis I for being inserted into the accommodating space 33 ′, and a top end of the third sleeve 153 extends away from the axis I for positioning. The third protruding ring 154 on the top surface 322' of the surrounding wall 32', a suction ring 155 extending from the bottom end of the third sleeve 153 towards the direction close to the axis I, and a suction ring 155 disposed on the top surface 322' of the surrounding wall 32'. And the stopper slot 156 with an open top, and a pair of indicator lines 157 that are disposed on a third inner cylindrical surface 1531 of the third sleeve 153 .

The third sleeve 153 has the third inner cylindrical surface 1531, a third outer cylindrical surface 1532 surrounding the third inner cylindrical surface 1531, and a third inner cylindrical surface 1531 connected to the third outer cylindrical surface 1532. The bottom surface 1533 of the third cylinder.

The suction ring 155 has a ring top surface 1551, a ring bottom surface 1552, and a central hole 1553 connecting the ring top surface 1551 and the ring bottom surface 1552, and the ring bottom surface 1552 is formed from the third cylinder bottom surface 1533 of the third sleeve 153 The suction ring 155 gradually decreases in height from the bottom end of the third sleeve 153 to the axis I while extending towards the direction close to the axis I and gradually approaching the ring top surface 1551 .

The stopper groove 156 extends from the ring top surface 1551 toward the ring bottom surface 1552, and has a groove bottom 1561, and a communication portion 1562 connecting the groove bottom 1561 and the third permeable membrane 158. The width of the liquid limiting groove 155 is between 4 μm and 5 mm.

The third permeable membrane 158 is fixed on the third tube bottom surface 1533 of the third sleeve 153, and cooperates with the suction ring 155 and the third inner tube surface 1531 to define a third sorting chamber 159 with an open top. , the third permeable membrane 158 has a plurality of third micropores 1581 for the motile sperm 110 to pass through, a fixing portion 1582 fixed to the bottom surface 1533 of the third sleeve 153 around the axis I, and A penetrating portion 1583 extending from the fixing portion 1582 toward the axis I, the penetrating portion 1583 is flat.

When the third sleeve 153 of the third sleeve 152 is put into the accommodating space 33', the third transparent membrane 158 cooperates with the bottom wall 31' and the inner wall surface 321' of the surrounding wall 32' to define a A third sample chamber 2031 for accommodating the semen sample 100 .

The sperm sorting device 203 uses the suction ring 155 and the liquid extractor limiting groove 156, the operator only needs to insert the needle of a liquid extractor (not shown, such as a syringe) into the liquid extractor limiting groove 156 and lean against the bottom 1561 of the tank, the first sorting liquid 120 and the motile sperm 110 with strong motility can be taken out, thereby preventing the needle of the liquid extractor from piercing the third through hole. membrane 158, and because the liquid taker limiting groove 156 communicates with the third permeable membrane 158 through the communicating portion 1562, it can suck out the gap between the third permeable membrane 158 and the ring bottom surface 1552 of the suction ring 155. The first sorting liquid 120 in between can reduce the residual amount of the first sorting liquid 120 . The indicator line 157 can guide the operator to insert the needle of the liquid dispenser into the limit groove 156 of the liquid dispenser.

In addition, since the ring bottom surface 1552 extends from the third cylinder bottom surface 1533 of the third sleeve 153 toward the direction close to the axis I while gradually approaching the ring top surface 1551, the distance between the third cylinder bottom surface 1533 and the third cylinder bottom surface 1533 can be reduced. The fixed area of the fixed portion 1582 of the permeable membrane 158 can increase the permeable area of the third permeable membrane 158 relatively.

Referring to Fig. 45 to Fig. 47, the sperm sorting device 204 of the fifteenth embodiment of the present invention includes the base 3' and the sorting unit 150.

The difference between the sperm sorting device 204 and the sperm sorting device 203 (see FIG. 42 to FIG. 44 ) is that the liquid taker limiting groove 156 ′ of the third set 152 of the sperm sorting device 204 is separated from the sperm sorting device 204 . The ring top surface 1551 extends toward the ring bottom surface 1552, and has a groove bottom 1561', and a communication part 1562' connecting the groove bottom 1561' and the central hole 1553, so that the liquid extractor limiting groove 156' communicates The third permeable membrane 158, thereby, also possesses the ability to prevent the needle of the liquid extractor (not shown) from piercing the third permeable membrane 158, and can suck out the liquid located between the third permeable membrane 158 and the third permeable membrane 158. The first sorting liquid 120 between the bottom surfaces 1552 of the ring 155 is sucked up, thereby reducing the residual amount of the first sorting liquid 120 .

In addition, the suction ring 155 also has several exhaust holes 1554 connecting the ring top surface 1551 and the ring bottom surface 1552, whereby when the operator drops the first sorting liquid 120 on the third permeable membrane 158, if air bubbles are generated between the ring bottom surface 1552 and the third permeable membrane 158, the air bubbles can be discharged through the vent holes 1554 to ensure that the third permeable membrane 158 is covered with the first permeable membrane 158. The sorting liquid 120 enables the upstream motile sperm 110 to pass through the third permeable membrane 158 smoothly.

Referring to Fig. 48 to Fig. 50, the sperm sorting device 205 of the sixteenth embodiment of the present invention includes the base 3' and the sorting unit 150.

The difference between the sperm sorting device 205 and the sperm sorting device 203 (see FIG. 42 to FIG. 44 ) is that the suction ring 155 of the third set 152 of the sperm sorting device 205 is not provided with the stopper groove 156, the third transparent membrane 158' has a plurality of third micropores 1581', a fixed part 1582', and a transparent part 1583' extending from the fixed part 1582' toward the axis I and protruding toward the top end .

When in use, the operator only needs to put the needle of a liquid extractor (not shown, such as a syringe) against the ring top surface 1551 of the suction ring 155, and the first sorting liquid 120 and the active force can be taken out. The strong motile sperm 110 has the effect of preventing the needle of the liquid extractor from piercing the third permeable membrane 158 .

The sperm sorting device 205 protrudes toward the top end through the transparent portion 1583 ′, which has the effect of increasing the transparent area and reducing the space between the transparent portion 1583 ′ and the ring bottom surface 1552 of the suction ring 155 , thereby reducing the residual amount of the first sorting liquid 120 below the bottom surface 1552 of the suction ring 155 .

51 to 53, the sperm sorting device 206 of the seventeenth embodiment of the present invention includes the base 3' and a sorting unit 160.

The sorting unit 160 includes the first sorter 40 and a fourth sorter 170, the first sorter 40 also has the first sleeve 41 and the first permeable membrane 44, and the first through The permeable membrane 44 cooperates with the bottom wall 31' and the inner wall surface 321' of the surrounding wall 32' to define the first specimen chamber 211' for accommodating the semen specimen 100. The first permeable membrane 44 cooperates with the first inner cylindrical surface 421 of the first sleeve 42 to define the first sorting chamber 45 with an open top.

In addition, the first sleeve 41 also has a suction ring 48 extending from the bottom end of the first sleeve 42 towards the direction close to the axis I. The suction ring 48 has a ring top surface 481, a ring bottom surface 482, a ring connected to the The central hole 483 of the ring top surface 481 and the ring bottom surface 482, and several exhaust holes 484 communicating with the ring top surface 481 and the ring bottom surface 482, and the ring bottom surface 482 is formed from the first tube of the first sleeve 42 The bottom surface 423 extends toward the axis I and gradually approaches the ring top surface 481 , so that the height of the suction ring 48 gradually decreases from the bottom end of the first sleeve 42 to the axis I.

The fourth separator 170 has a fourth set 171 and a fourth permeable membrane 176 .

The fourth sleeve 171 has a fourth sleeve 172 around the axis I for being inserted into the first sleeve 42, and a fourth sleeve 172 extending from the top end of the fourth sleeve 172 in a direction away from the axis I for positioning The fourth protruding ring 173 on the first protruding ring 43, a suction ring 174 extending from the bottom end of the fourth sleeve 172 toward the direction close to the axis I and forming a central hole 1741, and a suction ring 174 disposed on the first protruding ring 43 The top end of the suction ring 174 is open and communicates with the liquid extractor limiting groove 175 of the central hole 1741 .

The fourth permeable membrane 176 is fixed on the bottom surface of the fourth sleeve 172, and cooperates with the suction ring 174 and a fourth inner cylindrical surface 1721 of the fourth sleeve 172 to define a fourth opening with an open top. Sorting chamber 177, and the fourth permeable membrane 176 communicates with the central hole 1741 and the liquid extractor limit groove 175, the fourth permeable membrane 176 has a passage for the motile sperm 111 with stronger motility A plurality of fourth microholes 1761 .

The sperm sorting device 206 is used to perform two-layer sperm sorting operations, and its use is similar to that of the sperm sorting device 203 (see FIGS. The needle head of the syringe) is inserted into the stopper groove 175 of the liquid taker, and the second sorting liquid 121 and the motile sperm 111 with stronger motility can be taken out directly from the stopper groove 175 of the liquid taker. Prevent the needle of the liquid extractor from piercing the fourth permeable membrane 176 .

In addition, by setting the suction ring 48 and the exhaust holes 484 on the first set 41, when the operator drops the first sorting liquid 120 on the first permeable membrane 44, if the Bubbles are generated between the ring bottom surface 482 and the first permeable membrane 44, and the bubbles can be discharged through the vent holes 484 to ensure that the first permeable membrane 44 is covered with the first sorting liquid 120, so that the upstream The motile sperm 110 can pass through the first permeable membrane 44 smoothly.

When the ring bottom surface of the suction ring 174 of the fourth sleeve 171 extends from the bottom surface of the fourth sleeve 172 toward the direction close to the axis I while gradually approaching the top surface of the ring, the suction ring 174 can also have several communication channels. The exhaust holes on the top surface and the bottom surface of the ring have the same effects as those produced by the exhaust holes 484, and will not be repeated here.

It is worth noting that: (1) the first micropores 441 of the first permeable membrane 44 of the present invention, the second micropores 551 of the second permeable membrane 55, the third permeable membrane 158 The third micropores 1581 and the fourth micropores 1761 of the fourth permeable membrane 176 can be general pores or holes formed by weaving. In addition, the first permeable membrane 44, the The second transparent film 55 , the third transparent film 158 , and the fourth transparent film 176 may have a multi-layer film stack structure. (2) In addition to being applicable to human sperm sorting, the present invention is also applicable to sperm sorting of various animals, and the first sample chamber 211, 211' is connected to the second sample chamber 241 and the third sample chamber. The height of the sample chamber 2031 can be changed accordingly, for example: (1) for sperm sorting of rams, the minimum height of these sample chambers is 67.2 μm and the highest is 35.64 cm; For the sperm sorting of goats, the minimum height of these specimen chambers is 59.39 μm and the maximum height is 16.00 cm; 13.68cm, (4) For the sperm sorting of boars, the minimum height of these specimen chambers is 54.6μm, the highest is 8.06cm, (5) For the sperm sorting of roosters, the height of these specimen chambers The lowest height is 99.36μm, and the highest is 7.65cm.

Through the above description, effects and advantages of the present invention can be summarized as follows:

1. In the present invention, by putting the first sleeve 42 of the first sorter 40 into the accommodating space 33 and positioning it, the gas originally located in the first sample chamber 211 can be squeezed out of the second out of a sample chamber 211, or squeezed into the first transparent space 212, so that the motile sperm 110 in the semen sample 100 can naturally move upwards through the first transparent space. The membrane 44 can improve the quality of sperm sorting. According to the experimental results, the motile sperm 110 obtained by the present invention have excellent total motility and forward motility.

2. The present invention can adjust the volume of the first sample chamber 211 by setting the height adjustment unit 9, 9' between the base 3 and the first sorter 40, and can also adjust the first The gap size of the through gap 212 .

3. In the present invention, the spacer 95 can be installed between the top surface 322 of the base 3 and the ring bottom surface 432 of the first convex ring 43 of the first sorter 40 to increase the size of the first sample chamber The volume of 211 can also increase the clearance of the first transparent space 212 .

4. In the present invention, by stacking the first sorter 40 or the second sorter 50 on the base 3, it can not only improve the sperm sorting quality, simplify the sperm sorting process, but also reduce the Operator error rate.

5. The present invention can use a dropper (not shown) to drop the semen sample 100 from the opening 331 of the base 3 into the accommodating space 33 without using a syringe, and the operation is more convenient.

6. In the present invention, after the sorting operation is completed, the first sorter 40 can be taken out from the base 3, and the first sorter 40 can be directly placed on the microscope stage (not shown) for all The motile sperm 110 is analyzed, and the operation is more convenient.

Seven, the present invention can take out the second sorter 50 from the first sorter 40 after the sorting operation is completed, and directly place the second sorter 50 on the stage of the microscope (not shown in the figure). ) to analyze the motile sperm 111, which is more convenient to operate.

8. In the present invention, by setting the suction ring 155 on the third set 152 of the third sorter 151, the operator only needs to press the needle of the liquid collector against the top surface 1551 of the ring to take out the suction ring 155. The first sorting liquid 120 and the motile sperm 110 have the effect of not piercing the third permeable membrane 158 .

But the above-mentioned ones are only embodiments of the present invention, and should not limit the scope of the present invention. All simple equivalent changes and modifications made according to the patent scope of the present invention and the content of the patent specification are still within the scope of the present invention. Within the scope covered by the patent of the present invention.

21, 22, 23, 24, 25, 26, 27, 28, 29, 21’, 24’, 201, 202, 203, 204, 205, 206: sperm sorting device 211,211': The first specimen chamber 212: The first transparent gap 232: The second transparent gap 241: The second sample chamber 242: The third transparent gap 3,3': base 31,31': bottom wall 32,32': around the wall 321,321': inner wall surface 322,322': top surface 323: overflow tank 324: Outer wall 33,33': accommodation space 331,331': opening 34: indicating scale 35: ring groove 36: Center bump 361: frusto-conical surface 362: spacer rib 4,4’,4”,7,7’: sorting unit 40: First sorter 41: First kit 42: First sleeve 421: the first inner cylinder surface 422: The first outer cylinder surface 423: Bottom surface of the first cylinder 43: The first convex ring 431: ring top surface 432: ring bottom 433: Circumferential surface 434: The first escape tank 44: The first permeable membrane 441: the first microhole 442: fixed part 443: Transparent part 45: The first sorting room 46: Liquid taker socket 461: jack 47: The first ring groove 48:Drain ring 481: ring top surface 482: ring bottom 483: Center hole 484: exhaust hole 50:Second sorter 51: Second Kit 52:Second sleeve 521: the second inner cylinder surface 522: Second outer cylinder surface 523: The bottom surface of the second cylinder 53: second convex ring 531: ring top surface 532: ring bottom 533: Circumferential surface 534: The second escape tank 54: inner ring 55: The second permeable membrane 551: the second microhole 56: The second sorting room 60: top cover 61: top wall 62: top around wall 63: top accommodation space 8:Height adjustment unit 81: The first stage 82: The second stage 83: The third stage 84: The first sitting part 85: The second seat 86: The third seat 9,9': height adjustment unit 91: The first internal thread segment 92: external thread segment 93: Second internal thread segment 95: Gasket 100: semen sample 110,111: motile sperm 120: the first sorting liquid 121: the second sorting fluid 130: Buckle unit 131: block 132: card slot 140: Auxiliary rack for liquid dispenser 141: top ring 142: limit rib 143: Bottom block 150: sorting unit 151: The third sorter 152: The third suite 153: The third sleeve 1531: The third inner cylinder surface 1532: The third outer cylinder surface 1533: The bottom surface of the third cylinder 154: The third convex ring 155:Drain ring 1551: ring top surface 1552: ring bottom 1553: Center hole 1554: exhaust hole 156, 156': Liquid taker limit groove 1561, 1561': the bottom of the tank 1562, 1562': Connecting part 157: indicator line 158,158': the third permeable membrane 1581,1581': the third micropore 1582, 1582': fixed part 1583,1583': Transparency 159: The third sorting room 1501: Connecting slot 2031: The third specimen chamber 160: sorting unit 170: The fourth sorter 171: The fourth kit 172: Fourth sleeve 1721: The fourth inner cylinder surface 173: The fourth convex ring 174:Drain ring 1741: Center hole 175: Limiting slot for liquid taker 176: The fourth permeable membrane 1761: The fourth micropore 177: The fourth sorting room

Other features and effects of the present invention will be clearly presented in the implementation manner with reference to the drawings, wherein: Fig. 1 is a schematic cross-sectional view of a combination of U.S. Patent No. US10,422,737B2; Fig. 2 is a three-dimensional assembled view of the first embodiment of the sperm sorting device of the present invention; Fig. 3 is a three-dimensional exploded view of the first embodiment, illustrating that the first embodiment includes a base and a sorting unit, and the sorting unit includes a first sorter; Fig. 4 is a combined sectional view of the first embodiment; Fig. 5 is a schematic diagram of the use of the first embodiment; Figure 6 is a view similar to Figure 4, illustrating that a ring groove is set at the base, and a first ring groove is set at the first sorter; Fig. 7 is a three-dimensional assembled view of the second embodiment of the sperm sorting device of the present invention; Fig. 8 is a three-dimensional exploded view of the second embodiment, illustrating that the second embodiment includes the base, a sorting unit and a top cover, and the sorting unit includes two first sorters; Fig. 9 is a combined sectional view of the second embodiment; Fig. 10 is a schematic diagram of the use of the second embodiment, illustrating that the top cover is turned upside down for placement of one of the first sorters; Fig. 11 is a three-dimensional exploded view of the third embodiment of the sperm sorting device of the present invention, illustrating that the third embodiment includes the base, a sorting unit and the top cover, and the sorting unit includes a first sorter and a second sorter; Fig. 12 is a combined sectional view of the third embodiment; Figure 13 is a schematic diagram of the use of the third embodiment; Fig. 14 is a three-dimensional assembled view of the fourth embodiment of the sperm sorting device of the present invention; Fig. 15 is a three-dimensional exploded view of the fourth embodiment, illustrating that the fourth embodiment includes the base and a sorting unit, and the sorting unit includes a second sorter; Fig. 16 is a combined sectional view of the fourth embodiment; Fig. 17 is a three-dimensional exploded view of the fifth embodiment of the sperm sorting device of the present invention, illustrating that the fifth embodiment includes the base, a sorting unit and the top cover, and the sorting unit includes two second sorting units device; Fig. 18 is a combined sectional view of the fifth embodiment; Figure 19 is a schematic view of the fifth embodiment, illustrating that the top cover is turned upside down for placement of one of the second sorters; Fig. 20 is a three-dimensional assembled view of the sixth embodiment of the sperm sorting device of the present invention, illustrating that the sixth embodiment includes the base, the sorting unit and a height adjustment unit, and the sorting unit includes a first sorting unit device, and the state where the first set is rotated 0 degrees relative to the base; Fig. 21 is a three-dimensional exploded view of the sixth embodiment; Figure 22 is a sectional view taken from Figure 20; Figure 23 is a view similar to Figure 20, illustrating the state of the first sleeve rotated 60 degrees relative to the base; Figure 24 is a sectional view taken from Figure 23; Figure 25 is a view similar to Figure 20, illustrating the state of the first sleeve rotated 120 degrees relative to the base; Figure 26 is a sectional view taken from Figure 25; Fig. 27 is a three-dimensional exploded view of the seventh embodiment of the sperm sorting device of the present invention, illustrating that the seventh embodiment includes the base, the sorting unit and a height adjustment unit, and the sorting unit includes a first sorting unit device; Fig. 28 is a combined sectional view of the seventh embodiment; Fig. 29 is a three-dimensional exploded view of the eighth embodiment of the sperm sorting device of the present invention, illustrating that the eighth embodiment includes the base, the sorting unit and a height adjustment unit, and the sorting unit includes two first points selector; Fig. 30 is a combined sectional view of the eighth embodiment; Fig. 31 is a combined sectional view of the ninth embodiment of the sperm sorting device of the present invention; Fig. 32 is a combined sectional view of the tenth embodiment of the sperm sorting device of the present invention; Fig. 33 is a combined sectional view of the eleventh embodiment of the sperm sorting device of the present invention; Figure 34 is a statistical chart showing the results of comparing the total motility and forward motility of the motile sperm obtained from the present invention with the sperm obtained from the semen sample stock solution and the sperm obtained from the previous device; Fig. 35 is a partially exploded three-dimensional view of the twelfth embodiment of the sperm sorting device of the present invention; Fig. 36 is a three-dimensional exploded view of the twelfth embodiment, illustrating that the twelfth embodiment includes a base, the sorting unit and a locking unit; Fig. 37 is an exploded perspective view of another angle of the twelfth embodiment; Fig. 38 is a combined sectional view and schematic view of the use of the twelfth embodiment; Fig. 39 is a partially exploded perspective view of the thirteenth embodiment of the sperm sorting device of the present invention; Fig. 40 is a three-dimensional exploded view of the thirteenth embodiment, illustrating that the thirteenth embodiment includes the base, the sorting unit, the buckle unit and a liquid dispenser auxiliary frame; Fig. 41 is a combined sectional view and schematic view of the use of the thirteenth embodiment; Fig. 42 is a partially exploded perspective view of the fourteenth embodiment of the sperm sorting device of the present invention; Fig. 43 is a three-dimensional exploded view of the fourteenth embodiment, illustrating that the fourteenth embodiment includes the base and a sorting unit, and the sorting unit includes a third sorter; Fig. 44 is a combined sectional view and schematic view of the use of the fourteenth embodiment; Fig. 45 is a partial perspective exploded view of the fifteenth embodiment of the sperm sorting device of the present invention; Fig. 46 is a three-dimensional exploded view of the fifteenth embodiment, illustrating that the fifteenth embodiment includes the base and the sorting unit; Fig. 47 is a combined sectional view and schematic view of the fifteenth embodiment; Fig. 48 is a partial perspective exploded view of the sixteenth embodiment of the sperm sorting device of the present invention; Fig. 49 is a three-dimensional exploded view of the sixteenth embodiment, illustrating that the sixteenth embodiment includes the base and the sorting unit; Fig. 50 is a combined sectional view and schematic view of the use of the sixteenth embodiment; Fig. 51 is a partially exploded perspective view of the seventeenth embodiment of the sperm sorting device of the present invention; Fig. 52 is a three-dimensional exploded view of the fourteenth embodiment, illustrating that the seventeenth embodiment includes the base and a sorting unit, and the sorting unit includes a fourth sorter and a fifth sorter; and Fig. 53 is a combined sectional view and schematic view of the seventeenth embodiment.

21: Sperm sorting device

211: The first sample chamber

212: The first transparent gap

3: base

31: bottom wall

32: around the wall

321: Inner wall surface

33:Accommodating space

322: top surface

323: overflow tank

4: Sorting unit

40: First sorter

41: First kit

42: First sleeve

43: The first convex ring

434: The first escape tank

44: The first permeable membrane

45: The first sorting room

46: Liquid taker socket

100: sperm sample

110: motile sperm

120: the first sorting liquid

Claims (37)

A sperm sorting device, suitable for sorting out several motile sperm from a semen sample, the sperm sorting device includes: A base, including a bottom wall, and a surrounding wall extending from the periphery of the bottom wall toward the top and surrounding an axis, the bottom wall cooperates with the surrounding wall to define an accommodating space with an opening, and the surrounding wall has an inner wall surface and a top surface; and A sorting unit, including a first sorter for positioning on the base, the first sorter has a first sleeve and a first permeable membrane, The first sleeve has a first sleeve around the axis for being inserted into the accommodating space, the first sleeve has a first inner cylindrical surface, and a first outer sleeve surrounding the first inner cylindrical surface Cylinder surface; The first permeable membrane is fixed on the first sleeve and cooperates with the first inner cylindrical surface to define a first sorting chamber with an open top. The first permeable membrane has a plurality of cells for the motile sperm to pass through first micropore; Wherein, the first transparent membrane cooperates with the bottom wall and the inner wall surface of the surrounding wall to define a first sample chamber for accommodating the semen sample. The sperm sorting device according to claim 1, wherein the sorting unit further includes another first sorter, and the first sleeve of the other first sorter is inserted into the first sorter of the first sorter. In the first sleeve, the other first classifier is stacked with the first classifier along the axis. The sperm sorting device according to claim 1, wherein the inner wall surface of the surrounding wall cooperates with the first outer cylindrical surface of the first sleeve to define a first transparent space communicating with the first sample chamber . The sperm sorting device according to claim 1, wherein the first sleeve further has a first sleeve extending from the top end of the first sleeve in a direction away from the axis for being positioned on the top surface of the surrounding wall. convex ring. The sperm sorting device according to claim 4, wherein the inner wall surface of the surrounding wall cooperates with the first outer cylindrical surface of the first sleeve to define a first transparent space communicating with the first sample chamber , the first protruding ring has a ring top surface, a ring bottom surface and a ring peripheral surface connecting the ring top surface and the ring bottom surface, and the ring bottom surface forms a ring extending from the first outer cylinder surface to the ring peripheral surface and communicating with the ring surface. The first escape groove of the first through-space. The sperm sorting device as claimed in claim 4, wherein the first convex ring has a ring top surface, a ring bottom surface and a ring peripheral surface connecting the ring top surface and the ring bottom surface, and the sperm sorting device also includes a A height adjustment unit, the height adjustment unit includes two first steps arranged on the top surface of the base and diametrically opposed to each other, the two first steps are respectively connected and the height along the axis is lower than the two first steps The second step portion that is radially opposite to each other, the third step portion that connects the two first step portions and the two second step portions and is lower in height along the axis than the two second step portions and is diametrically opposite to each other , two first seats arranged on the bottom surface of the ring and diametrically opposed to each other, two second seats respectively connected to the two first seats and diametrically opposed to each other, and two first seats connected to the two second seats The two seat portions and the third seat portion diametrically opposite to each other, when the two first seat portions are seated on the two third step portions, and the two second seat portions are seated on the two second step portions, the two seat portions are seated on the two second step portions. Two third seats are seated on the two first steps, the sperm sorting device has a first height along the axis, the first transparent gap has a first air capacity, and the first sorting chamber Having a first volume, when the two first seats are seated on the two second steps, the two second seats are on the two first steps, and the two third seats are on the two third On the step portion, the sperm sorting device has a second height along the axis, the first transparent gap has a second air volume, and the first sorting chamber has a second volume. When the two first seats seated on the two first steps, the two second seats are located on the two third steps, the two third seats are located on the two second steps, the sperm sorting device along the axis Having a third height, the first transparent gap has a third air capacity, the third air capacity is greater than the second air capacity and greater than the first air capacity, the third volume is greater than the second volume and greater than the first volume. The sperm sorting device according to claim 4, further comprising a height adjustment unit, the height adjustment unit includes a first internal thread section located on the inner wall surface of the surrounding wall of the base, and a section located on the first sleeve The first outer cylindrical surface is used for screw locking with the outer thread section of the first inner thread section. The sperm sorting device according to claim 4, further comprising a height adjustment unit, the height adjustment unit includes a first internal thread section located on the inner wall surface of the surrounding wall of the base, and a first internal thread section located on the first sleeve An outer cylindrical surface is used for screwing the external thread section of the first internal thread section, and a second internal thread section is located on the first inner cylindrical surface of the first sleeve. The sperm sorting device as claimed in claim 4, further comprising a spacer installed between the top surface of the surrounding wall of the base and the first convex ring. The sperm sorting device as claimed in claim 3, wherein the top surface of the surrounding wall forms an overflow tank. The sperm sorting device according to claim 4, wherein the sorting unit further includes a second sorter, and the first sleeve of the first set of the first sorter also has a The cylinder surface and the first cylinder bottom surface of the first outer cylinder surface, the first permeable membrane is fixed on the first cylinder bottom surface, the second separator has a second set and a second permeable membrane, the second The sleeve has a second sleeve around the axis for being inserted into the first sleeve, and a second sleeve extending from the top end of the second sleeve in a direction away from the axis for being positioned on the first collar. Two protruding rings, and an inner ring extending from the bottom end of the second sleeve toward the direction close to the axis and tapered toward the top end, the second sleeve has a second inner cylinder surface, and a ring surrounding the first sleeve The second outer cylinder surface of the two inner cylinder surfaces, and a second cylinder bottom surface connecting the second inner cylinder surface and the second outer cylinder surface, the second permeable membrane is fixed on the second cylinder bottom surface and connected to the inner cylinder surface The rings cooperate to define a second sorting chamber with an open top, and the second permeable membrane has a plurality of second micropores. The sperm sorting device as claimed in claim 11, wherein the first inner cylindrical surface of the first sleeve cooperates with the second outer cylindrical surface of the second sleeve to define a cell that communicates with the first sorting chamber. The second transparent gap. The sperm sorting device according to claim 12, wherein the second convex ring of the second sorter has a ring top surface, a ring bottom surface and a ring peripheral surface connecting the ring top surface and the ring bottom surface, the ring The bottom surface forms a second escape groove extending from the second outer cylindrical surface to the circumferential surface and communicating with the second through gap. The sperm sorting device according to claim 3, wherein the first transparent gap is less than 3 mm. The sperm sorting device according to claim 1, wherein the base further includes an annular groove disposed on the inner wall surface of the surrounding wall, and the first sorter also has a first sleeve disposed on the first sleeve. The first ring groove on the inner cylinder surface. A sperm sorting method, comprising the following steps: (A) Prepare the sperm sorting device as described in Claim 1; (B) provide a semen sample; (C) drop the semen sample into the holding space from the opening of the base; (D) Put the first sleeve of the first sorter into the accommodating space and position it, the first permeable membrane is located above the semen sample, and the semen sample is accommodated in the first sample in the body container, and the air in the first sample container is squeezed out of the first sample container; (E) drop a first sorting liquid on the first permeable membrane; (F) Stand still for 3 minutes to 60 minutes. During the standing process, several motile sperm in the semen sample will move upward and pass through the first micropores of the first permeable membrane to enter the first sorting chamber is mixed with the first sorting fluid; and (G) taking out the first sorting liquid and the motile spermatozoa with strong motility from the first permeable membrane. A sperm sorting method, comprising the following steps: (a) Prepare the sperm sorting device as described in claim 11; (b) provide a semen sample; (c) drop the semen sample into the accommodating space from the opening of the base; (d) Put the first sleeve of the first sorter into the accommodating space and position it, the first permeable membrane is located above the semen sample, and the semen sample is accommodated in the first sample in the body container, and the air in the first sample container is squeezed out of the first sample container; (e) drop a first sorting liquid on the first permeable membrane; (f) The second sleeve of the second sorter is placed into the first sorting chamber and positioned, and the second permeable membrane is located above the first permeable membrane; (g) Drop a second sorting solution on the second permeable membrane; (h) Stand still for 3 minutes to 60 minutes. During the standing process, several highly motile sperm in the semen sample will move upwards and pass through the first micropores of the first permeable membrane to enter The first sorting chamber is mixed with the first sorting liquid, and the number of motile sperm with stronger motility on the first permeable membrane will move upwards and pass through the second microscopic cells of the second permeable membrane. wells enter the second sorting chamber and mix with the second sorting fluid; and (i) Take out the second sorting solution and the motile sperm with stronger motility from the second permeable membrane. The sperm sorting device according to claim 4, further comprising a buckle unit arranged between the top surface of the surrounding wall and the first convex ring, the buckle unit includes at least one top surface of the surrounding wall The clamping block facing one of the first protruding ring, and at least one clamping groove corresponding to at least one clamping block disposed on the top surface of the surrounding wall and the other one of the first protruding ring. The sperm sorting device as claimed in claim 1, wherein the base further includes a central projection extending from the central portion of the bottom wall along the axis toward the top end with a gradually decreasing diameter. The sperm sorting device as claimed in claim 19, wherein the central protrusion has a frusto-conical surface, and several spacer ribs are equidistantly arranged on the frusto-conical surface. The sperm sorting device according to claim 1, wherein the first sorter further has a liquid collector socket disposed on the first inner cylindrical surface of the first set and located in the first sorting chamber. The sperm sorting device as claimed in claim 21, wherein the liquid collector socket has an insertion hole, and the diameter of the insertion hole is between 4 μm and 5 mm. The sperm sorting device according to claim 1, wherein the sorting unit further includes a liquid extractor auxiliary rack, and the liquid extractor auxiliary rack includes several tubes located in the first inner cylinder surface of the first sleeve and The connecting ribs gradually approaching the axis, and the bottom liquid-taking block connecting the bottom ends of the connecting ribs and located at the top of the first permeable membrane. The sperm sorting device according to claim 1, wherein the volume of the first sample chamber is between 12ul and 400ml. The sperm sorting device according to claim 1, wherein the distance between the first permeable membrane and the bottom wall is between 50 μm and 8.5 cm. The sperm sorting device according to claim 1, wherein the area of the first permeable membrane is less than 180cm². The sperm sorting device according to claim 1, wherein the diameter of each first micropore is between 2 μm and 50 μm. The sperm sorting device as claimed in claim 1, wherein the first permeable membrane has a fixed part surrounding the axis and fixed to the first sleeve, and a transparent part extending from the fixed part toward the axis part, and the penetrating part extends unevenly. A sperm sorting device, suitable for sorting out several motile sperm from a semen sample, the sperm sorting device includes: A base, including a bottom wall, and a surrounding wall extending from the periphery of the bottom wall toward the top and surrounding an axis, the bottom wall cooperates with the surrounding wall to define an accommodating space with an opening, and the surrounding wall has an inner wall surface and a top surface; and A sorting unit, including a second sorter for positioning on the base, the second sorter has a second sleeve and a second permeable membrane, The second sleeve has a second sleeve around the axis for being inserted into the accommodating space, the second sleeve has a second inner cylindrical surface, and a second outer cylinder surrounding the second inner cylindrical surface surface, and an inner ring extending from the bottom end of the second sleeve toward the direction close to the axis and tapered toward the top end; The second permeable membrane is fixed on the second sleeve and cooperates with the inner ring to define a second sorting chamber with an open top. The second permeable membrane has a plurality of first separation chambers for the motile sperm to pass through Two micropores; Wherein, the second permeable membrane cooperates with the bottom wall and the inner wall of the surrounding wall to define a second sample chamber for accommodating the semen sample. A sperm sorting device, suitable for sorting out several motile sperm from a semen sample, the sperm sorting device includes: A base, including a bottom wall, and a surrounding wall extending from the periphery of the bottom wall toward the top and surrounding an axis, the bottom wall cooperates with the surrounding wall to define an accommodating space with an opening, and the surrounding wall has an inner wall surface and a top surface; and A sorting unit, including a third sorter for positioning on the base, the third sorter has a third sleeve and a third permeable membrane, The third sleeve has a third sleeve around the axis for being inserted into the accommodating space, the third sleeve has a third inner cylindrical surface, and a third outer cylinder surrounding the third inner cylindrical surface surface, and a suction ring extending from the bottom end of the third sleeve toward the direction close to the axis; The third permeable membrane is fixed on the third sleeve and cooperates with the suction ring and the third inner cylindrical surface to define a third sorting chamber with an open top. The third permeable membrane has a A plurality of third micropores through which sperm pass; Wherein, the third transparent membrane cooperates with the bottom wall and the inner wall of the surrounding wall to define a third sample chamber for accommodating the semen sample. The sperm sorting device according to claim 30, wherein, the third sleeve of the third set of the third sorter further has a liquid-taking device limiting groove which is arranged on the suction ring and has an open top. The sperm sorting device according to claim 31, wherein the width of the limiting groove of the liquid collector is between 4 μm and 5 mm. The sperm sorting device according to claim 30, wherein the third sleeve further has a third cylinder bottom surface connecting the third inner cylinder surface and the third outer cylinder surface, and the suction ring has a ring top surface, A bottom surface of the ring, and a central hole connecting the top surface of the ring and the bottom surface of the ring, and the bottom surface of the ring of the suction ring extends from the bottom surface of the third cylinder of the third sleeve towards the direction close to the axis and gradually approaches the top surface of the ring . The sperm sorting device according to claim 33, wherein, the third permeable membrane has a fixing part surrounding the axis and fixed on the bottom surface of the third cylinder of the third sleeve, and a A penetrating portion extending axially and raised towards the tip. The sperm sorting device according to claim 33, wherein the liquid-taking device limiting groove extends from the top surface of the suction ring toward the bottom surface of the ring, the liquid-taking device limiting groove has a groove bottom, and a communication The communication part between the groove bottom and the third permeable membrane. The sperm sorting device as claimed in claim 33, wherein the suction ring further has at least one exhaust hole connecting the top surface of the ring and the bottom surface of the ring. The sperm sorting device as claimed in item 31, wherein the third set further has a pair of indicator lines that should be provided with the stop groove of the liquid picker on the third inner cylinder surface of the third sleeve.

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