阅读说明：本技术 精子分选器与精子分选方法 (Sperm sorter and sperm sorting method ) 是由 曾繁根 潘力诚 吴仁贵 曾咏钦 王绥盛 于 2019-02-01 设计创作，主要内容包括：本发明提供一种精子分选器及精子分选方法。精子分选器包括入料槽、上游分选槽、渐扩流道、回收槽以及出料槽。上游分选槽连通于入料槽与渐扩流道之间。渐扩流道具有靠近上游分选槽的入口端与远离上游分选槽的出口端,且入口端的宽度与深度分别小于出口端的宽度与深度。回收槽连通于渐扩流道的出口端。出料槽连通于渐扩流道的入口端与出口端之间的部分。(The invention provides a sperm sorting device and a sperm sorting method. The sperm separator comprises a feeding groove, an upstream separation groove, a divergent runner, a recovery groove and a discharge groove. The upstream sorting groove is communicated between the feeding groove and the divergent runner. The divergent flow channel is provided with an inlet end close to the upstream sorting groove and an outlet end far away from the upstream sorting groove, and the width and the depth of the inlet end are respectively smaller than those of the outlet end. The recovery tank is communicated with the outlet end of the divergent flow passage. The discharge chute is communicated with the part between the inlet end and the outlet end of the divergent runner.)

1. A sperm sorter, comprising:

a feeding groove;

the upstream sorting groove is communicated with the feeding groove;

the divergent flow channel is communicated between the feeding groove and the divergent flow channel, the divergent flow channel is provided with an inlet end close to the upstream sorting groove and an outlet end far away from the upstream sorting groove, and the width and the depth of the inlet end are respectively smaller than those of the outlet end;

the recovery tank is communicated with the outlet end of the divergent flow passage; and

and the discharge chute is communicated with the part between the inlet end and the outlet end of the divergent runner.

2. The sperm sorter of claim 1 further comprising:

and the filtering structure is arranged in the feeding groove so that the semen sample is filtered by the filtering structure and then fed into the upstream sorting groove.

3. The sperm sorter of claim 1 further comprising:

the discharge flow channel is communicated between the outlet end of the divergent flow channel and the recovery tank; and

and the stop block is arranged in the discharge flow channel.

4. The sperm sorter of claim 3 wherein the stop extends from the top surface of the discharge flow channel into the discharge flow channel and a thickness of one end of the stop proximate the divergent flow channel is less than a thickness of the other end of the stop proximate the recovery tank.

5. The sperm sorter of claim 3 wherein the discharge channel comprises a plurality of microchannels arranged substantially parallel to one another and in communication with the recovery tank.

6. The sperm sorter of claim 1 wherein the divergent flow path has a forward section, a mid section and a rearward section, the forward section being closest to the inlet end, the rearward section being closest to the outlet end, the mid section being located between the forward section and the rearward section, and the discharge chute being in communication with the forward section.

7. The sperm sorter of claim 6 wherein the depths of the front section, the middle section and the rear section of the divergent flow path each increase in depth in a direction from the inlet end to the outlet end, and wherein the depth of the middle section increases in the direction more than the depths of the front section and the rear section increase in the direction.

8. The sperm sorter of claim 7 wherein the width of the intermediate and rear segments, respectively, of the divergent flow path increases in a direction from the inlet end to the outlet end.

9. The sperm sorter of claim 1 further comprising a leading flow channel communicating between the inlet end of the diverging flow channel and the upstream sorting chute.

10. A method of sorting sperm comprising:

providing a sperm separator, wherein the sperm separator comprises a feeding groove, an upstream separation groove, a divergent flow channel, a recovery groove and a discharge groove, the upstream separation groove is communicated between the feeding groove and the divergent flow channel, an inlet end and an outlet end of the divergent flow channel are respectively communicated with the upstream separation groove and the recovery groove, the depth and the width of the inlet end of the divergent flow channel are respectively smaller than the depth and the width of the outlet end of the divergent flow channel, and the discharge groove is communicated with a part of the divergent flow channel between the inlet end and the outlet end;

adding a culture solution into the sperm sorter from the upstream sorting tank, and sealing the discharge tank;

adding a semen sample into the sperm sorter from the feeding groove;

after the liquid level heights of the upstream sorting tank and the recovery tank are higher than the liquid level height of the discharge tank, opening the discharge tank to enable the high-activity sperms to flow into the discharge tank; and

and taking out the high-activity sperms from the discharge chute.

11. The sperm sorting method of claim 10, further comprising:

and taking out the living sperm from the top of the upstream sorting tank.

Technical Field

The invention relates to a sperm sorter and a sperm sorting method.

Background

Infertility has gradually become an important problem in modern society, which afflicts many families. In this regard, various artificial insemination methods have been developed, such as artificial insemination (IUI), in vitro artificial Insemination (IVF), and intracytoplasmic sperm injection (ICSI). In particular, various artificial insemination procedures have different requirements for sperm motility (motility) and sperm count. Therefore, how to precisely sort the sperm according to the motility of the sperm becomes one of the important issues in the field.

Disclosure of Invention

The invention provides a sperm sorting device and a sperm sorting method, which can sort out sperms suitable for different artificial insemination methods.

The sperm separator provided by the embodiment of the invention comprises a feeding groove, an upstream separation groove, a divergent runner, a recovery groove and a discharge groove. The upstream sorting groove is communicated between the feeding groove and the divergent runner. The divergent flow channel is provided with an inlet end close to the upstream sorting groove and an outlet end far away from the upstream sorting groove, and the width and the depth of the inlet end are respectively smaller than those of the outlet end. The recovery tank is communicated with the outlet end of the divergent flow passage. The discharge chute is communicated with the part between the inlet end and the outlet end of the divergent runner.

In some embodiments, the sperm sorter further comprises a filtration structure. The filtering structure is arranged in the feeding groove, so that the semen sample is filtered by the filtering structure and then fed into the upstream sorting groove.

In some embodiments, the sperm sorter further comprises a discharge flow channel and a stop. The discharge flow passage is communicated between the outlet end of the divergent flow passage and the recovery tank. The stop block is arranged in the discharge flow passage.

In some embodiments, the stopper extends from the top surface of the discharge flow channel into the discharge flow channel, and a thickness of one end of the stopper near the divergent flow channel is smaller than a thickness of the other end of the stopper near the recovery tank.

In some embodiments, the discharge flow channel comprises a plurality of micro flow channels. The plurality of micro channels are arranged substantially in parallel with each other and are communicated with the recovery tank.

In some embodiments, the divergent channel has a front section, a middle section, and a rear section. The anterior segment is closest to the entry end, and the back end is closest to the exit end, and the middle section is located between anterior segment and the back end, and the blown down tank communicates in the middle section.

In some embodiments, the depths of the front section, the middle section and the rear section of the divergent flow passage respectively increase in the direction from the inlet end to the outlet end, and the increase in the depth of the middle section in this direction is greater than the increase in the depths of the front section and the rear section in this direction.

In some embodiments, the width of the middle section and the width of the rear section of the divergent channel increase in the direction from the inlet end to the outlet end.

In some embodiments, the sperm sorter further comprises a leading flow channel. The front guide flow channel is communicated between the inlet end of the divergent flow channel and the upstream sorting groove.

The sperm sorting method of the embodiment of the invention comprises the following steps: providing a sperm separator, wherein the sperm separator comprises a feeding groove, an upstream separation groove, a divergent flow passage, a recovery groove and a discharge groove, the upstream separation groove is communicated between the feeding groove and the divergent flow passage, an inlet end and an outlet end of the divergent flow passage are respectively communicated with the upstream separation groove and the recovery groove, the depth and the width of the inlet end of the divergent flow passage are respectively smaller than the depth and the width of the outlet end of the divergent flow passage, and the discharge groove is communicated with a part of the divergent flow passage between the inlet end and the outlet end; adding the culture solution into a sperm separator through an upstream separation tank, and sealing a discharge tank; adding the semen sample into a sperm separator through a feeding groove; when the liquid level height of the upstream sorting tank and the recovery tank is higher than that of the discharge tank, opening the discharge tank to enable the high-activity sperms to flow into the discharge tank; and taking out the high-activity sperms from the discharge chute.

In some embodiments, the sperm sorting method further comprises: live sperm were removed from the top of the upstream sorting tank.

Based on the above, the sperm sorting device of the embodiment of the invention belongs to a passive sorting device, which can utilize the movement characteristics of the sperm to perform sorting. Specifically, the sperm sorter integrates an upstream sorting tank capable of sorting sperm by a sperm upstream (swim-up) characteristic and a divergent flow path capable of sorting sperm by a sperm upstream characteristic. Therefore, the sperm sorting device can sort out different numbers of sperms and different mobility ranges according to different artificial insemination methods. In some embodiments, the divergent channel is a three-dimensional divergent channel. That is, the divergent flow path is divergent toward the outlet end in both the horizontal direction and the vertical direction, and can have a larger capacity. Thus, the amount of semen that can be processed by the sperm sorter can be increased. Furthermore, in some embodiments, by providing a filtering structure at the inlet of the sperm sorter, impurities in the semen sample can be prevented from clogging the sperm sorter. In addition, the sperm sorting method of the embodiment of the invention can generate the liquid level difference between the discharge chute and other parts of the sperm sorter by controlling whether the discharge chute is sealed or not. Accordingly, highly motile sperm in the divergent flow path can simply be forced into the discharge chute.

In order to make the aforementioned and other features and advantages of the invention more comprehensible, embodiments accompanied with figures are described in detail below.

Drawings

Fig. 1 is an exploded perspective view of a sperm sorter according to some embodiments of the present invention.

Fig. 2 is a schematic sectional view taken along line a-a' of fig. 1.

Fig. 3 is an enlarged schematic view of the leading flow passage, the divergent flow passage, and the discharge flow passage of fig. 1.

Fig. 4 is an enlarged schematic view of the pilot channel, the divergent channel, the discharge channel and the stopper of fig. 1.

Detailed Description

Fig. 1 is an exploded perspective view of a sperm sorter 10 according to some embodiments of the present invention.

Fig. 2 is a schematic sectional view taken along line a-a' of fig. 1. Fig. 3 is an enlarged schematic view of the pilot flow passage 118, the divergent flow passage 120, and the discharge flow passage 140 of fig. 1. Fig. 4 is an enlarged schematic view of the leading flow passage 118, the diverging flow passage 120, the discharging flow passage 140, and the stopper BK of fig. 1.

Referring to fig. 1, in some embodiments, the sperm separator 10 may be formed by combining an upper substrate UP and a lower substrate BP. The material of the upper substrate UP and the material of the lower substrate BP may be polymer material, glass, metal, semiconductor material, etc., and may be the same or different from each other. In some embodiments, one or more sets of alignment structures AL may be formed on the surfaces of the upper substrate UP and the lower substrate BP facing each other. Each pair of alignment structures AL may be a pair of corresponding male and female mechanisms (i.e., a pair of male and female structures). Although fig. 1 shows 4 sets of alignment structures AL disposed at 4 corners, it is well known in the art that the positions and the number of the alignment structures AL can be adjusted according to actual requirements, and the invention is not limited thereto.

Referring to fig. 1 and 2, sperm sorter 10 includes a feed chute 100. The semen sample can be temporarily stored in the feeding tank 100 and enter the other parts of the sperm separator 10 from the feeding tank 100. In some embodiments, the feed chute 100 may be divided into an upper portion and a lower portion. The upper and lower portions are formed in the upper substrate UP and the lower substrate BP, respectively, and may be combined with each other to form the feeding groove 100. In some embodiments, the feed chute 100 is a barrel-like structure. In such embodiments, the aperture of the feed chute 100 ranges from 5mm to 15 mm. In addition, the upper portion of the feeding groove 100 may protrude from the top surface of the upper substrate UP (i.e., the surface opposite to the lower substrate BP). However, it is understood that the shape and size of the feeding groove 100 can be changed according to the actual requirement, and the invention is not limited thereto.

In some embodiments, sperm sorter 10 also includes a filter structure 102. The filtering structure 102 is disposed in the feeding tank 100 to filter the semen sample fed into the feeding tank 100. The filter structure 102 may be a membrane having a plurality of perforations T. For example, the material of the film may include SU8 dry film. In addition, the plurality of perforations T may be arranged in an array or randomly in the film. For example, the pore size of the perforations T may be in the range of 40 μm to 15 μm. In some embodiments, the filter structure 102 may be disposed between the upper substrate UP and the lower substrate BP. After the upper substrate UP and the lower substrate BP are combined, the filter structure 102 may be sandwiched between the upper portion and the lower portion of the feeding groove 100. By arranging the filtering structure 102, impurities in the semen sample can be filtered. In other words, impurities in the semen sample may remain above the filter structure 102, allowing substantially impurity-free semen to pass through the filter structure 102 and into the lower portion of the feed tank 100.

The sperm sorter 10 also includes an upstream sorting tank (wiping collecting chamber)110 in communication with the feed tank 100. In some embodiments, the semen sample may enter the upstream sorting channel 110 via channel CH1 after being filtered by the filtering structure 102. Sperm with motility (or live sperm) have an upstream (swim up) characteristic, so that semen in the upstream sorting tank 110 is actively stratified. In other words, the live sperm are collected in the upper portion of the liquid in the upstream sorting tank 110, and the dead sperm are settled in the lower portion of the liquid in the upstream sorting tank 110. In some embodiments, the motility and number of viable sperm cells collected in the upper liquid layer can meet the requirements of artificial insemination (IUI) procedures. For example, artificial insemination requires more than 20000 sperm cells. Thus, the upper portion of the fluid located in the upstream sorting tank 110 may be aspirated for use in artificial insemination. It should be noted that the mobility of the sperm is described herein in terms of the speed of movement of the sperm.

In some embodiments, the upstream sort tank 110 may be configured to be divided into an upper portion and a lower portion. The upper and lower portions are formed in the upper and lower substrates UP and BP, respectively, and may be combined with each other to form the upstream sorting groove 110. In addition, a channel CH1 may be formed in the lower substrate BP connecting between the lower portion of the feed tank 100 and the lower portion of the upstream sorting tank 110. In some embodiments, the upstream sort tank 110 is a barrel-like structure. In such embodiments, the upstream sort slot 110 has an aperture ranging from 10mm to 50 mm. However, it is well known in the art that the shape and size of the upstream sorting trough 110 can be changed according to the actual requirement, and the invention is not limited thereto. Further, in some embodiments, a plurality of scale grooves R1 may be further formed on the sidewall of the upper portion of the upstream sorting chute 110. The plurality of scale grooves R1 may extend in a horizontal direction and be arranged in a vertical direction. By providing the scale groove R1, the operator is helped to observe the liquid level in the upstream sort tank 110.

The sperm sorter 10 also includes a divergent flow path 120. The divergent channel 120 is connected to the upstream sorting tank 110, and the upstream sorting tank 110 is connected between the feeding tank 100 and the divergent channel 120. In some embodiments, the divergent channel 120 is a trench disposed on the top surface of the lower substrate BP. After the upper substrate UP and the lower substrate BP are combined, the bottom surface of the upper substrate UP may define the top surface of the trench (i.e., the divergent channel 120). Divergent flow path 120 has an inlet end EN near upstream sort tank 110 and an outlet end EX far from upstream sort tank 110. In some embodiments, the inlet end EN of the divergent flow channel 120 may be in direct communication with the upstream sort trough 110. Further, the bottom surface of the inlet end EN of the divergent channel 120 may be about 0.5mm to 5mm higher than the bottom surface of the upstream sort tank 110 (as shown in fig. 2). In this way, the upper portion of the liquid in the upstream separation tank 110 can be made to flow into the divergent channel 120. In other words, live sperm can enter the divergent flow channel 120 from the upstream sort tank 110. In addition, referring to fig. 3 and 4, the width WEN and the depth DEN of the inlet end EN of the divergent channel 120 may be smaller than the width WEX and the depth DEX of the outlet end EX, respectively. For example, the width WEN may be in the range of 0.1mm to 2mm, and the depth DEN may be in the range of 0.1mm to 2 mm. On the other hand, the width WEX may be in the range of 1.5mm to 10mm, and the depth DEX may be in the range of 1mm to 3 mm. In other words, the structure of the divergent flow passage 120 fans out (fan out) both laterally and longitudinally from the inlet end EN toward the outlet end EX. Thus, the flow rate of the liquid in divergent channel 120 may gradually decrease toward outlet end EX. The highly motile sperm have a characteristic of moving up in a retrograde direction, i.e., moving back toward the inlet end EN of the divergent channel 120. Since the divergent flow path 120 has the effect of slowing down the flow rate, it is possible to avoid the highly mobile sperm from being flushed to the outlet end EX. Thus, the high motility sperm can be collected in the first half of the divergent flow channel 120 near the inlet end EN, and the lower motility sperm can flow to the outlet end EX of the divergent flow channel 120.

Referring to fig. 3 and 4, in some embodiments, the divergent channel 120 has a front section 120a, a middle section 120b and a rear section 120 c. The front section 120a is closest to the inlet end EN, the rear section 120c is closest to the outlet end EX, and the middle section 120b extends between the front section 120a and the rear section 120 c. In some embodiments, the front section 120a extends from the inlet end EN to one side of the middle section 120b, and the rear section 120c extends from the other side of the middle section 120b to the outlet end EX. In some embodiments, the length of the front section 120a ranges from 1mm to 10mm, the length of the middle section 120b ranges from 1mm to 10mm, and the length of the rear section 120c may range from 1mm to 15 mm. In addition, the width W120a and the depth D120a of the front section 120a are slightly gradually increased toward the middle section 120b, and the initial values are equal to the width WEN and the depth DEN of the inlet end EN, respectively. The width W120b and the depth D120b of the middle segment 120b greatly increase toward the rear segment 120c, respectively. On the other hand, the width W120c and the depth D120c of the rear section 120c continuously increase in the direction toward the exit end EX, respectively, up to the width WEX and the depth DEX of the exit end EX. As such, the liquid in the divergent channel 120 can have a higher flow rate in the front section 120a and a lower flow rate in the middle section 120b and the rear section 120 c. Therefore, the liquid from the upstream sorting tank 110 can smoothly enter the divergent channel 120, and can gradually decrease in speed at the middle section 120b and the rear section 120c of the divergent channel 120. The high-motility sperm can migrate to the front section 120a and enter the discharge grooves 130a and 130b through the channels CH2 and CH3 connected to the front section 120a, respectively (see fig. 3).

In some embodiments, the depth increase of the middle section 120b is greater than the depth increase of the front section 120a and the rear section 120 c. For example, the depth increase of the front section 120a may be greater than 0.1mm and less than or equal to 1 mm. The depth increase of the middle section 120b may be 0.1mm to 2 mm. The depth increase of the rear section 120c may be 0.1mm to 1.5 mm. Furthermore, the width increase of the middle section 120b may be greater than the width increase of the front section 120a and the rear section 120 c. For example, the width increase of the middle section 120b may be 0.1mm to 10mm, and the width increase of the rear section 120c may be 0.1mm to 5 mm. In such embodiments, the flow rate of the liquid in the divergent flow passage 120 is greatly reduced from the middle section 120 b.

Referring to fig. 3 and 4, in some embodiments, the sperm sorter 10 further includes a leading runner 118. The leading channel 118 communicates between the inlet end EN of the diverging channel 120 and the upstream sort channel 110. In some embodiments, the width and depth of the leading flow channel 118 are substantially constant and equal to the width WEN and depth DEN of the inlet end EN of the diverging flow channel 120, respectively.

Referring to fig. 1 and 3, the sperm sorter 10 further includes discharge chutes 130a and 130b communicating with the divergent channel 120. In some embodiments, the discharge chutes 130a and 130b communicate with the front section 120a of the divergent channel 120 to collect the high motility sperm collected in the front section 120 a. In such embodiments, the discharge chutes 130a and 130b may communicate with the front section 120a through the channels CH2 and CH3, respectively. In some embodiments, the discharge chute 130a is closer to the inlet end EN of the divergent channel 120 than the discharge chute 130 b. Generally, more motile sperm will travel back to the area closer to the inlet end EN. Thus, in such embodiments, the motility of sperm collected by the discharge chute 130a can be slightly higher than that of sperm collected by the discharge chute 130 b. For example, the flow rate of the collected sperm collected by the discharge chute 130a in the countercurrent field can be more than 180 μm/s. In this way, the sperm collected by the discharge chute 130a can be used for intracytoplasmic sperm injection (ICSI). On the other hand, the sperm collected in the discharge chute 130b can resist the flow rate in the countercurrent field of 120 μm/s to 180 μm/s, and the number of the sperm is greater than or equal to 2000. Thus, the sperm collected by the discharge chute 130b can be used for in vitro artificial Insemination (IVF). Further, in some embodiments, the discharge chutes 130a and 130b may be disposed on opposite sides of the divergent channel 120. It should be noted that although two discharge chutes are illustrated herein, the number of discharge chutes can be adjusted by one of ordinary skill in the art according to actual requirements, and the present invention is not limited thereto.

Referring to fig. 3 and 4, in some embodiments, the sperm sorter 10 further includes a discharge channel 140 and a block BK. The discharge flow path 140 communicates with the outlet end EX of the divergent flow path 120. In some embodiments, the drain channel 140 is a trench disposed on the top surface of the lower substrate BP. The width and depth of the trench may be substantially equal to the width WEX and depth DEX of the outlet end EX of the divergent channel 120. On the other hand, the block BK may be a convex structure of the lower surface of the upper substrate UP. After the upper substrate UP and the lower substrate BP are coupled to each other, the bottom surface of the block BK may define the top surface of the discharge flow channel 140. From another perspective, the block BK may also be regarded as a structure extending from the top surface of the discharge flow channel 140 toward the inside of the discharge flow channel 140. In some embodiments, the block BK can be divided into a first portion BK-1 closer to the divergent flow path 120 and a second portion BK-2 further from the divergent flow path 120. The thickness of the first portion BK-1 increases in a direction away from the diverging flow passage 120. On the other hand, the thickness of the second portion BK-2 is substantially constant and equal to the thickness of the first portion BK-1 on the side opposite the diverging flow path 120. In such embodiments, the thickness of one end of the block BK near the divergent flow passage 120 is smaller than that of the other end far from the divergent flow passage 120. In addition, dead sperm or sperm of relatively low motility can be expelled through the underside of the block BK.

In some embodiments, the discharge flow channel 140 includes a plurality of flow-blocking system microchannels 140a arranged substantially parallel to one another. Dead sperm or sperm of relatively low motility can be expelled through the fluidic channels 140a of such flow-blocking systems. In some embodiments, the flow blocking system microchannels 140a may be located below the second portion BK-2 of the block BK. By providing the flow-blocking system microchannels 140a, the flow velocity of the liquid in the divergent channel 120 can be further reduced. In other words, the sperm cell having sufficient motility can be further assisted to swim back. In some embodiments, the spacing between adjacent flow-blocking system microchannels 140a may be in the range of 0.1mm to 1 mm.

Referring to fig. 1 and 2, the sperm sorter 10 further includes a recovery tank 150. The recovery tank 150 is connected to the outlet end EX of the divergent channel 120 so that dead sperm or sperm having a relatively low motility can be discharged into the recovery tank 150. In some embodiments, the recovery tank 150 may be in communication with the outlet end EX of the divergent channel 120 via the discharge channel 140. In such embodiments, the recovery tank 150 may be in communication with a plurality of flow-blocking system microchannels 140a (shown in fig. 3). In view of the configuration of the sperm cell sorter 10, the discharge chute 130a and the discharge chute 130b may be located between the upstream sorting chute 110 and the recovery chute 150. Further, in some embodiments, the recovery tank 150 may be structured to be divided into an upper portion and a lower portion. The upper and lower portions are formed in the upper substrate UP and the lower substrate BP, respectively, and may be combined with each other to form the recovery groove 150. In some embodiments, the recovery tank 150 is a bucket-like structure. In such embodiments, the recovery tank 150 has an aperture ranging from 10mm to 80mm, and the recovery tank 150 may have an aperture size larger than the aperture of the upstream sort tank 110. However, it is well known in the art that the shape and size of the recycling tank 150 can be changed according to the actual requirement, and the invention is not limited thereto. In addition, in some embodiments, a plurality of scale grooves R2 may be further formed on the sidewall of the upper portion of the recovery tank 150. The plurality of scale grooves R2 may extend in a horizontal direction and be arranged in a vertical direction. By providing the scale groove R2, the operator can be helped to observe the liquid level in the recovery tank 150.

Based on the above, the sperm sorting device 10 according to the embodiment of the present invention is a passive sorting apparatus that can perform sorting by utilizing the movement characteristics of sperm. Specifically, the sperm sorter 10 incorporates an upstream sorting tank 110 capable of sorting sperm by a sperm upstream (swim-up) characteristic and a divergent channel 120 capable of sorting sperm by a sperm upstream characteristic. Thus, the sperm sorter 10 can sort out different numbers of sperm cells and ranges of motility for different artificial insemination procedures. In some embodiments, the diverging flow passage 120 is a three-dimensional diverging flow passage. That is, the divergent channel 120 is divergent toward the outlet end in both the horizontal direction and the vertical direction, and can have a larger capacity. In this way, the amount of semen that can be processed by the sperm sorter 10 at a time can be increased. Furthermore, in some embodiments, by providing a filter structure 102 at the inlet of the sperm sorter 10, impurities in the semen sample can be prevented from clogging the sperm sorter 10.

Next, a sperm sorting method according to some embodiments of the present invention will be described with reference to fig. 1 to 4.

First, a sperm sorter 10 as shown in fig. 1 to 4 is provided. Subsequently, the culture solution is fed to the upstream sorting tank 110. In some embodiments, the culture medium may include Phosphate Buffered Saline (PBS), F10(Ham's F-10), or the like. The culture fluid may flow into various portions of the sperm sorter 10 for wetting. In some embodiments, sufficient broth can be added such that the level of broth in feed tank 100 substantially contacts filtration structure 102. Subsequently, the discharge chutes 130a and 130b may be taped.

The semen sample is then fed into the sperm sorter 10 from the feed chute 100. After being filtered by the filtering structure 102, the semen sample can enter the upstream sorting tank 110, the divergent channel 120, the discharge channel 140 and the recovery tank 150 of the sperm sorter 10 in sequence. Since the openings of the discharge chutes 130a and 130b are sealed, the sperm can be prevented from entering the discharge chutes 130a and 130b through the divergent channel 120. As such, the liquid levels in the discharge chute 130a and the discharge chute 130b can now be lower than the liquid levels in other portions of the sperm sorter 10 (e.g., the upstream sorting chute 110 and the recovery chute 150).

When the liquid level of the discharge chute 130a and the discharge chute 130b is lower than the liquid level in the other part of the sperm sorter 10, the adhesive tape is removed, and either the discharge chute 130a or the discharge chute 130b is opened. For example, the tape may be removed when the liquid level of the upstream sort trough 110 is higher than the liquid level of the discharge trough 130 (e.g., about 0.5mm higher). Based on the difference in liquid level, highly motile sperm located in the divergent flow passage 120 (e.g., located in the forward section 120a of the divergent flow passage 120) can flow into either the discharge chute 130a or the discharge chute 130 b. In some embodiments, the amount of sperm collected can be controlled by adjusting the opening time of the discharge chute. Then, the liquid in the discharging grooves 130a and 130b can be taken out by a tool such as a pipette (pipette). From the above, the liquid in the discharging grooves 130a and 130b should include the culture solution and the sperm with high motility. In some embodiments, referring to fig. 3, the discharge chute 130a is closer to the inlet end EN of the divergent flow passage 120 than the discharge chute 130b, and the motility of the collected sperm can be slightly higher than that of the sperm collected by the discharge chute 130 b. In such embodiments, the motility and number of sperm collected by the discharge chute 130a can meet the criteria for cytoplasmic sperm injection. On the other hand, the motility and quantity of the collected sperms in the discharge chute 130b can meet the standard of in vitro artificial insemination.

In addition, the living sperm may be taken out from the upper portion of the liquid in the upstream sorting tank 110 by a pipette (pipette). In some embodiments, the motility and number of viable sperm in the upper portion of the liquid in the upstream sort tank 110 can be compatible with artificial insemination requirements.

In summary, the sperm sorting device of the present invention is a passive sorting apparatus, which can utilize the movement characteristics of the sperm to perform sorting. Specifically, the sperm sorter integrates an upstream sorting tank capable of sorting sperm by a sperm upstream (swim-up) characteristic and a divergent flow path capable of sorting sperm by a sperm upstream characteristic. Therefore, the sperm sorting device can sort out different numbers of sperms and different mobility ranges according to different artificial insemination methods. In some embodiments, the divergent channel is a three-dimensional divergent channel. That is, the divergent flow path is divergent toward the outlet end in both the horizontal direction and the vertical direction, and can have a larger capacity. Thus, the amount of semen that can be processed by the sperm sorter can be increased. Furthermore, in some embodiments, by providing a filtering structure at the inlet of the sperm sorter, impurities in the semen sample can be prevented from clogging the sperm sorter. In addition, the sperm sorting method of the embodiment of the invention can generate the liquid level difference between the discharge chute and other parts of the sperm sorter by controlling whether the discharge chute is sealed or not. Accordingly, highly motile sperm in the divergent flow path can simply be forced into the discharge chute.

Although the present invention has been described with reference to the above embodiments, it should be understood that the invention is not limited to the embodiments, but rather, may be embodied in many different forms and varied within the spirit and scope of the invention.